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  • 101
    ISSN: 1432-2072
    Keywords: Operant delayed matching task ; MK-801 ; CGS 19755 ; Scopolamine ; 8-OH-DPAT ; Cognition ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The effects of the muscarinic antagonists scopolamine HBr and MeBr, the 5-HT1A agonst 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT), and theN-methyl-d-aspartate (NMDA) antagonists MK-801 and CGS-19755 on performance of rats in a delayed matching-to-position task were examined. Pretreatment with scopolamine HBr (0.05 and 0.1 mg/kg), resulted in a delay-dependent decrease in the percentage of correct responses and discriminability (logd), but had no effect on either the latency to complete trials, or the rate of trial completion during the fixed duration session. Scopolamine MeBr (0.1 mg/kg) did not impair percent correct or increase the response latency but did decrease the rate of trial completion. 8-OH-DPAT (up to 0.3 mg/kg), had no effect on percent correct, but did induce a small decrease in discriminability. The impairment in discriminability occurred only at a dose that substantially reduced the rate of trial completion. Both MK-801 (0.05 mg/kg) and CGS 19755 (10 mg/kg) induced a delay-independent impairment in percent correct, discriminability and a reduction in the rate of trial completion without affecting latency. A lower dose of CGS 19755 (5.0 mg/kg) induced a slight impairment in discriminability without significantly affecting the other measures. Taken together, these results demonstrate some dissocation between drug-induced cognitive and motor/motivational deficits in the DMTP test. However, the data question the specificity of putative cognitive impairments reported in many previous studies with the 5-HT1A agonist 8-OH-DPAT.
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  • 102
    ISSN: 1432-2072
    Keywords: Social play ; Opioid ; Morphine Environment ; Social isolation ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract To clarify the influence of opioids on social play, the effects of morphine on playful and non-playful social behavior in juvenile rats was investigated under different conditions. Environmental variables employed were different (dim and intense) levels of illumination during testing, familiarity to the test cage, and different periods of social isolation prior to testing. Under dim light conditions, morphine markedly increased playful social behavior, such as pinning, boxing/wrestling and following/chasing, whereas non-playful social behavior such as social exploration and contact behavior was hardly affected. This effect of morphine was independent of duration of previous isolation and dose-dependent, with a maximal effect at 1.0 mg/kg. The mechanism of this effect is interpreted as an action on the rewarding aspects of play. A dose of 0.1 mg/kg of morphine abolished the initial suppression of play induced by unfamiliarity to the test cage, without influencing total levels of play. This may be an effect of morphine on the integration of sensory stimuli. Under intense light conditions, where playful behavior was completely suppressed, morphine itself hardly affected such behavior, but decreased some aspects of non-playful social behavior. These results suggest that in juvenile rats playful and non-playful forms of social behavior are differentially regulated. In addition, opioid systems may be involved at different levels in the regulation of social play.
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  • 103
    ISSN: 1432-2072
    Keywords: 5-hydroxytryptamine ; 5,7-Dihydroxytryptamine ; Operant behaviour ; Timing ; Interval bisection procedure ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract This experiment examined the effect of destroying the 5-hydroxytryptaminergic (5HTergic) pathways on rats' ability to discriminate between two durations. Rats received injections of 5,7-dihydroxytryptamine into the median and dorsal raphe nuclei or sham lesions. They were trained to press lever A following a 2-s presentation of a light and lever B following an 8-s presentation of the light. For some rats, the levers were inserted into the chamber immediately after stimulus presentation (“no-poke-requirement”); for others, the levers were not inserted until a flap covering the food tray positioned midway between the levers had been depressed (“poke-requirement”). When stable performance was attained, “probe” trials were introduced in which the light was presented for intermediate durations. Logistic functions were derived relating percent choice of lever B to log stimulus duration. Under the “no-poke-requirement” condition, the bisection point (duration yielding 50% choice of lever B) was shorter in the lesioned rats than in the control rats. Under the “poke-requirement” condition, this effect of the lesion was attenuated. There was no effect of the lesion on the Weber fraction under either condition. The levels of 5HT and 5-hydroxyindoleacetic acid were reduced in the brains of the lesioned rats, but the levels of noradrenaline and dopamine were not altered. It is proposed that rats may attain accurate timing under the interval bisection task by moving from one lever to the other during stimulus presentation; this movement may be facilitated by destruction of the 5HTergic pathways. Accurate timing is still possible when this movement is suppressed by the introduction of a “poke requirement”; however, in this case timing is not affected by 5HT depletion.
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  • 104
    ISSN: 1432-2072
    Keywords: Clozapine ; Ritanserin ; Atypical neuroleptic ; Lick rhythm ; Oscillator slowing ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract In order to assess the effects of the atypical neuroleptic clozapine on orolingual competence in rats, tongue function was measured by quantitating the rhythm of tongue movements after acute (1.0, 3.0, 6.0 mg/kg) or subchronic intraperitoneal treatment (1.5, 3.0, 4.5 mg/kg, each dose for at least 7 days) with the drug. Thirsty rats were trained to lick water from a force-sensing disk by thrusting the tongue through a 12-mm-diameter hole to strike the horizontal disk located 5 mm below the hole. Number of licks in 2 min and rhythm of tongue movements (as determined by Fourier analysis of the force-time signal) were each dose dependently reduced in the acute dose-effect phase of the study. In the subchronic study number of licks exhibited tolerance, but the slowing of lick rhythm did not show tolerance. An acute dose range of the serotonin antagonist ritanserin (0.5, 1.0, 2.0, 4.0 mg/kg) was also studied in the same rats. Ritanserin had no effect on any of the measures of orolingual function. The clozapine result was replicated in a second study using younger, drug naive rats. The results for clozapine were contrasted with previous reports indicating that haloperidol has little effect on lick rhythm. Additional discussion evaluated the possible contribution of neurotransmitter receptors on motor neurons of the hypoglossal nucleus to the observed rhythm slowing induced by clozapine.
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  • 105
    ISSN: 1432-2072
    Keywords: Operant behavior ; Amphetamine ; Methamphetamine ; Methylenedioxymethamphetamine ; Fenfluramine ; Parachloroamphetamine ; Dopamine ; Serotonin ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Amphetamine and related compounds have previously been shown to differentially release dopamine (DA) and serotonin (5HT) in vivo and in vitro. The purpose of this report is directly to compare five amphetamine analogs on differential reinforcement of low rate 36-s (DRL 36-s) schedule performance, and to determine whether the reported increases in dopamine and/or serotonin release induced by these drugs can be related to observed behavioral differences. Amphetamine (AMPH) and methamphetamine (METH) induced large increases in response rate, methylene-dioxymethamphetamine (MDMA) and para-chloroamphetamine (PCA) caused small increases in response rate, while fenfluramine (FEN) had no effect on response rate. AMPH, METH, PCA and MDMA caused a dose-dependent decrease in reinforcement rate, and FEN had no effect on reinforcement rate. AMPH, METH, and PCA but not FEN, shifted the peak of the inter-response time (IRT) distribution toward shorter intervals, MDMA decreased peak location only at the highest dose. All five drugs caused a dose-dependent decrease in peak area, indicating a loss of schedule control on the DRL 36-s schedule. Consistent with in vitro and in vivo release studies, the differential results of these five drugs on DRL 36-s schedule performance suggest a predominant dopamine role for AMPH and METH, a predominant serotonin role for FEN, and different degrees of combined dopaminergic and serotonergic roles for MDMA and PCA in the mediation of the task.
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  • 106
    ISSN: 1432-2072
    Keywords: Anxiety ; Plus maze ; Rat ; Benzodiazepine receptor ; DMCM ; FG 7142 ; Yohimbine ; Pentylenetetrazol ; β-Carboline ; Inverse agonist
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The present series of experiments examined the effects of five benzodiazepine receptor (BZR) partial inverse agonists on the behaviour of rats on an elevated plus maze. The drugs were tested in a standard plus maze with 3-cm walls added to the open arms, as this has been shown to increase the sensitivity of the plus maze to anxiogenic-like drug effects (Jones and Cole 1995). The drugs tested were FG 7142 (0–100 mg/kg),β-CCE (0–30 mg/kg), ZK 132 556 (0–100 mg/kg), ZK 90 886 (0–30 mg/kg) and Ro 15–4513 (0–30 mg/kg). In addition, to allow a comparison with previous studies, the effects of three reference substances, DMCM (0–2.5 mg/kg), pentylenetetrazol (PTZ; 0–30 mg/kg) and yohimbine (0–5 mg/kg), were also examined. These three reference compounds produced a dose-dependent reduction in the duration of open arm exploration and the total number of open arm entries, indicative of anxiogenic-like effects. DMCM produced significant effects at the doses of 1.25 and 2.5 mg/kg, PTZ at 30 mg/kg, and yohimbine at 5 mg/kg. The BZR partial inverse agonist FG 7142 (10, 30 and 100 mg/kg) also reduced the duration of open arm exploration and the total number of arm entries. The minimally effective dose resulted in a receptor occupancy of approximately 80%. Ro 15–4513 also produced anxiogenic-like effects, but only at a dose (30 mg/kg) that resulted in a receptor occupancy of approximately 95%. In contrast, the other BZR partial inverse agonists, ZK 132 553 and ZK 90 886, did not significantly reduce the duration of open arm exploration, even at doses that produced greater than 95% receptor occupancies.β-CCE also did not reduce open arm exploration at any dose tested (0–30 mg/kg). The GABA shift, a biochemical index of intrinsic activity, indicates that these latter three compounds are more inverse agonistic than Ro 15–4513. In summary, these results demonstrate that not all BZR receptor partial inverse agonists have anxiogenic-like activity in the rat plus maze, and that the GABA shift, a biochemical index of intrinsic efficacy, does not predict which BZR partial inverse agonists are anxiogenic.
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  • 107
    ISSN: 1432-2072
    Keywords: Anxiety ; Cholecystokinin ; CCK-A and CCK-B receptor antagonists ; CCK-B receptor agonists ; Behavioural suppression ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The effects of cholecystokinin (CCK) receptor ligands were studied in the rat safety signal withdrawal conflict procedure, an operant paradigm sensitive to both anxiolytic and anxiogenic compounds. In this procedure, behavioural suppression of lever pressing for food was induced by the withdrawal of a conditioned signal for safety without the usual presentation of a conditioned signal for danger. The compounds tested were selective CCK-B antagonists [CI-988 (0.01–1 mg/kg SC), L-365,260 (0.004–2 mg/kg IP) and LY 262,691 (0.001–1 mg/kg SC)], CCK-B agonists [CCK-4 (0.01–1 mg/kg SC) and BC 264 (0.004–1 mg/kg IP)] and CCK-A antagonists [devazepide (0.001–1 mg/kg SC) and lorglumide (0.01–1 mg/kg SC)]. None of these drugs induced the expected behavioural effects, i.e. an anxiolytic-like release of the behavioural suppression with CCK-B and, possibly, CCK-A antagonists and/or a further reduction of lever pressing with CCK-B agonists, indicative of an anxiogenic-like potential. In contrast, the established anxiolytic lorazepam (0.06–0.25 mg/kg IP), as well as diazepam (2 mg/kg IP) and buspirone (0.25 mg/kg SC) used as positive control drugs, released the suppression of pressing for food during the period associated with the safety signal withdrawal, whereas picrotoxin (1 mg/kg IP), used as an anxiogenic control, further reduced responding during this conflict period. The present results contrast with a series of published data suggesting the involvement of CCK processes in anxiety-related behaviour in rodent models such as the elevated plus-maze or the light:dark two compartment test, and in panic disorders in humans. They indicate that the behavioural effects of one category of drugs might vary considerably, depending on the experimental situation. Furthermore, they allow the conclusion that anticipatory anxiety generated by withdrawal of conditioned signals for safety does not involve CCK-related processes.
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  • 108
    Electronic Resource
    Electronic Resource
    Springer
    Psychopharmacology 121 (1995), S. 451-460 
    ISSN: 1432-2072
    Keywords: Alcohol withdrawal ; Diazepam ; Kindling ; Seizures ; Rat ; Paradigm
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The purpose of the present experiment was to study the “kindling” hypothesis of alcohol withdrawal stating that exposure to repeated episodes of alcohol withdrawal results in an increased severity of subsequent withdrawal reactions. Two groups of male Wistar rats were subjected to 13 episodes of 2 days severe alcohol intoxication and 5 days alcohol withdrawal. Animals in the control group (n=80) developed clinical withdrawal signs following each intoxication episode. In the diazepam group (n=80) the withdrawal reactions during episodes 1–9 were blocked by intraperitoneal diazepam administration (0–30 mg/kg) 8, 11 and 15 h into withdrawal. During episode 10–13 diazepam treatment was terminated and convulsive withdrawal behaviour was observed 9–15 h after last alcohol dose. The probability of seizure activity during these four withdrawal episodes was calculated as 0.239 and 0.066 in the control and the diazepam groups, respectively. Based on Monte Carlo simulation techniques, this difference was found to be statistically significant (P〈0.05). No differences in the non-convulsive alcohol withdrawal symptoms tremor, hyperactivity and rigidity were detected between controls and diazepam animals after diazepam treatment. It was concluded that the increased convulsive behaviour in the control group was caused by cumulated kindling-like cerebral alterations during the previous repeated alcohol withdrawal phases.
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  • 109
    ISSN: 1432-2072
    Keywords: Psychostimulant ; Amphetamine ; Stress ; Long-term sensitization ; Social isolation ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The aim of the present study was to assess the influence of experimential factors on the vulnerability of rats to develop amphetamine (AMPH)- and stressor-induced behavioral sensitization. Young male Wistar rats with previous social experience were isolated from their peers for 2 weeks. 1) The effect of this short-lasting social deprivation were: a) a reduced tendency to explore a fearful environment; b) a prolonged exploratory activity in response to a novel but little fearful environment; and c) a dose-dependent increase in the psychomotor stimulation induced by systemic AMPH injection. 2) After repeated AMPH injections (injection every other day for 10 days), isolated rats exhibited behavioral sensitization at lower doses (0.5 and 0.75 mg/kg) than those required for group-housed rats (1 mg/kg). 3) After being submitted to a repeated stressor (3, 7 or 14 footshock sessions, with 2 days between sessions), the isolated rats exhibited a greater increase in the behavioral responsivity to a subsequent AMPH challenge (1 mg/kg) than did the group-housed rats regardless of the number of stress sessions. In conclusion, these results suggest that experiential factors such as privation of contact with peers (social isolation) may make rats more vulnerable to the long-term repercussions of chronic environmental and pharmacological challenges.
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  • 110
    ISSN: 1432-2072
    Keywords: Morphine ; Self-administration Saccharin ; Taste ; Opioid ; Reward ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract An experiment was performed to determine the relationship between saccharin preference and the self-administration of morphine via the oral and intravenous routes. On the basis of voluntary intake of a saccharin solution by male rats, low and high preference groups were formed. Rats selected for high saccharin preference self-administered more morphine intravenously than rats selected for low preference. The two groups did not differ in oral morphine intake. The positive relationship between the intake of saccharin and intravenous morphine self-administration may be due to their mediation by a common mechanism. Measures of taste sensitivity or preference may be useful in identifying individuals at risk for drug abuse.
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  • 111
    ISSN: 1432-2072
    Keywords: D1 dopamine agonists ; Quinpirole ; MPTP Marmoset ; Rat ; 6-Hydroxydopamine ; Behaviour
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The effects of co-administration of quinpirole with benzazepine D1 dopamine (DA) agonists possessing full/supramaximal (SKF 80723 and SKF 82958), partial (SKF 38393 and SKF 75670) and no efficacies (SKF 83959) in stimulating adenylate cyclase (AC) were investigated in rodent and primate models of Parkinson's disease (PD). In rats with a unilateral 6-hydroxydopamine (6-OHDA) lesion of the medial forebrain bundle, co-administration of SKF 38393 (7,8-dihydroxy-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine), SKF 75670 (3-CH3 analogue), SKF 80723 (6-Br analogue), SKF 83959 (6-Cl, 3-CH3, 3′-CH3 analogue) and SKF 82958 (6-Cl, 3-C3H5 analogue) strongly potentiated the contralateral circling induced by quinpirole. In MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) treated common marmosets, administration of quinpirole alone increased locomotor activity and reversed motor deficits. Grooming and oral activity were unaltered. Co-administration of SKF 38393 and SKF 75670 inhibited the quinpirole-induced changes in locomotor activity and motor disability. The combined treatment of SKF 80723 or SKF 82958 with quinpirole had no overall effect on locomotor activity or motor disability. In contrast, SKF 83959 extended the duration of the quinpirole-induced increase in locomotor activity with corresponding decreases in motor disability. Co-administration of high doses of SKF 82958 and more especially SKF 83959 and SKF 80723, with quinpirole induced hyperexcitability and seizures. Oral activity and grooming were unaltered following the co-administration of benzazepine derivatives with quinpirole. The ability of some benzazepine D1 DA agonists to prolong the antiparkinsonian effects of quinpirole in the MPTP-treated marmoset may indicate a role for certain D1 DA agonists in the clinical treatment of PD. In general, the behavioural responses to the combined administration of benzazepines with quinpirole in the 6-OHDA lesioned rat and more especially the MPTP-treated marmoset failed to correlate with their ability to stimulate AC. These observations further implicate a behavioural role for D1 DA receptors not linked to AC.
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  • 112
    ISSN: 1432-2072
    Keywords: d-Fenfluramine ; Brain kinetics ; Indole-depleting effect ; Inducers of drug metabolism ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The effects of pretreatment with inducers of hepatic cytochrome P450 isoenzymes (phenobarbital, dexamethasone andβ-naphthoflavone) on the metabolism ofd-fenfluramine (d-F) and its acute and long-lasting indole-depleting effects were studied in rats, in an effort to obtain further information on the importance of hepatic drug metabolism in relation to its neurochemical actions. Twenty-four hours after the last dose of each inducer, rats were injected withd-F hydrochloride (5 mg/kg, IP) and killed at various times thereafter for parallel determination of indoles and drug concentrations in plasma and brain. Additional rats were treated as above and killed 1 week afterd-F hydrochloride (5 and 10 mg/kg) to study the recovery of indole in the cortex, a particularly sensitive brain area. Phenobarbital andβ-naphthoflavone and, to a lesser degree, dexamethasone, stimulated the metabolism ofd-F, as evidenced by a decrease in plasma and brain areas under the curve (AUC) compared to vehicle-treated rats. This indicated that multiple isoenzymes are capable of mediating the drug's metabolism, primarily byN-dealkylation tod-norfenfluramine (d-NF). None of the inducers raised plasma and brain AUC of the nor-derivative, and in fact phenobarbital and particularlyβ-naphthoflavone reduced it. These different effects were even apparent in rats givend-NF (2.5 mg/kg), indicating that both phenobarbital andβ-naphthoflavone also stimulate the sequential metabolism of the nor-metabolite (byN-deamintaion) which, however, is apparently enhanced most actively byβ-naphthoflavone-inducible forms of P-450. Total “active” brain concentrations (d-F+d-NF) after the different pretreatments were in the order ofβ-naphthoflavone 〈 phenobarbital 〈 dexamethasone ≤ vehicle. Interestingly,β-naphthoflavone rapidly reversed the depletion of brain indoles caused byd-F (andd-NF); phenobarbital provided partial protection and dexamethasone did not appreciably modify either the acute or long-term neurochemical effects of the drug. The fact that phenobarbital affectedd-NF kinetics less thanβ-naphthoflavone, and provided only partial protection against the acute and long-lasting neurochemical effects of high doses ofd-F, further stresses the critical role ofd-NF in the neurochemical outcome of its parent drug. These findings support the view that the degree and duration of the indole-depleting effects are related to critical brain concentrations of the parent compound and its nor-derivative, and provide indirect evidence that hepatic metabolites other thand-NF are unlikely to play any role in the neurochemical effects of high doses ofd-F in rats.
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  • 113
    ISSN: 1432-2072
    Keywords: Antipsychotic drugs ; D4 dopamine receptor ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The affinities of 13 atypical and 12 typical antipsychotic drugs for the cloned rat D4 dopamine receptor and the D4/D2 ratios were examined. Of the atypical antipsychotic drugs tested, only clozapine, risperidone, olanzapine, zotepine and tiospirone had affinities less than 20 nM. In fact, many atypical antipsychotic drugs had relatively low affinities for the cloned rat D4 receptor, with Ki values greater than 100 nM (Seroquel, fluperlapine, tenilapine, FG5803 and melperone). Additionally, several typical antipsychotic drugs had high affinities for the cloned rat D4 receptor, with Kis less than 20 nM (loxapine, chlorpromazine, fluphenazine, mesoridazine, thioridazine and trifluoroperazine). The ratios of D2/D4 affinities did not differentiate between these two types of antipsychotic drugs. Thus, D4 dopamine receptor affinity, used as a single measure, does not distinguish between the group of typical and atypical antipsychotic drugs analyzed.
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  • 114
    ISSN: 1432-2072
    Keywords: Yawning ; Penile erections ; 5-HT1A agonists ; Tertatolol ; pCPA ; Apomorphine ; Physostigmine ; mCPP ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Apomorphine and mCPP induced yawning associated with penile erections in rats, whereas physostigmine induced only yawns. Apomorphineinduced yawning and penile erections were antagonized by low doses of raclopride, whereas physostigmineinduced yawning and mCPP-induced effects were only partly inhibited at high doses of raclopride. Scopolamine as well as clozapine antagonized yawning and penile erections induced by apomorphine, mCPP and physostigmine. Similarly, the 5-HT1A agonists 8-OH-DPAT and S 14506 inhibited yawning and penile erections induced by apomorphine, mCPP and physostigmine, and at similar doses induced lower lip retraction and hyperreactivity to handling. The β/5-HT1A antagonist tertatolol reversed the inhibitory effects of 8-OH-DPAT and S 14506 on drug-induced yawning and penile erections and increased apomorphine-and physostigmine-induced yawn frequency but not penile erection frequency. Like tertatolol, propranolol increased apomorphine- and physostigmine-induced yawn frequency, whereas ICI 118551 increased only physostigmine-induced yawning. 8-OH-DPAT-and S 14506-induced lower lip retraction and hyperre-activity to handling were also significantly antagonized by tertatolol. Finally,p-chlorophenylalanine pretreatment produced about 95% depletion in 5-HT in hypothalamus, hippocampus, striatum and frontal cortex and modified neither the responses of the inducing drugs nor the inhibitory effects of 8-OH-DPAT and S 14506 on drug-induced yawning and penile erections. These data suggest (i) that a final cholinergic mechanism blocked by scopolamine and clozapine is involved in drug-induced yawning and penile erections, (ii) that the effects of S 14506 depend upon stimulation of 5-HT1A receptors, (iii) that the inhibitory effects of 5-HT1A agonists on yawning and penile erections probably occurred at final steps on the pathways involved in the expression of yawning and penile erections and that these effects could be related to other effects of the 5-HT1A agonists, (iv) that a tonic inhibitory β influence could interfere with the expression of yawning but not with that of penile erections and (v) that presynaptic 5-HT1A receptors do not seem to play an important role in the inhibitory effects of 5-HT1A agonists on drug-induced yawning and penile erections in rats.
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  • 115
    ISSN: 1432-2072
    Keywords: Stretched approach posture ; Ambivalent behaviour ; Intention movements ; Ethological observation ; Rat ; Anxiety disorders ; Benzodiazepines ; 5-HT1A receptor agonists ; 5-HT uptake inhibitors ; Clonidine ; Clorgyline ; Anxiogenic drugs
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The effect of various psychotropic drugs on the ambivalent behaviour “stretched approach posture” (SAP) in the rat was assessed. SAP was elicited after a mild startle reaction due to physical contact with an electrified prod at one end of a straight runway. Using ethological observation methods, SAP as well as intention movements, prod contact, crossings, rearing, exploration, grooming and immobility were recorded. The benzodiazepine receptor agonists chlordiazepoxide, diazepam and alprazolam, the 5-HT1A receptor agonists flesinoxan and ipsapirone and the 5-HT uptake inhibitor clomipramine selectively (no effect on crossings) reduced SAP. Except for alprazolam, these drugs also reduced intention movements. In addition, chlordiazepoxide and diazepam enhanced prod contact. Reductions of SAP and intentions with concomitant reductions of crossings (nonspecific anti-ambivalent effects) were established for the α2-adrenoceptor agonist clonidine and the MAO inhibitor clorgyline. The 5-HT uptake inhibitor fluvoxamine suppressed intention movements, but not SAP. The mixed 5-HT/NA uptake inhibitor imipramine did not significantly affect SAP or intentions, but reduced crossings. The 5-HT2C/1B receptor agonist m-CPP, the inverse BZD receptor agonists FG 7142 and DMCM, and the α2-adrenoceptor antagonist yohimbine, to all of which putative anxiogenic effects have been ascribed, had no effect on SAP directed towards the prod. m-CPP, however, produced an increase in the stretched posture directed away from the prod (SAwayP). FG 7142 reduced intentions while strongly enhancing immobility (freezing). SAwayP and/or freezing may possibly reflect anxiogenic properties of drugs. The putative anxiogenic drug pentylenetetrazol false positively reduced SAP while increasing exploration. The dopamine-D2 receptor antagonist haloperidol and the catecholamine releaserdl-amphetamine had no effect on ambivalent behaviour. The muscarine receptor antagonist scopolamine reduced SAP and intentions while stimulating crossings. Finally, the 5-HT2C receptor antagonist ritanserine, the CCKA receptor antagonist devazepide, the CCKB receptor antagonist L-365.260 and the strychnine-insensitive glycine site antagonist 7-Cl-kynurenic acid were without effect on the behaviours in this paradigm using single doses. In conclusion, SAP and intention movements were reduced selectively by anxiolytic agents from different classes, including benzodiazepine receptor agonists, 5-HT1A receptor agonists and a 5-HT uptake inhibitor, whereas an α2-adrenoceptor agonist and a MAO inhibitor reduced SAP non-selectively. SAP in relation to other behaviours may therefore serve as a valuable paradigm to characterize anxiolytic drugs.
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  • 116
    Electronic Resource
    Electronic Resource
    Springer
    Psychopharmacology 121 (1995), S. 158-163 
    ISSN: 1432-2072
    Keywords: Methamphetamine ; Behavioral sensitization ; Scopolamine ; Acetylcholine ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Cholinergic neurotransmission has been implicated in various forms of neural plasticity such as kindling and learning. We have previously shown that blockade of muscarinic cholinergic receptors prevents the development of locomotor sensitization to methamphetamine. The present study was conducted to examine whether scopolamine, a muscarinic cholinergic antagonist, would also block augmentation of stereotypy induced by chronic methamphetamine (MA) treatment. Rats treated with MA (2.5 mg/kg, SC) for 10 days indicated significantly enhanced stereotyped behavior when tested with MA (2.5 mg/kg) after a 7-to 8- day withdrawal. Pretreatment with scopolamine (3 mg/kg) prior to MA administration prevented the augmentation of stereotypy. Rats treated with scopolamine alone showed no difference in MA-induced stereotypy compared to those treated with saline. Scopolamine methylbromide, a derivative of scopolamine that does not easily cross the blood-brain barrier, had no effect on the augmentation of stereotypy. These results suggest that stimulation of central muscarinic cholinergic receptors plays a role in the development of sensitization to the stereotypy stimulating effect of methamphetamine.
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  • 117
    ISSN: 1432-2072
    Keywords: Acoustic startle response ; Prepulse inhibition ; Schizophrenia ; Mesocorticolimbic DA system ; Dopamine ; Serotonin ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The effect of local injection of pertussis toxin (PTX) into the ventral tegmental area (VTA) on acoustic startle in rats was investigated. The PTX treatment caused only minor effects of its own on the acoustic startle response (ASR) or prepulse inhibition (PPI) of acoustic startle. However, systemic treatment with the indirect DA receptor agonist, amphetamine (2 mg/kg, SC) caused a significant increase in ASR magnitude and a significant disruption of PPI in PTX-treated rats while no such effects were observed in sham-treated rats. Treatment with the direct DA receptor agonist, apomorphine (2 mg/kg, SC), caused a significant disruption of PPI, an effect that was observed in both PTX-and sham-treated rats. Treatment with the 5-HT1A receptor agonist, 8-OH-DPAT (0.5 mg/kg, SC), did not affect PPI in either group but caused a marked increase in ASR magnitude in sham-treated rats. Interestingly, this effect was blocked in PTX-treated rats. The present results suggest that local injection of PTX into the VTA causes an increased sensitivity to the behavioural effects of psychostimulants on acoustic startle and may also suggest that intact midbrain 5-HT1A receptors are essential for the effect of 5-HT1A agonists on acoustic startle.
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  • 118
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    Psychopharmacology 119 (1995), S. 299-304 
    ISSN: 1432-2072
    Keywords: Dopamine ; Autoreceptor ; Sulpiride ; Quinpirole ; Locomotion ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Low doses of dopamine (DA) agonists such as the D2 receptor subfamily agonist quinpirole are thought to stimulate DA autoreceptors selectively, thereby inhibiting locomotor activity. High doses of quinpirole initially suppress and later activate locomotion during a single test session; the activation is presumably due to stimulation of postsynaptic receptors. The aim of this study was to investigate whether pretreatment with a selective DA D2 receptor antagonist, sulpiride, could block the putative autoreceptor-mediated inhibition at a lower dose than was required to block the postsynaptically mediated activation. Male and female 30-day-old rats were injected SC with one of eight doses of sulpiride (0.313–40 mg/kg) or the vehicle. Sixty minutes later, rats were injected SC with 0.2 mg/kg quinpirole or the vehicle. Five minutes after the second injection, rats were placed in automated activity monitors which recorded locomotor behavior for 60 min at 5-min intervals. Quinpirole at this dose first suppressed and later increased locomotor activity. Sulpiride pretreatment dose-dependently reversed both the early inhibition and later activation of quinpirole-induced locomotion. However, sulpiride did not block the quinpirole-induced early suppression at a lower dose than was required to block the later activation. Thus, there was no evidence that the locomotor suppression elicited by quinpirole is mediated by a more sensitive subset of DA receptors.
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  • 119
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    Psychopharmacology 119 (1995), S. 399-404 
    ISSN: 1432-2072
    Keywords: Rat ; State-dependency ; Diazepam ; FG 8205 ; Zolpidem ; Agonist ; Partial agonist ; Benzodiazepine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The state-dependent effect of the BZ-receptor agonist diazepam (1.25–10 mg/kg), the partial agonist FG 8205 (0.5–4.0 mg/kg) and the BZ1-receptor agonist zolpidem (0.25–2 mg/kg) were investigated in rats. During daily sessions, animals were trained to acquire FR10 lever pressing for food reinforcement whilst under the influence of the agonists, using an operant technique. Forty-eight hours after the final training session under drug, their performance of the FR10 was evaluated during a test session, carried out following vehicle administration only. Neither diazepam, nor FG 8205 impaired acquisition of the task. In the group treated with 2 mg/kg zolpidem, six out of eight rats failed to learn within 20 sessions, but the smaller doses were without effect on acquisition. When drug treatment was withdrawn, there was evidence that all three of the agonists tested produced state-dependency. This was apparent in the form of longer latencies to obtain reinforcement and decreased lever pressing rates. The significance of these findings are discussed in the context of the relationship between the state-dependent effects of BZ-receptor agonists and their other properties, and the receptor subtypes which might underly these effects.
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  • 120
    ISSN: 1432-2072
    Keywords: Serotonin1A agonist ; 8-OH-DPAT ; Lever pressing ; Stimulant ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract In general, the effects of 8-OH-DPAT on the body temperature of rats or in inducing the 5-HT syndrome show rapid tolerance. However, in contrast, the 8-OH-DPAT-induced increase in the activity of rats in a two-way active avoidance task only occurs after repeated administration, i.e. there is sensitisation. The present study was conducted to examine whether this developing hyperactivity may also be expressed as increased rates of lever press responding, and if so, under which conditions it occurs. Rats were trained to press levers under fixed interval 60-s (FI 60) or differential reinforcement of low rates 20-s or 72-s (DRL20, DRL72) schedules of food reinforcement. Groups of trained rats were then treated daily 5 min before testing with doses of 0.01, 0.1 and 1.0 mg/kg 8-OH-DPAT SC for 10–21 days. In all three procedures, in the first couple of days of drug treatment, 8-OH-DPAT generally suppressed lever pressing in a dose-dependent manner. Thereafter, tolerance to this effect was seen to a greater (DRL20, DRL72) or lesser (FI60) extent. Some evidence for stimulation of low rates of lever press responding was seen after 10 days treatment under FI60, but not in DRL20 or DRL72 during short 30 to 60 min long daytime tests although in the latter case, the rats responded to the stimulating effects of 0.8 mg/kg SC amphetamine administered once at the end of the experiment. However, when rats were allowed to respond under DRL72 testing for 12 h during the night, after 10 days treatment a clear stimulation of lever pressing was observed. This stimulation was not specific to lever pressing, however, since a stimulation of entries into the food tray and licking were also seen. From these results, it may be concluded that the stimulating effect of 8-OH-DPAT after repeated administration may be expressed as increased rates of lever pressing, but not under all conditions in which psychomotor stimulation by amphetamine is seen. The potential for 8-OH-DPAT and related compounds to stimulate motor responding in this way should be taken into account when interpreting the effects of these drugs in animal models of psychiatric disorders.
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  • 121
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    Psychopharmacology 118 (1995), S. 310-315 
    ISSN: 1432-2072
    Keywords: MK-801 ; APV and CGP 43487 ; Apomorphine-induced behavior ; Motor activity ; Rota-rod test ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The administration to rats of different doses of the non competitive NMDA receptor blocker MK-801 (0.03–1 mg/kg IP) induced stimulation or reduction of locomotor activity, depending on the dose, whereas the competitive NMDA antagonists CGP 43487 (0.188–6 mg/kg IP) and APV (2.5–20 μg/rat ICV) inhibited locomotion at the highest doses. Unlike MK-801 and APV treatment, the administration of CGP 43487 did not induce impairment of rota-rod test performance. Both competitive and non-competitive NMDA antagonists, at doses devoid of any behavioral effect per se, potentiated the responses elicited by apomorphine (0.25 mg/kg SC). In particular, the occurrence of episodes of licking was weakly affected by MK-801 administration, but significantly increased by CGP 43487 and APV treatment; the presence of gnawing was augmented by all the pretreatments; sniffing, locomotion, grooming and rearing occurrence were not affected by the administration of NMDA antagonists. The results suggest that the competitive antagonists which facilitated dopaminergic function without causing motor impairment could be useful supplements in the treatment of Parkinson's disease.
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  • 122
    ISSN: 1432-2072
    Keywords: Noradrenaline ; DSP4 ; Operant behaviour Timing ; Interval bisection procedure ; Acquisition ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract This experiment examined the effect of destroying central noradrenergic neurones, using the selective neurotoxin DSP4 (N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine) on the acquisition and performance of discrimination between two time intervals. Rats that had received systemic treatment with DSP4 and vehicle-treated control rats were trained in a series of discrete trials to press lever A following a 2-s presentation of a light stimulus and lever B following an 8-s presentation of the same stimulus. Both groups acquired the discrimination (〉90% correct choices) within 15 sessions; however, the DSP4-treated group showed significantly slower acquisition than the control group. When stable performance had been attained, ‘probe’ trials were introduced in which the light was presented for intermediate durations. Both groups showed sigmoid functions relating percent choice of lever B to log stimulus duration. Neither the bisection point (duration corresponding to 50% choice of lever B) nor the Weber fraction differed significantly between the DSP4-treated and control groups. The levels of noradrenaline were markedly reduced in the neocortex and hippocampus of the DSP4-treated group, but the levels of dopamine and 5-hydroxytryptamine were not altered. The results indicate that noradrenaline depletion induced by DSP4 retarded the acquisition of temporal discrimination, but did not impair steady-state discriminative precision.
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  • 123
    ISSN: 1432-2013
    Keywords: Rat ; Parotid glands ; Salivary glands ; Calcein ; Amylase ; Secretion ; Carbachol ; Noradrenalin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Effects of cholinergic and adrenergic agonists on the secretion of the fluorescent dye calcein were examined to clarify the involvement of calcium ions in the secretion of calcein from acinar cells dispersed from the rat parotid gland. Addition of carbachol (CCh) and noradrenalin (NA), but not isoproterenol (IPR), enhanced the net release of calcein from acinar cells during the subsequent 10 min in a dose range from 10−8 M to 10−6 M. The net release of calcein reached a maximum 7 min after the addition of CCh. The release of calcein was suppressed by the simultaneous additions of atropine with CCh, or phenoxybenzamine with NA. Addition of CCh induced a sustained dosedependent increase in the intracellular levels of calcium ions, ([Ca2+]i). Addition of NA at 10−6 M increased [Ca2+]i. Phenoxybenzamine completely inhibited the NA-induced increase, but propranolol did not. The removal of extracellular calcium ions did not influence the release of calcein induced by 10−6 M CCh, but it abolished the sustained increase in [Ca2+]i. The transient increase in [Ca2+]i induced by CCh was observed in the absence of extracellular calcium ions. A calcium ion chelator, 1,2-bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid (BAPTA) inhibited the CCh-induced release of calcein. The calcium ionophore, A23187 (2.5×10−6 M), but not 10−3 M dibutyryl cAMP, evoked the release of calcein. It also increased [Ca2+]i. Removal of extracellular calcium ions suppressed the A23187-induced release of calcein. These results suggest that the release of calcein from parotid acinar cells is transiently induced through an increase in [Ca2+]i by muscarinic and α-adrenergic agonists and may represent the initial process of salivary secretion.
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  • 124
    ISSN: 1432-2013
    Keywords: Diffusion coefficient ; Muscle cells ; Myoglobin ; Microinjection ; Oxygen ; Facilitated diffusion ; Intracellular oxygen transport ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract We measured the diffusion coefficient of myoglobin (D Mb) inside mammalian skeletal muscle cells with a microinjection technique. A small bolus of horse Mb was injected into a single muscle fibre and the subsequent time-dependent changes of the Mb profiles along the fibre axis were measured with a microscope-photometer. For fibres of the rat soleus muscle at 22° C, a D Mb of 1.3·10−7 cm2/s was found, confirming a result obtained previously by us for rat diaphragm muscle with a photo-oxidation technique. In the extensor digitorum longus muscle of the rat, a higher value of 1.9 · 10−7 cm2/s was measured. Auxotonic muscle contractions did not change the apparent D Mb. For the temperature range between 22 ° C and 37 ° C, a temperature coefficient, Q 10, of 1.5 was calculated. The implication of this result for the role of Mb in the facilitation of oxygen transport was examined. Model calculations show that with this relatively low D Mb value, the intracellular oxygen supply can be improved only slightly.
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  • 125
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    Pflügers Archiv 430 (1995), S. 729-738 
    ISSN: 1432-2013
    Keywords: Sleep ; Thermoregulation ; Slow wave activity ; Sleep drive ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The aim of the experiments was to study the effects of a moderate heat load on sleep in young (26-day-old) rats and to determine whether the sleep-promoting effect of heat results from stimulation of the homeostatic sleep process. The changes in sleep-wake activity, electroencephalogram slow wave activity (SWA) during non-rapid eye movement sleep (NREMS) and cortical temperature (T crt) were determined during and after long (24-h) and short (2.5-h) heat loads (elevation of ambient temperature from 26° C to 32° C), and after total sleep deprivation (SD) combined with a short-term heat load. The heat exposures elicited increases in T crt and rectal temperature (2 and 1.7° C respectively). The long-term heat load induced persistent, albeit slight enhancements in NREMS. Rapid eye movement sleep (REMS) increased with a 12-h delay during the 24-h heat load. Heat elicited an immediate large increase in SWA. After this initial increase, SWA declined and tended to fall below the baseline level during the last 12 h of the 24-h heat load. SWA and REMS were significantly suppressed after termination of 24-h heat loading. The increased SWA during the short-term heat load was not followed by subsequent alterations in sleep when the ambient temperature had returned to normal. However, after the combination of SD with the shortterm heat load the durations of NREMS and SWA were significantly enhanced compared with those found after SD at 26° C. The results are interpreted as suggesting that heat increases NREMS in the young rat by the same mechanism as is involved in the enhancement of NREMS after SD: a stimulation of sleep drive.
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  • 126
    ISSN: 1432-2013
    Keywords: Hypobaric hypoxia ; Histochemistry ; Muscle fibre-type ; Electrophoresis ; Myosin heavy chain isoform ; Soleus muscle ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Histochemical fibre-type composition and myosin heavy chain isoform component in the soleus muscle were studied in normoxic rats at postnatal ages of 5, 10, 15, and 20 weeks and in rats exposed to hypobaric hypoxia (460 torr) for 5 weeks from postnatal ages of 5, 10, and 15 weeks. The increase in the percentage of type I fibres and the concomitant decrease in that of type IIa fibres in the soleus muscle of normoxic rats were observed until 15 weeks of age. On the other hand, no change in the fibre-type composition of the muscle during postnatal development was observed in hypoxic rats, irrespective of the age at which they were exposed to hypoxia. The changes in the myosin heavy chain isoform component (MHC I and MHC IIa) of the muscle during postnatal development and by hypoxia corresponded well with those in the muscle fibre-type composition. It is concluded that hypobaric hypoxia inhibits the growth-related shift of muscle fibre-types from type IIa to type I and of myosin heavy chain isoforms from MHC IIa to MHC I in the rat soleus muscle, and that there are no changes in the muscle fibre-type composition or the myosin heavy chain isoform component caused by hypoxia after the shifts in these parameters which occur during postnatal development are completed.
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  • 127
    ISSN: 1438-8359
    Keywords: Cervical sympathectomy ; Stellate ganglion block ; Gonadotropin ; Testosterone ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract To examine the effects of bilateral cervical sympathectomy on the secretion of gonadotropin-releasing hormone (GnRH), luteinizing hormone (LH) and testosterone (TS), 24 male rats were divided into four groups: control (C), light (L), sympathectomy (S), and light-sympathectomy (LS) groups. The C and S groups were kept under a 12-h light-dark cycle and the L and LS groups were kept under continuous light for 2 weeks. After 2 weeks, blood was collected and the rats were perfused with a fixative. GnRH neurons in the hypothalamus were stained immunohistochemically, and serum LH and TS levels were measured by radioimmunoassay. Although the difference in the number of GnRH neurons between the C and S groups was not significant, the L group was significantly lower than the C or LS groups. The serum LH and TS levels in the L group were higher than in the other groups. The present results suggest that continuous light increases GnRH secretion in the hypothalamus, followed by increased secretions of LH in the pituitary and TS in the testes, and bilateral cervical sympathectomy under continuous light inhibits these hormonal changes. However, a normal circadian rhythm does not affect gonadotropin secretion. Therefore, long-term and repeated stellate ganglion block may inhibit the increases of GnRH, LH, and TS secretions induced by continuous light.
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  • 128
    ISSN: 1438-2199
    Keywords: Amino acids ; NMDA receptors ; CGS 19755 ; TCP ; Spinal cord ; Rat ; Mouse
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The possibility to visualize the NMDA recognition site with [3H]CGS 19755in vitro autoradiography was evaluated in rat brain and spinal cord sections and thereafter used to study the distribution of the NMDA recognition site in rat and mouse spinal cord. The [3H]CGS 19755 binding was concentrated to the dorsal horn, in particular to the substantia gelatinosa. Notable binding was also seen in the intermediate area and ventral horn, while some binding was observed in the funiculi. No major differences were seen in [3H]CGS 19755 binding at various levels of the rat or mouse spinal cord, although a more dense binding was seen in the mouse. A similar distribution of the [3H]CGS 19755 specific binding and the NMDA receptor associated ion-channel site, labeled with [3H]TCP, was found in the mouse spinal cord. Taken together, our data indicate that the NMDA recognition site can be visualized in rat and mouse spinal cord byin vitro [3H]CGS 19755 autoradiography.
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  • 129
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    Lasers in medical science 10 (1995), S. 273-277 
    ISSN: 1435-604X
    Keywords: Ga-As laser ; Myelinated fibre regression ; Rat ; Sciatic nerve
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Physics , Technology
    Notes: Abstract The purpose of the present study was to examine the regeneration of myelinated axons under the effect of laser irradiation at various wavelengths and energy densities. Laser irradiation at 890 nm with an energy dose of 0.33 J cm−2 as well as He-Ne laser irradiation failed to change the number of regenerating myelinated axons in distal nerve stumps on the 30th day after cutting the nerve. An increase of dose delivered to the skin to 9.33 J cm−2 resulted in a 49% decrease in the number of myelinated axons. A 24% suppression of nerve regeneration was also registered using 1220 nm wavelength laser (dose 0.98 J cm−2). This phenomenon is likely to be attributed to the stimulating effect of laser irradiation of the near-infra-red range on proliferation of fibroblasts and scar formation. 1220 nm of 7.2 J cm−2 dose effected neither the growth nor the myelinization of axons in distal nerve stumps on the 20th day following nerve damage.
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  • 130
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    Naunyn-Schmiedeberg's archives of pharmacology 352 (1995), S. 402-411 
    ISSN: 1432-1912
    Keywords: Electrogenic ion transport ; Rat ; colonic mucosa ; Somatostatin (SRIF) ; BIM-23027 ; BIM-23056 ; L-362855 ; Seglitide
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The aim of this study was to examine the potencies of several recently identified selective somatostatin (SRIF)-receptor ligands as inhibitors of electrogenic ion transport in the rat distal colonic mucosa with the view to identifying the SRIF receptor type involved. Under basal conditions, cumulative administration of SRIF and SRIF2g decreased short circuit current (SCC), a measure of electrogenic ion transport, with EC50 values of 4 nM and 9 nM respectively. The peptidase inhibitors, phosphoramidon (1 μM) and amastatin (10 μM), had no effect on the potencies of either SRIF or SRIF28. The inhibitory action of SRIF on basal SCC was suppressed by piretanide and diphenylamine-2-carboxylate, compatible with the assumption that the Na+K+2Cl− co-transporter and Cl− channels, respectively, may be involved in this antisecretory action of SRIF. Tetrodotoxin (1 μM) had no effect on the antisecretory action of SRIF, suggesting that the process was not neuronally mediated. All of the SRIF analogues examined, with the exception of BIM-23056, maximally inhibited basal SCC to a similar extent as SRIF. Seglitide and octreotide were both more potent antisecretory agents than SRIF (respective EC50 values, 0.4 nM and 1.5 nM) suggesting that this effect was mediated by a receptor belonging to the SRIF1 receptor group. The most distinguishing feature of the rank order of agonist potencies was the high potency of the selective sst2 receptor ligand, BIM-23027 (EC50, value 0.32 nM), the weaker potency exhibited by the selective sst5 receptor ligand, L-362855 (EC50 value 21 nM), and the lack of agonist activity displayed by the selective sst3 receptor ligand, BIM-23056 (EC50 value 〉 1000 nM). This profile is comparable with that observed in binding studies on the recombinant sst2 receptor. Forskolin-stimulated secretion was suppressed by SRIF analogues with the rank order of agonist potencies BIM-23027 〉 SRIF 〉 L-362855 〉 BIM-23056 which resembled that exibited under basal conditions. However, the absolute potencies of these agonists were lower (respective EC50 values 2 nM, 14 nM, 38 nM and 〉 1000 nM) whilst the magnitude of inhibition was about three fold greater. BIM-23027 and SRIF (both 30 nM) also inhibited carbachol-stimulated increases in basal SCC by 60–70%, while a similar concentration of L-362855 inhibited these responses by 11 %. BIM-23056 (1 μM) had no effect on carbachol-simulated secretion. Radioligand binding studies on rat colonic mucosal membranes using [125I]-Tyr11-SRIF suggested heterogeneity of SRIF binding sites. Thus, SRIF and SRIF28 competed for binding (IC50 values, 0.32 and 0.63 nM, respectively) with Hill slopes less than unity; while seglitide and BIM-23027 both maximally displaced only 30–40% of specific binding with apparent high affinity (respective pIC50 values, 10.1 nM and 10.0). In conclusion, SRIF decreases basal as well as both cAMP and Ca2+-dependent Cl− secretion in rat colonic mucosa. The rank order of agonist potencies suggests that receptors resembling the recombinant sst2 receptor mediate inhibition of basal and forskolin-stimulated secretion. Radioligand binding studies suggest that BIM-23027 interacts with a sub-population of [125I]Tyr11-SRIF binding sites in rat colonic mucosal membranes which probably correspond to the receptors mediating the antisecretory effects described here.
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  • 131
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    Naunyn-Schmiedeberg's archives of pharmacology 352 (1995), S. 424-428 
    ISSN: 1432-1912
    Keywords: Balloon injury ; Carotid artery ; Fibrates ; Neointima ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The inhibition of neointima formation by drugs is a major goal to prevent restenosis following angioplasty. In the present study, the effect of etofibrate on blood lipids and vessel wall was investigated using a balloon injury rat model. Two weeks after ballooning the common carotid artery neointima formation was quantified by morphometric measurement of the neointimal area and cellularity in vessel cross sections, and by fluorometric evaluation of the DNA content. Etofibrate (160 mg/kg/day) had no effect on plasma triglyceride levels, but reduced serum cholesterol by about 25%. The injury-induced increase of both the neointimal area and the DNA-content was significantly inhibited by 47% (P 〈0.005) and 34% (P 〈0.05), respectively, in the drug-treated animals in comparison to the untreated control rats. The ratio of neointima and media was significantly (P 〈 0.001) reduced from 152.9 ± 11.6% (controls) to 82.84 ± 12.59% in the etofibrate-treated group. The cellularity (numerical profile and volume density of nuclei) in the neointima was similar in both groups. In conclusion, injury-induced neointima formation is reduced in etofibrate-treated animals, which could be due to an inhibition of smooth muscle cell proliferation.
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  • 132
    ISSN: 1432-2013
    Keywords: Rat ; Hippocampus ; Hilus ; Glutamate ; Kainate ; Patch-clamp in situ
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Glial cells in the hilus of the dentate gyrus of the rat were investigated using the patch-clamp technique in acute slices of the hippocampal formation. According to their voltage-gated current patterns, two classes of glial cells — putative astrocytes and presumed glial precursor cells — were apparent. The glutamate receptor agonists kainate, glutamate, and α-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) evoked inward currents at a holding potential of −70 mV in astrocytes and presumed glial precursor cells. Inward currents could also be induced in nucleated patches, indicating a direct action on glial receptors. In presumed hilar glial precursor cells, 6,7-dinitroquinoxaline-2,3-dione (DNQX; 10 μM) blocked the kainate-induced current, while it was partially inhibited by Zn2+ (2 mM) and Evans Blue (10 μM). Cyclothiazide (100 μM), in contrast, potentiated this current, indicating the presence of AMPA receptors. In 90% of the presumed glial precursor cells the excitatory amino-acid-evoked current voltage (I/V) relations were linear or outwardly rectifying and reversed close to 0 mV, which is characteristic for non-specific cation channels. To determine the permeability to Ca2+, I/V relations were determined in a Na+-free solution containing 40 mM Ca2+ and showed reversal potentials of a wide variation ranging from −63 mV to + 1 mV with corresponding P Ca/ P Cs permeability ratios of between 0.09 and 2.10. Statistical analysis revealed that the permeability to Ca2+ significantly decreased with an advance in age (r=−0.596; n=21; P〈0.01). These data suggest that the Ca2+ influx mediated by the activation of AMPA receptors expressed in presumed hilar glial precursor cells is dependent on the developmental stage.
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  • 133
    ISSN: 1432-2013
    Keywords: Rat ; Motor unit ; Isometric contraction ; Speed
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Recordings of isometric force were obtained for twitches and (sub)maximal tetani of gastrocnemius medialis (MG) and tibialis anterior (TA) muscle units in female Wistar rats. We assessed the relationships between unit properties that have all been associated with “speed”: (1) the relative degree of peak force attained during repetitive activation at 40 Hz (P 40/P max), (2) the relative degree of final twitch fusion during the same test burst (Fus-end), and (3) various measures of the time-course of single twitches, including twitch time-to-peak and a parameter referred to as “initial fusion ratio” (Fus-in; relative decline from peak force at 25 ms from twitch onset). The various measures of twitch time-course were significantly correlated to each other with correlation coefficients varying over a fairly wide range (0.35–0.64 for MG; 0.50–0.80 for TA). Twitch time-course was also significantly correlated with Fus-end during the 40-Hz repetitive activation; the highest correlation coefficient (0.69 for MG, 0.80 for TA) was obtained for Fus-in, which was also numerically similar to Fus-end. Thus, the degree of fusion indeed seemed to be largely dependent upon aspects of twitch time-course. However, the relative degree of force mobilization obtained in the same contractions elicited by stimulation at 40 Hz was not consistently better correlated with Fus-end than with measures of single twitch time-course. Furthermore, in fast-twitch units having the same twitch time-to-peak, the force mobilization elicited by stimulation at 40 Hz (P 40 P max) was the same for MG and TA, while the degree of fusion was significantly smaller for TA than for MG units. The results demonstrate the complexity of the concept of isometric “speed” and underline the need for using several speed indicators in parallel in studies concerning the differentiation of muscle (unit) properties.
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    Pflügers Archiv 430 (1995), S. 238-245 
    ISSN: 1432-2013
    Keywords: Epilepsy ; Epileptiform activity ; Mg2+-free media ; Low magnesium ACSF ; PDS afterpotentials ; Inhibition ; Hippocampus ; Rat ; GABA ; Ca2+-dependent K+ current
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract In rat hippocampal slices epileptiform activity was induced by superfusion with Mg2+-free artificial cerebrospinal fluid (ACSF). Paroxysmal depolarization shifts (PDS) were evoked by electrical stimulation of Schaffer collaterals. To investigate the afterpotentials that follow PDS, intracellular recordings were made from CA1 pyramidal cells. The experiments revealed that several components are engaged in the generation of PDS afterpotentials in Mg2+-free ACSF. A long lasting component which determined the overall duration of the PDS afterhyperpolarization was blocked by intracellular application of ethylenebis(oxonitrilo)-tetraacetate (EGTA); concomitantly, the afterhyperpolarizations following depolarizing current injections were blocked. This indicated that the long lasting component was due to a slow Ca2+-activated K+ current. The block of Ca2+-activated K+ current uncovered a depolarizing PDS afterpotential with an N-shaped voltage dependence, suggesting that this depolarizing afterpotential component may be due to an N-methyl d-aspartate (NMDA) conductance. Intracellular injection of Cl− revealed that the PDS were followed by Cl− currents lasting about 500 ms. This component could be blocked by application of bicuculline suggesting that it is due to a synaptically GABA-mediated (i.e. γ-aminobutyric acid) Cl− current. A comparison of PDS afterpotentials in Mg2+-free ACSF and those in other models of epileptiform activity suggests that similar sequences of inhibitory components are activated in spite of different pharmacological alterations of membrane conductances which induce the epileptiform discharges.
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  • 135
    ISSN: 1573-9104
    Keywords: Hormonal status ; Legume diet ; Pisum sativum L. ; Protein turnover ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The inclusion of peas (Pisum sativum L.) as the source of protein in the diet of growing rats brings about a reduction in growth rate as well as the impairment in the liver, muscle and spleen weights as compared with casein fed controls. Also, a fall in plasma glucose, triglycerides and protein was observed in the legume fed animals, while no changes in cholesterol levels were found. Furthermore, the rats fed on the diet containing peas showed lower levels of plasma insulin, corticosterone, IGF-I and T4 as compared with casein controls. Liver and muscle total protein (mg) and total DNA (mg) were markedly decreased in the legume fed animals, but DNA/g, protein/DNA and RNA/protein ratios were similar in both dietary groups. Likewise, liver and muscle fractional synthesis rates were similar in the casein and legume groups, while the whole body protein synthesis is assumed to be lower in the legume fed animals due to differences in body weights. It is concluded that animals fed on a diet containing peas (Pisum sativum L.) as the only source of protein showed less adverse effects than those found with other legumes such asVicia faba L. orPhaseolus vulgaris L., in which protein quality, antinutritional factors and nutrient availability could be involved.
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  • 136
    ISSN: 1573-904X
    Keywords: Amphotericin B ; Amphocil® ; Fungizone® ; Colloidal Dispersion ; Tissue Distribution ; Pharmacokinetics ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract The pharmacokinetic profiles of amphotericin B (AmB) after administration of Amphocil®, an AmB/cholesteryl sulfate colloidal dispersion (ABCD) and the micellar AmB/deoxycholate (Fungizone®) were compared after repeated dosing in rats. After administration of ABCD and Fungizone at an equal AmB dose (1 mg/kg), AmB concentrations in plasma and most tissues were lower for the ABCD dose, especially in the kidneys where reduced drug concentration correlated with reduced nephrotoxicity. In contrast, AmB concentrations in the liver were substantially higher when ABCD was administered; however, without an accompanying increase in hepato-toxicity. Daily administration of ABCD for 14 days did not lead to AmB accumulation in plasma; while a slight accumulation was observed after multiple administration of Fungizone. AmB was eliminated more slowly from the plasma and various tissues and urinary and fecal recoveries of AmB were reduced after ABCD administration. These results suggest that ABCD may be stored in tissues in a form that is less toxic and is eliminated from the systemic circulation by a different mechanism than the free and protein-bound AmB in plasma. AmB accumulation in the spleen was observed when higher doses of ABCD (5 mg/kg) were administered, which could be due to saturation of hepatic uptake of AmB. Comparison of spleen concentrations of AmB between ABCD and Fungizone® at 5 mg/kg AmB doses was not possible because of Fungizone's toxicity in rats. In all other organs, AmB concentrations reached or approached a steady state within two weeks of dosing with ABCD. Urinary and fecal clearances of AmB were not different between ABCD and Fungizone administration. In summary, the distribution and elimination characteristics of AmB in rats were substantially altered when it was administered as ABCD in comparison to Fungizone. Nephrotoxicity of AmB in rats was reduced after administration of ABCD apparently because of the altered tissue distribution pattern. Thus, ABCD (Amphocil®) may be a clinically beneficial formulation of AmB in patients with systemic fungal infections.
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  • 137
    ISSN: 1573-0603
    Keywords: Endothelial cells ; Isolation ; Culture ; Mesentery ; Rat ; Rabbit
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Methods are described for the enzymatic isolation of endothelial cells from rat and rabbit mesenteric arteries and veins. The mesenteric vascular bed is incubated with an enzyme solution containing collagenase, deoxyribonuclease, papain, dithiothreitol and bovine serum albumin for 45 min at 37 °C in a shaking waterbath. After the 45 min digestion, cells are centrifuged and plated. This method yields an endothelial cell population with a high plating efficiency which is relatively free of smooth muscle contamination.
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  • 138
    ISSN: 1434-0879
    Keywords: Rat ; Pelvic ganglion ; Lactate dehydrogenase ; Isoforms ; Infravescial obstruction ; Diabetes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract We have previously shown that the intramural motor nerves in the rat bladder can function in anoxic conditions. The present study aims to explore the distribution and activity of lactate dehydrogenase (LDH), the key enzyme for ATP generation in anoxia. The activity and isoform distribution pattern of LDH was studied in pelvic ganglia from male and female rats. A histochemical investigation showed that the LDH activity was intense in the ganglion cells, and weak in the other tissue components (nerve bundles, connective tissue). The male pelvic ganglion weighed 55% more than the female pelvic ganglion, the enzyme activity per unit ganglion weight was 60% higher and the total LDH activity was 155% higher. The isoform distribution was similar, with M4 being dominant isoform, followed by M3H. Infravesical outlet obstruction in the female rat induced a threefold increase in ganglion weight, and the total LDH activity increased twofold. In this hypertrophic female ganglion a decreased relative amount of M4, and an increased amount of MH3, was found. Diabetes in the male rat had no effect on ganglion weight or its contents and isoform distribution of LDH.
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  • 139
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    European journal of applied physiology 71 (1995), S. 475-484 
    ISSN: 1439-6327
    Keywords: Cardiovascular changes ; Hyperbaric O2 ; Microspheres ; Rat ; Regional cerebral blood flow
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Hyperbaric oxygen at pressures of 300 to 500 kPa has been shown to induce changed distribution of cerebral blood flow ( $$\dot Q$$ CBF) in rats, in places reducing the supply of the supplementary O2. Thus, in the present study, the effect of hyperoxia at 101 (group 1, n = 9) and 150 (group 2, n = 9) kPa OZ on cerebral blood flow distribution and central haemodynamics was tested in conscious, habituated rats. During the control period the systolic arterial pressure (BPs), heart rate (f c), breathing frequency (f b), cardiac output ( $$\dot Q$$ c), arterial acid-base chemistry and glucose, as well as $$\dot Q$$ CBF distribution (r $$\dot Q$$ CBF) were similar in the two groups of animals. During O2 exposure, the acid-base chemistry remained unchanged. The haemoglobin decreased in group 2, but remained unchanged in group 1. The f c decreased rapidly in both groups during the change in gas composition, after which f c remained constant both in group 1 and in group 2, for whom pressure was increased. The $$\dot Q$$ c and f b decreased and BPs increased similarly in the two groups. Total $$\dot Q$$ CBF and r $$\dot Q$$ CBF decreased to the same extent in both groups, and the r $$\dot Q$$ CBF changes were equally scattered. In group 1, breathing of pure O2 did not increase the O2 supply to any cerebral region except to the thalamus and colliculi after 60 min, whereas the O2 supply to the hypothalamus decreased and remained low. In group 2, the O2 supply was unchanged compared to the control period in all regions. These findings agree with previous observations during exposures to higher O2 pressures. In air after O2 exposure the acid-base chemistry remained normal. The f c and f b increased to higher levels than during the control period. The BPs remained high. The brain blood flows were increased, inducing elevated O2 supply to several brain regions compared to the control period. In conclusion, O2 supply to the central nervous system was found to be in the main unchanged during breathing of O2 at 101 kPa and 150 kPa.
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  • 140
    ISSN: 1432-0878
    Keywords: Key words: Adrenal organ ; Atrial natriuretic peptide ; Neuropeptide Y ; Catecholamine synthesizing enzymes ; Coexistence ; Rat ; Guinea pig ; Coturnix c. japonica (Aves ; Phasianiformes) ; Lacerta viridis (Lacertilia) ; Bufo marinus ; Caldula pulchra (Anura) ; Cyprinus carpio ; Cottus scorpius (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Antisera specific for mammalian atrial natriuretic peptide (ANP) and neuropeptide Y (NPY) were applied to examine, in immunofluorescence, the occurrence of cells immunoreactive to ANP and NPY in the adrenal organs of mammals, birds, reptiles, amphibians, and bony fish. Catecholamine-containing cells were identified using antisera against tyrosine-hydroxylase, dopamine-β-hydroxylase, and phenylethanolamine-N-methyl-transferase. In all vertebrates studied, immu- noreactivities to ANP and NPY occurred in adrenal chromaffin cells but were absent from the cortex or its homolog, the interrenal. The majority of immunoreactivities to ANP and NPY was confined to the adrenaline cells. In mammals, the number of ANP-immuno-reactive cells (60%–80% of the total cell population) exceeded that of the NPY-immunoreactive cells (35%–45%). In birds, reptiles, and Amphibia, the numbers of ANP-immunoreactive (35%–40%) and NPY-immunoreactive (30%–35%) cells were in a similar range. The bony fish showed a density of both ANP-immunoreactive (80%–90%) and NPY-immunoreactive (35%–40%) cells. In all species studied, immunoreactivities to ANP and NPY partially coexisted. Generally, 30%–55% of the ANP-immunoreactive cells also contained NPY-immunoreactivity. In rat, coexistence amounted to almost 100% and in quail to 95%. Except for the rat, three subpopulations of chromaffin cells seemed to occur: ANP-immunoreactive non-NPY-immunoreactive, ANP-immunoreactive+NPY-immunoreactive, and NPY-immunoreactive non-ANP-immunoreactive cells. Thus, adrenal ANP and NPY share a conservative history and coexist as early as at the level of bony fish. The endocrine actions of ANP and NPY derived from medullary cells on cortical cells as found in mammals might be based on an ancestoral paracrine system. In submammalians, ANP and NPY may not only act as endocrine hormones, but also influence steroid-producing interrenal cells in a paracrine manner, and act as modulators on chromaffin cells.
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  • 141
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    Medical molecular morphology 28 (1995), S. 200-209 
    ISSN: 1860-1499
    Keywords: Ultrastructure ; Rat ; Endometrium ; Eosmophil ; Macrophage
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Dynamic changes in the endometrial stroma during the following 5 stages of the estrous cycle in normal rats were examined by transmission electron microscopy. Diestrus: Macrophages migrated into the endometrial stroma from blood vessels. Proestrus: Eosinophils migrated into the endometrial stroma from blood vessels. They possessed specific crystalloid granules and small granules. Estrus: The endometrial stroma was swollen and stromal cells degenerated. Eosinophils contained a few or no crystalloid granules, while the number of small granules increased. Metestrus-1: Epithelial projections protruded through the basal lamina and established focal adhesions to stromal cells. Stromal cells also adhered to one another. Metestrus-2: Most eosinophils were engulfed by macrophages. In this report, we discuss the interaction of epithelial cells with endometrial stromal cells during the normal estrous cycle.
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  • 142
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    Molecular Reproduction and Development 41 (1995), S. 331-338 
    ISSN: 1040-452X
    Keywords: Ovary ; Connexin 43 ; Gap junctions ; Cell-to-cell communication ; Follicular development ; Atresia ; Corpus luteum ; Rat ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The present immunocytochemical study examines in the rat ovary the pattern of expression of connexin 43 (Cx43), a subunit of gap junctions. Using a well-characterized specific antiserum against rat Cx43, immunoreactivity was not detected in the fetal ovary, i.e., prior to follicular formation. However, in the ovary of 20-day-old, 35-day-old, and adult rats, strong Cx43-immunore-activity was associated with the cell borders of the follicular epithelium/granulosa cells of all developmental stages (primordial follicles, preantral and antral secondary follicles). In general, immunoreactivity of the granulosa cells of large antral follicles appeared more intense than the one of smaller follicles. Staining was also seen in oocytes (cytoplasmic staining). Theca cells of large antral follicles, but not of small follicles were immunoreactive. Immunoreactive interstitial cells were not seen in ovaries of 20- and 35-day-old animals, but staining in these cells was present in adult rats. In large follicles with signs of atresia, granulosa cells lacked Cx43-immunoreactivity, whereas Cx43-immunoreactivity in their theca interna strikingly increased. Corpora lutea in the cyclic adult rats were heterogeneously stained, with either no detectable immunoreactivity, staining of cell borders of most luteal cells, or with conspicuous staining of only a few cells. In the pregnant animals on gestation days (GD) 12, 14, and 17, all luteal cells stained strongly for Cx43 at the cell surface. Shortly before delivery (GD 21), however, the staining pattern vanished and only few, presumably luteal cells remained immunoreactive. In Western blots (using homogenates of whole ovaries), the Cx43 antiserum recognized a major band of approximate Mr 43 × 103, together with minor bands, which may reflect the presence of several differently phosphorylated Cx43 forms. This is indicated by treatment with alkaline phosphatase, which reduced the banding pattern to one single band. In summary, the gap junction molecule Cx43 is abundantly expressed in all endocrine compartments of the rat ovary. The staining pattern obtained in the present study indicates that Cx43 and presumably gap-junctional communication are associated with follicular development, atresia, and the development of the interstitial gland, as well as with the development and regression of the corpus luteum. The heterogeneous staining within the ovary furthermore hints to a contribution of the local intraovarian factors in the regulation of Cx43 expression. © 1995 Wiley-Liss, Inc.
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  • 143
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    Molecular Reproduction and Development 42 (1995), S. 122-129 
    ISSN: 1040-452X
    Keywords: Cooling ; [Ca2+]i transients ; Activation ; Fertilization ; Egg ; Rat ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In mammalian eggs, activation by sperm that leads to resumption of meiosis is characterized by an explosive transient increase in intracellular calcium ion concentration ([Ca2+]i), followed by [Ca2+]i oscillations. In addition to the spermatozoon, various treatments can induce parthenogenetic activation, accompanied by an elevation of [Ca2+]i. It has been reported that cooling can induce egg activation, yet the mechanism of this phenomenon has not been elucidated. In the present study we followed changes in egg [Ca2+]i (measured by Fura-2 fluorescence ratio imaging) during activation by cooling, using conditions that ensure a low rate of spontaneous activation.Our present findings demonstrate that cooling induces egg activation as manifested by [Ca2+]i transient(s) and second polar body extrusion. Seventy-eight of 104 eggs responded to cooling with increased [Ca2+]i. Thirty-five percent of the responding eggs displayed a single [Ca2+]i transient, while 65% exhibited at least two [Ca2+]i transients within the time window of the experiment (30-40 min). Twenty-two percent of these eggs displayed high-frequency oscillations (intervals of 3.5-5.9 min). In these eggs, the overall pattern of calcium dynamics was similar to that observed in eggs activated by sperm, as judged by the transient's intervals, duration, and a gradual increase in the amplitude of successive transients. The amplitudes of [Ca2+]i transients, however, were 2-3 times lower. We propose that cooling affects [Ca2+]i homeostasis to produce fertilization-like changes in [Ca2+]i, possibly associated with parthenogenetic activation. Moreover, great care should be exercised to prevent temperature changes during egg handling. © 1995 wiley-Liss, Inc.
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  • 144
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    Developmental Dynamics 203 (1995), S. 448-455 
    ISSN: 1058-8388
    Keywords: Lens ; Immunocytochemistry ; Lens development ; Transient expression ; Glutamate decarboxylase (GAD) ; PCR ; Rat ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: We have determined the localization and developmental expression of glutamate decarboxylase (GAD67) in the rat lens. Immunofluorescence experiments showed that GAD67 was transiently expressed in the nuclear fiber cells of the lens between embryonic days (E) 15 and 20, with maximal immunostaining occurring on E17 and E18. γ-amino butyric acid (GABA) co-localized with GAD67 in the embryonic nuclear fiber cells. Reverse transcription-polymerase chain reaction (RT-PCR) tests showed that at least three alternatively spliced forms of GAD67 mRNA, including mRNAs with and without the I80 and the I86 insert, were transiently co-expressed with GAD67 in the embryonic lens. The major GAD67 protein in the lens was 67 kDa. We conclude that enzymatically active GAD67 is transiently expressed in the lens nuclear fiber cells of the embryonic rat. The transient expression is regulated by transcriptional and/or posttranscriptional processes. We speculate on the basis of possible common gene regulatory elements for glutamate and ornithine decarboxylases and the involvement of these enzymes with polyamine synthesis, that the transient expression of GAD67 may be connected to nuclear and/or DNA breakdown during lens fiber cell differentiation. ©1995 Wiley-Liss, Inc.
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  • 145
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    Microscopy Research and Technique 30 (1995), S. 319-332 
    ISSN: 1059-910X
    Keywords: Rat ; Prostate ; Epithelium ; Stroma ; Cytodifferentiation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Instructive influences of fetal mesenchyme were examined in heterotypic tissue recombinants consisting of urogenital sinus mesenchyme (UGM) from male and female rats and distal ductal tips from adult rat prostate. Tissues were grown under the renal capsule of male hosts for periods up to 28 days. Resultant growths exhibited typical prostate histology. Expression of lobe-specific proteins for the ventral (prostatic steroid binding protein [PSBP]) lateral (seminal vesicle secretion II [SVS II]), and dorsal prostate (secretory transglutaminase [TGase]) were examined by immunocytochemistry. Male or female UGM combined with terminal segments of the ventral or dorsal prostate and immunolabeled with antibodies to lobe-specific proteins demonstrated expression of all three secretory products. The pattern of staining was consistent with a compound inductive response from the UGM. Unique to this study was our ability to use a defined mesenchymal tissue (female ventral mesenchymal pad [VMP]). This tissue is specifically associated with ductal branching morphogenesis and cytodifferentiation of the ventral prostate. Distal ductal tips from the dorsal lobe of the adult male prostate when recombined with female VMP and grown in vivo exhibited transformation of secretory phenotype, and the epithelium expressed mRNAs for PSBP. Immunocytochemistry of serial sections did not demonstrate labeling for TGase in the new epithelial growth. Ultrastructural analysis of the heterotypic recombinants indicated that the epithelium had similar characteristics to those of normal ventral prostate. Early stages of the mesenchymal-epithelial interactions resulted in dedifferentiation of the adult epithelium to solid cords of stratified cells. These findings illustrate the potent instructive capacity of a defined fetal UGM to influence development and cytodifferentiation of adult prostate epithelium. © 1995 Wiley-Liss, Inc.
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  • 146
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    Microscopy Research and Technique 30 (1995), S. 366-380 
    ISSN: 1059-910X
    Keywords: Myogenesis ; Myosin ; MyoD ; Myogenin ; PDGF ; FGF ; Transferrin ; Chicken ; Rat ; C2 cells ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The myogenic precursor cells of postnatal and adult skeletal muscle are situated underneath the basement membrane of the myofibers. It is because of their unique positions that these precursor cells are often referred to as satellite cells. Such defined satellite cells can first be detected following the formation of a distinct basement membrane around the fiber, which takes place in late stages of embryogenesis. Like myoblasts found during development, satellite cells can proliferate, differentiate, and fuse into myofibers. However, in the normal, uninjured adult muscle, satellite cells are mitotically quiescent. In recent years several important questions concerning the biology of satellite cells have been asked. One aspect has been the relationship between satellite cells and myoblasts found in the developing muscle: are these myogenic populations identiacal or different? Another aspect has been the physiological cues that control the quiescent, proliferative, and differentiative states of these myogenic precursors: what are the growth regulators and how do they function? These issues are discussed, referring to previous work by others and further emphasizing our own studies on avian and rodent satellite cells. Collectively, the studies presented indicate that satellite cells represent a distinct myogenic population that becomes dominant in late stages of embryogenesis. Moreover, although satellite cells are already destined to be myogenic precursors, they do not express any of the four known myogenic regulatory genes unless their activation is induced in the animal or in culture. Furthermore, multiple growth factors are important regulators of satellite cell proliferation and differentiation. Our work on the role of one of these growth factors [platelet-derived growth factor (PDGF)] during proliferation of adult myoblasts is further discussed with greater detail and the possibility that PDGF is involved in the transition from fetal to adult myoblasts in late embryogenesis is brought forward. © 1995 Wiley-Liss, Inc.
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    Comparative clinical pathology 5 (1995), S. 177-182 
    ISSN: 1433-2981
    Keywords: T3 ; T4 ; Progesterone ; Rabbit ; Rat ; ELISA ; Automation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract We have designed and executed comparative studies with the objective of selecting instrumentation for the non-isotope immunological determination of free and total thyroxine (fT4 and tT4), and total triiodothyronine (tT3) and progesterone in rat plasma for general toxicity studies. In addition, this instrumentation has been used for the determination of progesterone in rabbits for early pregnancy diagnosis in reprotoxicity studies. During instrument selection, special emphasis has been given to automation and on-line coupling capabilities, maximal flexibility in method development, walkaway capability, manufacturers' computer validation and GLP performance, linearity and intra-assay CV. For tT3 and tT4, seven instruments have been compared with each other whereas for progesterone and fT4, three instruments were compared. Normal rat plasma values have been subjected to variance analysis, followed by Duncan testing. The instrument selection process finally indicated the ES300 Enzymun ELISA of Boehringer Mannheim as the best candidate on the aspects defined above. Spiking recovery values on the ES300 are presented. The ES300 provides a reliable pregnancy diagnosis for rabbits as early as day +4 after mating in the predosing period.
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    Comparative clinical pathology 5 (1995), S. 237-250 
    ISSN: 1433-2981
    Keywords: Clinical chemistry ; Haematology ; Neonate ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract This study was performed to collate background data for a range of blood pathology parameters in neonatal rats, strain Crl: CD BR VAF/Plus, which could be used to assess organ maturity and function. This information was considered necessary as concern over neonatal toxicity has been expressed by scientists in the pharmaceutical, agrochemical and industrial fields. Haematological and clinical chemistry profiles were generated from neonate blood samples, taken via cardiac puncture. Samples were obtained, under terminal anaesthesia, on days 4, 12, 15 and 20 post partum. Analyses were performed on a regime of pooled and individual samples per sex for each litter. All results were compared with normal blood parameter ranges for non-pregnant rats aged approximately 9–10 weeks. The haematological profile indicated that the pups had an immature haemopoietic system and were developing subclinical but physiological anaemia in the early postnatal period. This was shown by low and decreasing Hb concentration and MCHC, a large proportion of reticulocytes in the red cell mass and low RBC, PCV, total and differential WBC. APTT was considerably shorter in the neonate, whereas PT was longer. Fibrinogen concentration was low. Principal findings from the clinical chemistry profile indicated apparent immaturity of the liver, kidneys and adrenal cortex. In the time course observed GPT, albumin, globulins, sodium and chloride increased; potassium, urea and bilirubin decreased; AP, calcium, phosphates, triglycerides and cholesterol levels were high compared with normal adult ranges. Both profiles showed there to be no obvious differences between the male and female pups up to 20 days post partum.
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    Comparative clinical pathology 5 (1995), S. 13-24 
    ISSN: 1433-2981
    Keywords: Clinical chemistry ; Haematology ; Lactation ; Pregnancy ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Clinical pathology parameters are a valuable index relating to the pathophysiological state of an animal and are routinely measured in most toxicological studies. In order to interpret blood data in reproductive studies it is first necessary to know ‘normal’ background ranges through pregnancy and lactation. The purpose of this study was to generate this database using the Crl:CD VAF/Plus strain of rat as a model. Blood profiles were generated by bleeding time-mated female rats at various intervals during the pre- and postnatal period (days 7,12,15 and 20 of pregnancy, days 4,12,15 and 20 lactation). A routine set of clinical pathology analyses were performed. The haematology results showed that during pregnancy an increase in plasma volume causes a reduction in haemoglobin concentration, RBC and PCV leading to the onset of ‘emergency haematopoiesis’ and hence an increased reticulocyte count. There was also a decline in circulating WBC, mainly lymphocytes. Both the APTT and PT increased during gestation. With the exception of WBC, the haematology values returned to within normal non-pregnant ranges during lactation. The clinical chemistry results indicated that organ function was changed during gestation and lactation in the dam compared to that of a normal non-pregnant female. These changes were primarily linked to hypertrophy of the liver, changes in hydration and an altered renal threshold.
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    Comparative clinical pathology 5 (1995), S. 98-101 
    ISSN: 1433-2981
    Keywords: Dehydrogenases ; Electron microscope ; Immunisation ; Rat ; Thymus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The distribution and localisation of lactate dehydrogenase (LDH) and succinate dehydrogenase (SDH) were studied using electron microscopy (EM) and cytochemical reactions in the thymus of rats 3 days afterEscherichia coli immunisation. In thymic lymphoblasts of untreated rat thymus, LDH was present mainly in the nuclear envelope, in the rough endoplasmic reticulum (RER), sometimes in the cytoplasm and in the mitochondria, whereas the SDH reaction product was evident in the nuclear envelope and in the mitochondria. In the lymphocytes the LDH and SDH reaction product was observed in some mitochondria and in small quantities in the nuclear envelope. A small amount of LDH reaction product was also present loosely distributed within the cytoplasm. In the treated rats the LDH and SDH localisation was similar to that of the untreated rats, but the amount of both reaction products was increased in the nuclear envelope. Since the lymphocytes both in treated and untreated animals showed small amounts of LDH and SDH reaction product compared to that observed in the lymphoblasts, our results show a correlation between the level of cell maturation and the distribution of LDH and SDH reaction product in the thymocytes of both treated and untreated rats. However, the increase of LDH and SDH in the nuclear envelope of thymocytes of treated animals indicates a variation of cell metabolism afterE. coli immunisation. This finding would suggest that the nuclear envelope is a probable site of enzyme synthesis.
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    Comparative clinical pathology 5 (1995), S. 189-195 
    ISSN: 1433-2981
    Keywords: Cell lines ; Guinea pig ; Human ; Hypolipaemic agents ; Peroxisome proliferators ; Rat ; Species difference
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Peroxisomes are ubiquitous organelles of eukaryotic cells and are present in significant amounts in hepatic liver cells. Peroxisomal enzymes contribute to several metabolic pathways including fatty acid, purine and amino acid catabolism or bile acid synthesis. The peroxisomal oxidative reactions produce hydrogen peroxide, mostly degraded by catalase which prevents oxidative stress. Moreover, peroxisomes are involved in arylderivative drug detoxification through its epoxide hydrolase activity. In rodents the exposure of cells to xenobiotic compounds such as fibrates, phthalates/adipates and chlorophenoxyacetic acid derivatives, which are used as hypolipaemic drugs, plasticizers and pesticides respectively, lead to a liver mass increase and to a high peroxisome proliferation. This latter event is due to a strong genetic activation triggered by the PPAR (peroxisome proliferator activated nuclear receptor). Human contrasts with rodent since there is no, or little, effect of the above cited compounds. In contrast, the defect of single or multiple peroxisomal functions caused by genetic disorders lead to an increase of very long chain fatty acid level, which is toxic, especially for brain and kidney. The liver response to xenobiotics of the peroxisome proliferator class may be modulated by auxiliary compounds such as hormones (e.g. thyroid hormone) or nutriments (e.g. retinoids).
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  • 152
    ISSN: 1433-2965
    Keywords: Absorptiometry ; Dual-energy X-ray absorptiometry ; Histomorphometry ; Ovariectomy ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Dual-energy X-ray absorptiometry (DXA), together with the use of ultra-high resolution software, recently appeared as an accurate method for determining bone mineral density (BMD) in the rat. In order to assess the ability of this technique to detect changes in bone mass in the rat rapidly and precisely, we measured BMD at various sites of the femur using DXA subregional analysis. In particular, we studied the BMD of the metaphyseal part of the femur (M-BMD) rich in trabecular bone, and compared the values obtained with the cancellous bone volume measured by histomorphometry. In short-term ovariectomized animals (experiment 1), M-BMD was the only parameter to differentiate statistically between 10 ovariectomized (OVX) and 10 SHAM-operated (SHAM) rats (−11.2%,p〈0.01) 9 days after surgery. M-BMD still expressed the greatest variation between OVX and SHAM rats 42 days following ovariectomy (experiment 2) (−16.1%,p〈0.001 v −6.2%,p〈0.01 for the total femur BMD) and confirmed previous data demonstrating a greater loss of cancellous than cortical bone after cessation of ovarian activity. M-BMD was highly correlated with cancellous bone volume (BV) in normal (r=0.82,p〈0.001,n=30), OVX (r=0.77,p〈0.001,n=22) and SHAM (r=0.88,p〈0.001,n=21) rats. Furthermore, subcutaneous treatment with rat parathyroid hormone fragment (1–34) (r-PTH(1-34)) partially and significantly protected animals from trabecular osteopenia induced by OVX; there was a similar degree of protection of BV and M-BMD (50% and 61% respectively), while BMD of the entire femur achieved complete protection. This M-BMD measurement, specifically reflecting cancellous bone mass as confirmed by the correlation study and the response to PTH treatment, is a sensitve and simple method which can be used to assess any precocious modifications of bone density under physiopathological or therapeutic conditions in experimental rat models of bone loss.
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  • 153
    ISSN: 1433-2981
    Keywords: APTT ; Dog ; Human ; Monkey ; Mouse ; PT ; Rabbit ; Rat ; Stability ; Storage
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Changes in plasma activated partial thromboplastin times (APTT) and prothrombin times (PT) in mice, rats, rabbits, dogs, monkeys and human were examined for up to 96 h at storage temperatures of 4 and 25°C. Prolongation of APTT in rats was rapid and marked, with times doubling within 24 h post-sampling. Plasma APTT of human and monkey were also affected, but to a lesser extent. No effect was observed in mice, rabbits and dogs. On the other hand, the magnitude of PT changes was much smaller than that observed with APTT in all species. No significant differences were noted between the results from samples stored at 4°C or 25°C for either test. The false prolongation of APTT is clearly undesirable in a toxicity study, especially in rats. It is important therefore to minimise these changes by performing this test under strict time-controlled conditions.
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  • 154
    ISSN: 1432-0878
    Keywords: Adrenal organ ; Atrial natriuretic peptide ; Neuropeptide Y ; Catecholamine synthesizing enzymes ; Coexistence ; Rat ; Guinea pig ; Coturnix c. japonica (Aves, Phasianiformes) ; Lacerta viridis (Lacertilia) ; Bufo marinus, Caldula pulchra (Anura) ; Cyprinus carpio, Cottus scorpius (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Antisera specific for mammalian atrial natriuretic peptied (ANP) and neuropeptide Y (NPY) were applied to examine, in immunofluorescence, the occurrence of cells immunoreactive to ANP and NPY in the adrenal organs of mammals, birds, reptiles, amphibians, and bony fish. Catecholamine-containing cells were identified using antisera against tyrosine-hydroxylase, dopamine-β-hydroxylase, and phenylethanolamine-N-methyl-transferase. In all vertebrates studied, immunoreactivities to ANP and NPY occurred in adrenal chromaffin cells but were absent from the cortex or its homolog, the interrenal. The majority of immunoreactivities to ANP and NPY was confined to the adrenaline cells. In mammals, the number of ANP-immuno-reactive cells (60%–80% of the total cell population) exceeded that of the NPY-immunoreactive cells (35%–45%). In birds, reptiles, and Amphibia, the numbers of ANP-immunoreactive (35%–40%) and NPY-immunoreactive (30%–35%) cells were in a similar range. The bony fish showed a density of both ANP-immunoreactive (80%–90%) and NPY-immunoreactive (35%–40%) cells. In all species studied, immunoreactivities to ANP and NPY partially coexisted. Generally, 30%–55% of the ANP-immunoreactive cells also contained NPY-immunoreactivity. In rat, coexistence amounted to almost 100% and in quail to 95%. Except for the rat, three subpopulations of chromaffin cells seemed to occur: ANP-immunoreactive non-NPY-immunoreactive, ANP-immunoreactive+NPY-immunoreactive and NPY-immunoreactive non-ANP-immunoreactive cells. Thus, adrenal ANP and NPY share a conservative history and coexist as early as at the level of bony fish. The endocrine actions of ANP and NPY derived from medullary cells on cortical cells as found in mammals might be based on an ancestoral paracrine system. In submammalians, ANP and NPY may not only act as endocrine hormones, but also influence steroid-producing interrenal cells in a paracrine manner, and act as modulators on chromaffin cells.
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  • 155
    ISSN: 1432-0878
    Keywords: Enkephalin ; Opioid peptides ; Spleen ; Innervation ; Neuro-immunology ; Species differences ; Immunohistochemistry ; Cow ; Pig ; Guinea-pig ; Mouse ; Rat ; Dsungarian hamster
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The opioidergic innervation of the mammalian spleen and possible species differences were investigated. Light-microscopic immunohistochemistry revealed that splenic nerves of bovine and porcine spleen, but not of rat, mouse, hamster and guinea-pig spleen contained proenkephalin-derived opioidergic innervation. Immunoreactivity to both prodynorphin and pro-opiomelanocortin was absent from splenic nerves. In bovine and porcine spleen, fibers immunoreactive for met-enkephalin, met-enkephalin-Arg-Phe, met-enkephalin-Arg-Gly-Leu, leu-enkephalin and peptide F formed perivascular plexus, traveled in trabecular connective tissue, and extended into the capsule. Spatial relationships with immune cells were apparent in the white and red pulp, excluding lymphoid follicles. Colocalization of enkephalin immunoreactivity with immunoreactivities for tyrosin hydroxylase, dopamin-β-hydroxylase, and neuropeptide Y, but not for substance P or calcitonin gene-related peptide were found. Our results provide evidence that opioid expression in splenic innervation is strongly species-dependent and exclusively proenkephalin-derived. Colocalization with marker enzymes of noradrenergic neurons indicates a mainly postganglionic sympathetic origin of proenkephalinergic splenic innervation. Opioidergic perivascular nerves probably control the splenic blood flow. A close interrelationship of opioidergic fibers with immune cells provides the anatomical basis for direct effects of neurally released opioids on splenic immune functions.
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  • 156
    ISSN: 0003-276X
    Keywords: Sertoli cell ; Testis ; Morphometry ; PTU ; Rat ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Background: The testes of rats treated neonatally with propylthiouracil (PTU) grow to almost twice their normal size. The cause of testicular enlargement has been suggested to be the result of delayed maturation of Sertoli cells, allowing Sertoli cell division to occur beyond the 15th postnatal day, the commonly recognized cutoff date for Sertoli cell divisions. It has been shown that an increased population of Sertoli cells in postnatal development supports increased numbers of germ cells in adult animals. After examining developing rats treated neonatally with PTU, we hypothesized that an approximate 10-day delay in maturation was occurring and proceeded to test this hypothesis experimentally. Thus the purpose of this report was to determine if a 10-day delay in maturation could explain the increased numbers of Sertoli cells and increased testis size in PTU-treated animals.Methods: Both control animals and animals treated neonatally with PTU N = 5/group were sacrificed at 15 and 25 days of age and prepared for electron microscopy.Results: Micrographs show and morphometric ultrastructural analysis of numerous parameters demonstrated at the 95% probability level that Sertoli cells from 25-day-old PTU animals are not different in size and most constituents (volume and surface area) from 15-day-old control animals and are less mature than 25-day-old control animals. Mitosis of Sertoli cells was observed in PTU-treated animals in 25-day-old animals but not in agematched controls. The number of Sertoli cells in 25-day-old PTU-treated animals is significantly increased over age-matched controls. Micrographs show the presence of immature Sertoli cell nuclei in 25-day-old animals receiving PTU as well as increased germ cell degeneration in this group. Sertoli cell tight junction formation is also delayed in PTU-treated animals as compared with controls.Conclusions: Together, the data show that delayed maturation of Sertoli cells occurs in treated animals that corresponds to a minimum of 10 developmental days. In the immature state, Sertoli cells continue to divide. Data presented herein and published data related to PTU treatment indicate that delayed maturation of the Sertoli cell results in delayed maturation and proliferation of other testicular cell types. From this and from published data, the hypothesis is presented that the Sertoli cell is responsible for the overall control of testis development. © 1995 Wiley-Liss, Inc.
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  • 157
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    The @Anatomical Record 241 (1995), S. 113-122 
    ISSN: 0003-276X
    Keywords: Lymph node ; Reticular fibers ; Collagen fibrils ; Reticular cells ; Rat ; Scanning electron microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Background: The reticular framework in the lymph node has in the past been studied mainly by light microscopy of silver-impregnated specimens. The aim of the present study is to understand three-dimensionally the ultrastructure and organization of the reticular framework better than before.Methods: The mesenteric lymph nodes of the rat were prepared either an alkali-water maceration method or a conventional method and were observed in a scanning electron microscope (SEM).Results: The SEM study of alkali-water macerated tissues visualized directly the reticular fiber network in the lymph node. The reticular fibers consisted of thin bundles of collagem fibrils. They were continuous with the collagen fibriliar sheaths of blood vessels and lymphatic sinuses as well as with the fibrous capusule, thus acting as a skeleton of the lymph node. The arrangement of the reticulum was variable, depending on individual compartments. The SEM study of conventionally treated tissues, on the other hand, clarified the shape of reticular cells and their relationship with the reticular fibers. The sinus reticular cells connected with the sinus lining cells but separated from the parenchymal reticular cells, indicating that the former two originate from lymphatic endothelial cells. The parenchymal reticular cells varied in shape depending on their locations but essentially shared features with fibroblasts.Conclusions: The arrangements of the reticular fibers in the parenchyma were closely related to the associated reticular cells, showing the possibility that the reticular cells maintain the shape of the reticular framework suitable for each compartment of the lymph node. © 1995 Wiley-Liss, Inc.
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  • 158
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    The @Anatomical Record 241 (1995), S. 529-540 
    ISSN: 0003-276X
    Keywords: Cell division ; Differentiation ; Growth and development ; Parotid gland ; Rat ; Salivary gland ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Background: In contrast to the considerable amount of research that has been done on the proliferative activity of the several types of parenchymal cells in the developing submandibular glands of rodents, systematic studies of cellular proliferation in the developing parotid gland have been confined to the acinar cells. The purpose of the present study was to attempt to fill this knowledge gap.Methods: Tritiated thymidine was parenterally administered to Sprague-Dawley rats at ages representative of the pre- and postnatal development of the parotid gland, and glands were harvested for autoradiography 90 min after injection. Mitotic activity among all cell types was verified by electron microscopy.Results: At all ages, the % labeled cells was much greater among the acini than any other cell type, including well-differentiated cells at 25 and 40 days. However, there were only small alterations in the proportions of cells comprised by the major cell types.Conclusions: Current theories on the histogenesis of salivary glands and their neoplasms are based on the renewing population model, in which both normal differentiated cells and neoplastic cells arise from undifferentiated stem cells in the ducts. However, these results suggest that most of the migration and redifferentiation in the developing rat parotid gland must be in the opposite direction, i.e., the acinar cells redifferentiate into ductal cells. They also indicate that until there are precise data on the rates of cell death among the several cell types, it remains more appropriate for salivary glands to be categorized as an expanding, rather than renewing, population. © 1995 Wiley-Liss, Inc.
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  • 159
    ISSN: 0003-276X
    Keywords: Pseudorabies virus ; Retrograde tracing ; Sacral parasympathetic nucleus ; Nitric oxide synthase ; Choline acetyltransferase ; Immunohistochemistry ; Rat ; Female ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Background: The purpose of this study was to elucidate parasympathetic preganglionic neurons in the spinal cord that project axons in pathways to the uterus and to reveal their neurotransmitter phenotype.Methods: “Uterine-related” neurons were identified by using a combination of retrograde axonal tracers: (1) Fluorogold injected into the ganglia of termination of preganglionic fibers, and (2) a transganglionic axonal tracer (pseudorabies virus) injected into the uterus. Immunohistochemistry was used to reveal virus-labeled neurons and their neurotransmitter marker.Results: Double-labeled (Fluorogold + pseudorabies virus) “uterine” preganglionic neurons were identified in the sacral parasympathetic nucleus of the rat lumbosacral spinal cord. Subpopulations of neurons in the sacral parasympathetic nucleus were shown to be immunoreactive for choline acetyltransferase or nitric oxide synthase. Double-staining immunohistochemistry (for pseudorabies virus + neurotransmitter enzyme) revealed that some of the uterine-related preganglionic neurons were cholinergic and some nitric oxide synthase-containing.Conclusions: These results demonstrate a subpopulation of pregauglionic parasympathetic neurons in the sacral parasympathetic nucleus that are involved in uterine innervation. In addition, both acetylcholine and nitric oxide could be used to modify activity in the postganglionic neurons, which directly innervate the uterus. © 1995 Wiley-Liss, Inc.
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  • 160
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    The @Anatomical Record 241 (1995), S. 579-584 
    ISSN: 0003-276X
    Keywords: Somatostatin mRNA ; Spinal trigeminal nucleus ; Dorsal horn ; Spinal cord ; In situ hybridization histochemistry ; Rat ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Background: The spinal trigeminal nucleus and the dorsal horn of the spinal cord are the critical areas in relaying noxious impulses. They also contain large amounts of somatostatin-like immunoreactivities. Early publications focussed on immunohistochemical studies and were devoid of a detailed description of the distribution of somatostatin in the two nuclei at the molecular level.Method: Frontal tissue sections from the mudulla and the spinal cord of eight rats were examined and a non-radioactive in situ hybridization histochemical procedure was adopted to study the distribution of somatostatin mRNA positive neurons in the two nuclei.Results: A widespread distribution of somatostatin mRNA containing neurons was shown in the two nuclei at all levels. The positive neuron profiles were normally round or oval in shape and small to medium in size. Three types of cells were identified, which were associated with the intensity of the hybridization signals. The highest density of somatostatin mRNA positive neuron profiles was found in the gelatinous subnucleus at the caudal part of the spinal trigeminal nucleus and in the substantia gelatinosa of the dorsal horn at the levels of the cervical and lumbar cords. Most of them belonged to type I neurons.Conclusions: The present findings are compared to reports of previous studies. It is suggested that somatostatin in the two nuclei may play an important role in the modulation and transmission of pain signals. © 1995 Wiley-Liss, Inc.
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  • 161
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    The @Anatomical Record 242 (1995), S. 1-10 
    ISSN: 0003-276X
    Keywords: Annexin ; Cell Death ; Calcium ; Phosphatidylserine ; Transglutaminase ; EGF ; Inflammation ; Rat ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Background: Enigmatically, degradation of debris generated in programmed cell death (apoptosis) elicits little inflammation. Having previously detected the upregulation of lipocortin 1 (LC1), a 35-kDa protein with anti-inflammatory and immuno-suppressive properties, at sites of non-inflammatory phagocytosis in the central nervous system (J Neurosci Res 36:491-500, 1993), we sought to determine if LC1 was involved in apoptosis.Methods: LC1 immunoreactivity in mammary glands of adult rats was quantified in situ using video microdensitometry before and during postlactational regression.Results: LC1 is present in the mammary ducts but is absent from the alveoli during lactation. One day after weaning, however, LC1 is detected in the lactiferous cells and, as apoptosis proceeds over the ensuing 4 days, total LC1 in the gland increases 〉10-fold over resting levels. LC1 remains high in both the apoptotic cells and epithelial phagocytes through day 10, but the total LC1 per gland drops as the apoptotic cells are cleared.Conclusions: Published experiments have shown that LC1 specifically binds Ca++ and phosphatidylserine, and that these affinities are modulated by tyrosine phosphorylation and cross-linking with transglutaminase. Thus, LC1 appears to be a candidate for several putative activities in apoptosis (e.g., phagocyte recognition via phosphatidylserine binding and/or buffering intracellular Ca++) in addition to its anti-inflammatory role. © 1995 Wiley-Liss, Inc.
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  • 162
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    The @Anatomical Record 242 (1995), S. 188-194 
    ISSN: 0003-276X
    Keywords: Skeletal Muscle ; Cardiac Muscle ; Regeneration ; Transplantation ; Rat ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Background: The ability of skeletal muscle to regenerate after injury is well established. In contrast, cardiac muscle is incapable of regeneration and recovery after injury. The aim of the present study was to evaluate and compare the regeneration pattern of cardiac and skeletal muscle after transplantation into a skeletal muscle bed in rats.Methods: The following group of transplants were performed at the site prepared by removing the host extensor digitorum longus (EDL) muscle. The first group consisted of cardiac muscle transplanted as one piece or after mincing into 1-mm pieces. The second group included cotransplants of cardiac and skeletal muscle minces that were intermixed. Entire EDL muscle or minced EDL muscle were also transplanted for comparison. Rats were sacrificed 3-30 days after transplantation for morphological analysis.Results: The results demonstrated that skeletal muscle transplants underwent rapid regeneration, and by 30 days the entire muscle was filled with regenerated myofibers. In transplants of cardiac muscle significant inflammation, myocardial degeneration and necrosis were observed. In spite of the necrosis and fibrosis, the presence of a few regenerated myotubes in the outer region was observed. In cardiac and skeletal muscle cotransplants, the inflammation was restricted to cardiac tissue; however, by 30 days the entire contransplant was filled with regenerated myotubes and myofibers.Conclusions: These results show that skeletal muscle is capable of growth, regeneration, and integration with the cardiac muscle after cotransplantation. Combination of skeletal and cardiac muscle may prove useful in defining the cellular processes necessary for enhancing cardiac repair after injury. © 1995 Wiley-Liss, Inc.
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  • 163
    ISSN: 0003-276X
    Keywords: Osteogenesis in vitro ; Sex-steroids ; Glucocorticoid ; Differentiation ; Rat ; Chicken ; Bone ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Glucocorticoids and sex-steroids can modulate osteogenesis in vivo and in vitro. Although the effects of glucocorticoids on bone cells in vitro have been described in detail, the role of sex-steroids is not as well defined. We examined whether sex-steroids influence bone metabolism indirectly by regulating glucocorticoid effects on bone. Interactions of the sex-steroid progesterone or its analog RU38486 with the glucocorticoid dexamethasone (dex) were studied in functional assays of osteogenesis. Three osteoblastic models were evaluated:(1) the rat bone marrow stromal cell (RBMC) nodule system; (2) the chick periosteal osteogenesis (CPO) model; and (3) ROS 17/2.8 cells. RU38486, progesterone, and unlabelled dex competitively inhibited 3H-dex uptake by ROS 17/2.8 cells as well as its (3H-dex) binding to cytosol preps.Both RU38486 and progesterone inhibited dex-induced increases in alkaline phosphatase in CPO cultures, in RBMC cultures, and in ROS 17/2.8 cells. Dex-induced decreases in cell proliferation in ROS 17/2.8 cells were reversed by RU38486 but dex-induced increases in proliferation in the CPO model were not affected. In CPO cultures, dex-induced increases in collagen synthesis were inhibited completely by RU38486 and progesterone, Dex-dependent nodule formation in the RBMC was blocked by RU38486. Both RU38486 and dex mediated reduction of calcium uptake in the CPO model but did not affect mineralized tissue area.The data indicate that RU38486 and progesterone competitively inhibit dex-mediated stimulation of osteogenesis in vitro; this inhibition is exerted on early but not late stage differentiation events of osteoprogenitor cells. © 1995 Wiley-Liss, Inc.
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  • 164
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    The @Anatomical Record 242 (1995), S. 531-544 
    ISSN: 0003-276X
    Keywords: Pulmonary veins ; Pulmonary circulation ; Corrosion casting ; Scanning electron microscopy ; Pulmonary edema ; Lymphatics ; Rat ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Background: Pulmonary lymphatics are critical to clearing lung fluid. Although their structure can be shown with light and transmission electron microscopy, scanning electron microscopy of their casts can better show their number, size, shape, distribution, and degree of filling. This technique has identified four forms of lung lymphatics, but these forms have not been fully evaluated by tissue microscopy. A most important site of pulmonary edema formation, the pulmonary capillary, is just upstream from small veins which have focal, smooth muscle tufts termed venous sphincters. Because of their constricting potential, these sphincters may control lung perfusion and cause edema.Methods: With light and transmission electron microscopy of tissue and scanning electron microscopy of casts, the lymphatic forms were explored in relation to the tissue anatomy in rats without pulmonary edema and with mild-to-moderate edema caused by extended vascular rinsing.Results: The edematous lungs had increased sacculo-tubular lymphatics adjacent to the venous sphincters. These lymphatics were in the adventitial connective tissue and were partially endothelialized. As lymphatics became more tubular their endothelium became more complete. Collagen fibers traversed the lumen of these lymphatics even where endothelial cells were present and caused the lines on the surface of the lymphatic casts. Overlapping endothelial cells caused clefts on the casts.Conclusions: Scanning electron microscopy of lymphatic casts better defines their ultrastructure and shows the spatial relationship of veins and their sphincters to venous lymphatics. Sphincter contraction may influence pulmonary lymph production which could affect other aspects of regional lung perfusion. © 1995 Wiley-Liss, Inc.
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  • 165
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    The @Anatomical Record 242 (1995), S. 562-565 
    ISSN: 0003-276X
    Keywords: Rat ; Brain ; Microcirculation ; Development ; Pericyte ; Electron microscopy ; Gap junction ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Background: Fine structural study revealed the intercellular coupling between the pericyte and the endothelial cells via the gap junctions, in the capillaries of the basal forebrain of rat embryos.Results: Gap junctions were constructed by the adluminal plasmalemma of pericyte and the abluminal plasmalemma of endothelial cells.Conclusions: Gap junctions are membranous channels that directly join the cytoplasms of the pericyte and endothelial cell and imply some substantial role for the pericyte on the endothelial proliferation. It is postulated that the function of the pericyte in the prenatal mammals are assigned to the regulation of the development of cerebral microcirculation. © 1995 Wiley-Liss, Inc.
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  • 166
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    The @Anatomical Record 243 (1995), S. 175-185 
    ISSN: 0003-276X
    Keywords: Rat ; Bone histomorphometry ; Femoral neck ; Aging ; Ovariectomy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Background: This study characterizes the changes in cortical and cancellous bone and cross sectional moment of inertia of the femoral neck from aging and ovariectomized (ovx'd) rats to determine their role in the previously reported ovx-induced reduction of mechanical strength in the femoral neck.Methods: Undecalcified double-fluorescent lableled cross sections of femoral neck of 3.5-, 5.5-, 6.5-, and 8.5-month-old female rats and rats ovx'd at 3.5 months for 2, 3, and 5 months of 45 rats were studied. The estimated endocortical and trabecular surfaces, cortical and cancellous bone histomorphometry, and cortical moment of inertia were determined.Results: The femoral neck was adding cortical bone between 3.5 and 5.5 months of age by increasing cortical thickness and decreasing marrow cavity area. No change of cortical bone mass was found between 5.5 and 8.5 months and the same amount of cancellous bone was observed between 3.5 and 8.5 months of age. Ovariectomy-induced cancellous, but not cortical bone loss. The loss was due to a transient ovx-induced negative bone balance that by 5 months post-ovx produced a 42% loss in trabecular bone while the histomorphometry profiles were the same as controls. The crosssectional moment of inertia increased with age but did not differ significantly between ovx'd and controls.Conclusions: Our findings suggest that the ovx-induced cancellous bone loss could be a contributing factor to the reduced mechanical strength in the femoral neck of ovx'd rats reported previously. © 1995 Wiley-Liss, Inc.
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  • 167
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    The @Anatomical Record 243 (1995), S. 223-233 
    ISSN: 0003-276X
    Keywords: Rat ; Lymph node ; Innervation ; Age ; EM ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Background: Earlier studies have shown that the innervation of axillary lymph nodes is increased in old animals, compared to juvenile rats. The question thus arose, whether changes are also observable at an ultrastructural level for varicosities which form the actual transmission units.Methods: Ensheathed (axons) and unensheathed axonal profiles (open areas), were quantified with their spatial relationships to cells at the ultrastructural level in the axillary lymph nodes of juvenile ( 〈 6 weeks) and old ( 〉 2 years) Wistar rats.Results: In both groups of animals the majority of open areas, irrespective of their content of vesicles, lie at distances of more than 1,000 nm from cells in the plane of the section. Almost all open areas, located within 1,000 nm of a cell, are related to reticular cells, a small minority to plasma cells, and even fewer to lymphocytes. In the old animals there is a highly significant increase in the total number of sections of open areas with and without vesicles per unit area of medulla. The number of open areas related to cells does not change significantly in the old animals, as also the percentage of profiles containing vesicles. Only the percentages of open areas with and without vesicles related to plasma cells increase significantly in the old animals. This change is paralleled by a highly significant increase in the percentage volume of plasma cells in the old animals. On average, the profiles related to cells are closer to the cells in the older animals.Conclusions: The observed changes indicate that the innervation of lymph nodes of the rat is absolutely increased in the old animals, but that the relations between the various categories of profiles and their relations to cells tend to remain constant. © 1995 Wiley-Liss, Inc.
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  • 168
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    The @Anatomical Record 243 (1995), S. 261-271 
    ISSN: 0003-276X
    Keywords: N-CAM ; HNK-1 ; Neural crest cell ; Rat ; Heart ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Background: Neural cell adhesion molecule (N-CAM) is important in the migration of neural crest cells and is expressed in the developing heart. The pattern of expression of N-CAM in the heart of early rat embryos was investigated to shed light on the potential role of N-CAM in cardiac neural crest cell migration.Methods: N-CAM expression was studied by immunohistochemistry in Sprague-Dawley rat hearts between embryonic days 11.5 and 15.5 HNK-1 immunoreactivity was also investigated for comparison with that of N-CAM.Results: A continuity of N-CAM immunoreactivity was transiently detected from the outflow tract to the recurrent nerve. N-CAM was also expressed around the sinus venosus, inferior vena cava, sinotrial septum, and coronary sinus, as well as on mesenchymal cells in the atrioventricular endocardial cushion tissues.Conclusions: The continuous N-CAM immunoreactivity from the outflow tract to the recurrent nerve appeared to represent the pathway along which cardiac neural crest cells migrate. N-CAM-immunoreactive sites around the sinus venosus may correspond to migrating neural crest cells that differentiate into nerve fibers or cardiac ganglia. Results indicate that N-CAM may play an important role in the migration, proliferation, and transformation of neural crest cells, thereby contributing to cardiac morphogenesis and to innervation around the heart and great arteries. © 1995 Wiley-Liss, Inc.
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  • 169
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    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 243 (1995), S. 519-523 
    ISSN: 0003-276X
    Keywords: Hippocampal formation ; Mossy fibers ; Electron microscopy ; Granule cells ; Dense-core Vesicles ; Neuropeptide ; Rat ; Cholecystokinin ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Background: The possibility that mossy fiber endings in the rat hippocampal formation may contain cholecystokinin (CCK) was reexamined.Methods: For this, CCK-immunoreactivity was examined by light and electron microscopy using the avidin-biotin complex method.Results: At the light level, the topographical distribution of perikarya and processes with CCK-like immunoreactivity (CCK-LI) was similar to that previously described by others. Ultrastructural analysis of the dentate gyrus and CA3 region of the hippocampus revealed that some mossy fiber terminals contained CCK-LI most often affiliated with large, dense-core vesicles (DCV). Quantitative analysis revealed that 4-8% of the mossy terminal profiles examined (n = 350) contained CCK-labeled DCVs, which corresponded to 0.03-0.2 labeled DCVs per 100 μm2 of neuropil.Conclusions: The presence of CCK-LI within mossy fibers in the rat suggests that there is less species variability in peptide expression in this pathway than formerly believed. © 1995 Wiley-Liss, Inc.
    Additional Material: 3 Ill.
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  • 170
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    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 241 (1995), S. 181-204 
    ISSN: 0003-276X
    Keywords: Testis ; Morphometry ; Germ cells ; Spermatogenesis ; Rat ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Background: There has never been a study of the components of germ cells as they progress through spermatogenesis.Method:The structural changes taking place in rat germ cells, from spermatogonia to late supermatids, were studied utilizing morphometric techniues conducted largely at the ultrastructural level.Results:Volume and surface area parameters for virtually all cellular and subcellular features were obtained for nine periods during the spermatogenic cycle. Virtually of germ cell components show dynamic properties associated with specific phases of their development.Conclusions: The data provided can be used in an objective wey to characterize structural changes to funcational properties of germ cells. © 1995 Wiley-Liss, Inc.
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  • 171
    ISSN: 0003-276X
    Keywords: Actin filament ; Myoid cell ; Sertoli cell ; Testis ; Development ; Cryptorchid ; Rat ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Background: Abundant actin filaments are present in myoid cells and Sertoli cells in the testis. In the adult rat, the filaments form a lattice arrangement within the myoid cell, and show a hexagonal pattern in the basal junctional regions of Sertoli cells.Methods: Isolated seminiferous tubules and frozen sections were prepared from juvenile to adult Wistar rat testes, stained with FITC-conjugated phalloidin, and observed by confocal microscopy. Unilateral cryptorchidism was induced in adult rats, and seven days later, their testes were also examined.Results: In the myoid cell, parallel actin filaments running circularly around the seminiferous tubules were observed at 15 and 20 days of age. Then, at 30 days, actin filaments arranged longitudinally along the tubular long axis appeared in addition to the circular bundles. A lattice arrangement of actin-filament bundles in myoid cells became obvious at 40 days, when elongated spermatids are found in the tubule. Actin filaments in the basal junctional regions of Sertoli cells did not acquire the hexagonal pattern seen in the adult testis until 30 days of age. In the cryptorchid testes, the arrangement of actin filaments in the both cells showed a remarkable change compared to the control testis; the filaments became thinner and disrupted.Conclusions: A lattice arrangement of the actin filaments in the myoid cell appear at around 30 days, before the completion of spermatogenesis. A hexagonal pattern of the filaments in the junctional regions of Sertoli cells has already developed at this age. Cryptorchidism affects the actin filaments of the both cells. © 1995 Wiley-Liss, Inc.
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  • 172
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    The @Anatomical Record 241 (1995), S. 377-382 
    ISSN: 0003-276X
    Keywords: Magnetic resonance ; imaging ; Magnetic resonance ; spectroscopy ; Gastric mucosa ; Mucus ; Acid secretion ; Prostaglandins ; Pentagastrin ; Rat ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Background: Morpho-functional studies of gastric mucosa are hampered by the lack of a technique allowing direct in situ visualization of the mucus in small living laboratory animals.Methods: The material covering the gastric surface was studied in vivo in rats by magnetic resonance imaging (MRI) at 4.7 Tesla, and modification of its secretion was evaluated after pharmacological treatment.Results: In unstimulated animals, the glandular portion of the stomach was lined by a layer of material emitting a signal of high intensity. Administration of 16,16-dimethyl prostaglandin E2 caused an accumulation of this material within a maximum 30 min after the administration of the drug. At 45 min, gastric emptying occurred and at 60 min, the lumen was almost free of material emitting a signal of high intensity. An increase in the intensity of the signal emitted from the material filling the gastric lumen was found after pentagestrin injection. After 45 min, the intensity of the signal emitted from the material in the gastric lumen decreased. 1H localized spectroscopy showed that after injection of pentagastrin there was an increase in the water proton peak within the gastric lumen. About one hour after stimulation, the water proton peak returned to the basal value.Conclusions: This study demonstrates that MRI displays gastric mucus in living small rodents and represents a sensitive screening test for pharmacological action on this structure, enabling morpho-functional studies. © 1995 Wiley-Liss, Inc.
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  • 173
    ISSN: 0003-276X
    Keywords: Rat ; Bone ; Osteoblast ; Osteocyte ; Gap junctions ; Vimentin ; Immunolocalization ; Ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Background: The immunogold labeling technique and transmission electron microscopy were used to demonstrate the expression and position of the intermediate filament vimentin in rat osteoblast and osteocyte cell bodies and cell processes. Conventional light and transmission electron microscopic studies of bone cells demonstrated adjacent cell linkage to be mediated by osteoblast and osteocyte processes present within the canalicular system traversing the bone matrix. The cell processes were filled with densely packed filaments, many of which have been shown previously to be actin microfilaments. The appearance, however, of 10 nm diameter filaments in some cell processes and the fact that the intermediate filament vimentin has been defined in many cells of mesenchymal origin raised the possibility that some of these filaments might be vimentin. The ultrastructural colloidal gold immunochemical technique allowed for demonstration in situ of the expression of vimentin filaments plus accurate definition of their position.Methods: The studies were performed in newborn rat femoral and tibial diaphyseal cortical bone and in 1-week-old repair bone from 2.4 mm diameter defects made through the lateral cortex in 6-week-old rat femurs and tibias. The bone tissues for the immunochemical study were fixed in 1% glutaraldehyde, 4% paraformaldehyde, and 0.1 M phosphate buffer (pH 7.4) for 2 days. Decalcification was performed in 6% EDTA for 2-3 days. Infiltration involved use of Lowicryl resin K4M, and the embedding and curing processes were performed in a cryostat with temperatures -30°C. An antivimentin monoclonal antibody was used for labeling using the postembedding technique. Effective antibody dilutions ranged from 1:10 to 1:200, with the dilutions of 1:25 and 1:100 showing the best combination of filament labeling with the least matrix background. The grids were exposed to 10 nanometer gold colloid conjugated goat anti-mouse IgM for demonstration of binding.Results: Vimentin immunolabeling was defined clearly in relation to filaments within the osteoblast and osteocyte cell body cytoplasm, throughout the entire length of the osteoblast and osteocyte cell processes, and in close relationship to the intercellular gap junctions which were present within the cell processes both close to the cell bodies and within the canaliculi well away from them.Conclusions: Immunogold labeling demonstrates the presence of the intermediate filament vimentin in osteoblast and osteocyte cell bodies and processes of rat bone. Vimentin distribution is not concentrated to specific areas, is present throughout the extent of the bodies and processes, and is seen immediately adjacent to gap junctions. © 1995 Wiley-Liss, Inc.
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  • 174
    ISSN: 0003-276X
    Keywords: Fatty acid-binding proteins ; Immunocytochemistry ; Ovary ; Postnatal development ; Gonadotropins ; Rat ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Background: The ovary of adult rats expresses two types of cytoplasmic fatty acid binding proteins (FABP), i.e., Heart FABP (H-FABP) and intestinal 15 kDa proteins (I-15p). We studied immunohistochemically the cellular localizations of these FABPs in the ovaries of rts at various postnatal ages and in the ovaries of immature (3-week-old)rats treated with pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin (HCG).Methods: The cryosections of ovaries were incubated with polyclonal antibodies aganist H-FABP and I-15P, and the immunoreactions were visualized at both light and electron microscpic levels.Results: The immunorectivity for H-FABP occurred temporarilly in tthe follicular epithelian (granulosa) cells from 3 days to 2 weeks post partum, and then was localized exclusively to the theca/interstitial gland cells from 2 weeks to adulthood. In contrast, the immunoreactivity for I-15P appeared temporarily in a small subsct of theca/intersitial gland cells from 2 to 3 weeks, disappeared at 4 weeks, and was localized exclusively to the corpus luteum cells after the onset of ovulation in the animal around 5 weeks. In the immature rat ovaries induced to ovulate by treatment with gonadotropins, I-15P-immunocreative cells were first recognized in the luteinized granulosa layer of large preovulatory follicles, and increased in number progressively in the developing corpora lutea after the ovulation.Conclusions: Two type of FABPs are expressed in ditinct steroid-producing cell types of rat ovary, and their expressions seem to be regulated in results suggest that FABPs play specifie roles in the ovarian hormone synthesis. © 1995 Wiley-Liss, Inc.
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  • 175
    ISSN: 0003-276X
    Keywords: Cattle ; Freemartin ; Human ; Rat ; Gubernaculum ; Processus vaginalis ; Testis descent ; Sexual differentiation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Background: Freemartinism occurs in some speices of ruminants and affects most female bovine fetuses in hterrosexual, multiple pregnancies owing of susion of the chorionic blood circulations soon after implantation. Maldevelopment of the ovaries and Müllerian ducts have been described and recognized as resulting from exposure of their respective primoridia to an excess of anti-Müllerian hormone. The Present study aimed to analyse the prenatal growth the development of the gubernaculum in freemartins to find out its pssible affliction through foetal testis hormones derived from their male co-twin.Methods: Histolgical sections of young and frawings and photographs of further developed freemartins and conrol male and female bovine foeuses were analysed. The specimens had been collected ealier for analsis of the time course of male and female gonadal and gential development and its impairment associated with freemartinism.Results: The gubernaculum of 35-40 day-old male and female fetuses was in the intial stage of development and of similar appearance in all specimens. Gubenacula of 60-70-day-old male fetuss differed from those of females of similar age in various respects: the male gubernaculum size was larger and extension of the processus vaginalis was deeper. Freemartins showd and intermediate development with some individuals resembling male and othes resembling female agemates. During furher development, gubernacula in males developed into muscular cremaster sacs, whereas those in females generally did not develop beyond the size and structural complexity of 70-day-old foetuses. Beyond day 70 of fetal life, gubernaculum development in freemartins definitly showed male characteristics with respect to size and growth of a processus vaginalis with a cremaster muscular wall. The male-like pattern of the outgrowth of the processus vainalis changed during the second half of prenatal life. Rather than its further deepening as in mals, this structure became inveted to become emerging as a papilla-like structure from the inguinal abdomen bottom. An explanation is proposed for this unprecedented inversion, taking into account: (1) the faster and higher reaching rightsided ascent of the kidneys and gonads, (2) the femalelike outgrowth of the cranial gonadal suspensory ligaments, and (3) the absence of scrotum development. The ovaries and mesonephric remnants in developing freemartins, during their ascent together with the kidneys while remaining attached to the bottom of the developing processus vaginalis sacs via the gubernaculum ligament, are proposed to act together to pull up the bottom of the processus vaginalis sacs. From this action, “inverted hernia sacs” result as the irreversible consequence.Conclusion: The data support the concept that foetal testes act, via as an yet unidentified third hormone, to establish malelike development of gubernacula into muscular cremaster sacs. Further work is required to reveal the identity of this hormone. Furthermore, the apparent similarity of the freemartins' inverted processus vaginalis sacs and the fetal rodents' gubernacular cones suggests that the ruminants' and rodents' processus vaginalis are essentially similar structures. Thus there is no longer an urgent need to distinguish between two different types of gubernaculum development and testis descent in rodents and ruminants, respectively, and involving or not fetal gubernacular cones. The present observations may thus contribute to the development of a unified hypothesis for sexually dimorphic development of the gubernaculum throughout the mammalian class. © 1995 Wiley-Liss, Inc.
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  • 176
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    The @Anatomical Record 241 (1995), S. 255-267 
    ISSN: 0003-276X
    Keywords: Rat ; Diabetic pregnancy ; Embryonic dysmorphogenesis ; Neuroepithelium ; Blood cells ; Mitochondria ; High-amplitude mitochondrial swelling ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Background: Previous studies in vivo and in vitro have suggeted that the oxidative metabolism of the embryo may have a role in the treatogenicity of diabetic pregnancy. In particular, the production of reactive oxygen species by the embroyonic mitochondiria has been implicated in the teratological process. The induction of congenital malformations by the diabetic milieu occurs during the early embryonic development. The present study aimed to estimate the role of the embryonic mitochondria in the teratological process of diabetic pregnancy by studying mitochondrial morphology in the embryos exposed to a diabetic environment in vivo or in vitro during early organogenesis and late fetal development.Methods: For studies in vivo embryos of control or streptozotocin-dia-betic rats were taken at gestational days 9-11 and sujected to light and electron microscopical analysis. The brain, heart, and liver of day-15 fetuses were also observed. For studies in vitro day-9 embryos of normal rats were cultured in a whole-embryo culture system for 48 hours. The culture media were supplied with high conectration of diabetes-related substrates and metabolites, and their effect on structure of embryonic neuropeithelial cells determined.Results: The light microscopoical observations demonstrated numberous cytoplasmic vaculoes in the ectoderm of day-9 embryos and the neuroepithelium and blood cells of day-10 and day-11 embryos of diabetic rats. Ultrastructuraily, these vacuoles were found to be mitochondria undergoing large-amplitude swelling with loss of matrix density and disturbed cristae. In contrast, no Mitochondrial differences were found in the brain, heart, and liver, when day-15 fetuses from normal and diabeic rats were compared. Ultrastructural analysis of day-9 embryos cultured for 48 hours in the presnce of high conectrations of D-Glucose, pyruvate, β-hydroxybutyrate, and α-ketoisocaproate also showed high-amplitude mitochondrial swelling in the neuroepithelium. The motochondrial swelling was, however, not found in embryos cultured in a high conectration of L-glucose, excluding simple osmotic effects of the diabetes-related substrated and metabolites.Conclusions: The mitochondrial morphological changes appeared in embryos subjected to a diabetic environment during a time period when the congenital malformations in diabetic pregnancy are induced. The results support the notion that embryonic mitochondria are involved in the teratological process of diabetic pregnancy. © 1995 Wiley-Liss, inc.
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  • 177
    ISSN: 1432-0878
    Keywords: Key words: Enkephalin ; Opioid peptides ; Spleen ; Innervation ; Neuro-immunology ; Species differences ; Immunohistochemistry ; Cow ; Pig ; Guinea-pig ; Mouse ; Rat ; Dsungarian hamster
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The opioidergic innervation of the mammalian spleen and possible species differences were investigated. Light-microscopic immunohistochemistry revealed that splenic nerves of bovine and porcine spleen, but not of rat, mouse, hamster and guinea-pig spleen contained proenkephalin-derived opioidergic innervation. Immunoreactivity to both prodynorphin and pro-opiomelanocortin was absent from splenic nerves. In bovine and porcine spleen, fibers immunoreactive for met-enkephalin, met-enkephalin-Arg-Phe, met-enkephalin-Arg-Gly-Leu, leu-enkephalin and peptide F formed perivascular plexus, traveled in trabecular connective tissue, and extended into the capsule. Spatial relationships with immune cells were apparent in the white and red pulp, excluding lymphoid follicles. Colocalization of enkephalin immunoreactivity with immunoreactivities for tyrosin hydroxylase, dopamin-β-hydroxylase, and neuropeptide Y, but not for substance P or calcitonin gene-related peptide were found. Our results provide evidence that opioid expression in splenic innervation is strongly species-dependent and exclusively proenkephalin-derived. Colocalization with marker enzymes of noradrenergic neurons indicates a mainly postganglionic sympathetic origin of proenkephalinergic splenic innervation. Opioidergic perivascular nerves probably control the splenic blood flow. A close interrelationship of opioidergic fibers with immune cells provides the anatomical basis for direct effects of neurally released opioids on splenic immune functions.
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