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  • Articles: DFG German National Licenses  (177)
  • 1995-1999  (177)
  • 1995  (177)
  • Rat  (177)
  • Nuclear reactions
  • Numerical Methods and Modeling
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  • Articles: DFG German National Licenses  (177)
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  • 1995-1999  (177)
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  • 101
    ISSN: 1432-1106
    Keywords: Infrared image ; Somatosensory cortex ; Skull ; Rat ; Gerbil
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Infrared images of the skull surface were obtained in urethane-anesthetized rats and gerbils before, during and after mechanical stimulation of the face and mystacial vibrissae on one side. Areas of increased temperature on the skull, localized mainly over the face area of the primary somatosensory cortex contralateral to the side of stimulation, appeared within 4–5 s after the onset of stimulation. Rarely, such temperature change was recorded bilaterally. Temperatures did not remain high on the intact skull in rats, but fell to baseline within minutes after stimulus onset regardless of stimulus duration. In rats in which the skull had been thinned and in gerbils with intact skull, temperatures remained elevated during the course of stimulation. We were unable to resolve the activation of individual vibrissae.
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  • 102
    ISSN: 1432-1106
    Keywords: Pain ; Nociception ; Sensorimotor integration ; Receptive field ; Somatosensory ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The withdrawal reflex pathways to hindlimb muscles have an elaborate spatial organization in the rat. In short, the distribution of sensitivity within the cutaneous receptive field of a single muscle has a spatial pattern that is a mirror image of the spatial pattern of the withdrawal of the skin surface ensuing on contraction in the respective muscle. In the present study, a search for neurones encoding the specific spatial input-output relationship of withdrawal reflexes to single muscles was made in the lumbosacral spinal cord in halothane/nitrous oxide-anaesthetized rats. The cutaneous receptive fields of 147 dorsal horn neurones in the L4-5 segments receiving a nociceptive input and a convergent input from A and C fibres from the hindpaw were studied. The spatial pattern of the response amplitude within the receptive fields of 118 neurones was quantitatively compared with those of withdrawal reflexes to single muscles. Response patterns exhibiting a high similarity to those of withdrawal reflexes to single muscles were found in 27 neurones located in the deep dorsal horn. Twenty-six of these belonged to class 2 (responding to tactile and nociceptive input) and one belonged to class 3 (responding only to nociceptive input). None of the neurones tested (n=20) with reflex-like response patterns could be antidromically driven from the upper cervical cord, suggesting that they were spinal interneurones. With some overlap, putative interneurones of the withdrawal reflexes to the plantar flexors of the digits, the plantar flexors of the ankle, the pronators, the dorsiflexors of the ankle, and a flexor of the knee, were found in succession in a mediolateral direction. It is concluded that neurones that are able to encode the specific spatial input-output organization of the withdrawal reflexes to single muscles do exist in the deep dorsal horn. Such reflex encoders appear to have a “musculotopic” organization. A hypothesis of the organization of the withdrawal reflex system is presented.
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  • 103
    Electronic Resource
    Electronic Resource
    Springer
    Experimental brain research 106 (1995), S. 79-92 
    ISSN: 1432-1106
    Keywords: Autonomic ; Axon collaterals ; Medial tegmental field ; Motoneurons ; Respiratory ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Stimulation of the caudal raphe nuclei alters visceral functions. The caudal raphe nuclei project to the nucleus of the solitary tract, which receives the central terminations of vagal afferents and plays an important role in the central integration of autonomic activities. The caudal raphe nuclei also project to the somatic and preganglionic autonomixc motoneurons of the spinal cord. Diamidino yellow was injected into the nucleus of the solitary tract, and fast blue was injected into either the cervical, thoracic, or lumbar spinal cord. Large numbers of double-labeled neurons were present within the caudal raphe nuclei and the adjacent reticular formation of the medial tegmental field. This observation documents that individual raphespinal and reticulospinal neurons project an axon collateral to the nucleus of the solitary tract. These data demonstrate the anatomic substrate for global modulation of the autonomic motoneuron pool by the caudal raphe nuclei.
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  • 104
    ISSN: 1432-1106
    Keywords: Neural transplantation ; Locomotion ; Paw shaking ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The present study was designed to examine the effects of an intraspinal transplantation of embryonic brainstem neurons on fictive motor patterns which can develop in hindlimb nerves of adult chronic spinal rats. Seventeen adult rats were spinalized at T8-9 level and, 8 days later, a suspension of embryonic cells obtained either from the raphe region (RR, n=8) or from the locus coeruleus (LC, n=9) was injected caudally (T12–13) to the cord transection. Eight control animals (control rats) were spinalized and injected with vehicle under the same conditions. One to three months later, the animals were decorticated and fictive motor patterns were recorded in representative hindlimb nerves. The data revealed that both control and grafted spinal rats could exhibit two distinctly different fictive motor patterns, one which could be associated with stepping and the other with hindlimb paw shaking. They further showed that following transplantation of embryonic RR or LC neurons the excitability of the spinal stepping generator was increased, whereas that of the spinal neural circuits which generate hindlimb paw shaking was not significantly affected. A histological analysis performed on the spinal cord segments below the transection revealed complete absence of serotonin and noradrenaline immunoreactivity in control spinal animals and, in both types of grafted rats, an extensive monoaminergic reinnervation with synaptic contacts between monoaminergic transplanted neurons and host interneurons and/or motoneurons. The possible mechanisms by which grafted monoaminergic neurons can influence the spinal motor networks are discussed.
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  • 105
    ISSN: 1432-1106
    Keywords: Spreading depression ; GFAP ; Astrocytes ; Focal ischemia ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract This study investigated astroglial responses after focal cerebral ischemia in the rat cortex induced by photothrombosis. Astrocyte activation was studied at various time points by immunocytochemistry for glial fibrillary acidic protein (GFAP) and vimentin (VIM). We found a dual astrocytic response to focal ischemia: In the border zone of the infarct, GFAP-positive astrocytes were present within 2 days and persisted for 10 weeks. These astrocytes additionally expressed VIM. Remote from the ischemic lesion, cortical astrocytes of the entire ipsilateral hemisphere transiently expressed GFAP, but not VIM, beginning on day 3 after photothrombosis. This response had disappeared on day 14. By recording DC potentials, five to seven spreading depressions (SD) could be detected on the cortical surface during the first 2 h after photothrombosis. Treatment with MK801, a non-competitive NMDA-receptor antagonist, completely abolished SD and remote ipsilateral astrocytic activation, while the reaction in the border zone of the infarct remained unchanged. Functionally, persistent astrocytosis around the infarct might be induced by leukocyte-derived cytokines, while NMDA-receptor-mediated SD might cause remote responses.
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  • 106
    ISSN: 1432-1106
    Keywords: Respiratory rhythm ; Respiratory neurons ; Pacemaker neurons ; Perforated patch ; Nystatin ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract In brainstem-spinal cord preparations isolated from newborn rats, intrinsic burst-generating properties of preinspiratory (Pre-I) neurons in the rostral ventrolateral medulla, which have been suggested to be primary respiratory rhythm-generating neurons, were studied by “perforated” whole-cell recordings using the antibiotic nystatin. Nystatin causes small pores to be formed in the cells, through which pass small monovalent ions. For blockade of chemical synaptic transmission, perfusate Ca2+ concentration was lowered to 0.2 mM and the Mg2+ concentration was increased to 5 mM. In Iow-Ca2+, high-Mg2+ solution (referred to here as “low Ca”), 10 of 55 Pre-I neurons generated rhythmic bursts (burst type), 14 fired tonically (tonic type), and 31 were silent (silent type). Burst-type neurons showed periodic depolarization of 5–12 mV in low Ca, at a rate of 12±6.5/min. Hyperpolarization of the membrane caused decrease in or disappearance of the periodic depolarization and prolongation of the cycle period. Thus, the burst generations were voltage dependent. The firing frequency of tonictype neurons was 2.3±1.6 Hz and was decreased by hyperpolarization. In 6 of these neurons, the firing patterns changed to burst patterns during continuous hyperpolarization. Membrane depolarization by continuous outward current injection into some silent-type neurons (3 of 11 tested) induced bursting activity. Activity of C4 and Pre-I neurons was completely silent with 0.1–1 μM tetrodotoxin (TTX) added to the standard perfusate. In low Ca, burst-type neurons (n=3) were also silent with 1 μM TTX perfusion. Inspiratory neurons either became silent (n=4) or fired tonically (n=1) in low Ca. The present study by “perforated” whole-cell recordings confirmed that some Pre-I neurons possess intrinsic burst-generating properties, which were not attributable to phasic synaptic inputs.
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  • 107
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    Experimental brain research 106 (1995), S. 106-110 
    ISSN: 1432-1106
    Keywords: Spiking neural network ; Refractory period ; Phase transition ; Finite-size effect ; Hippocampal culture ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Electrophysiological properties of neurons as the basic cellular elements of the central nervous system and their synaptic connections are well characterized down to a molecular level. However, the behavior of complex noisy networks formed by these constituents usually cannot simply be derived from the knowledge of its microscopic parameters. As a consequence, cooperative phenomena based on the interaction of neurons were postulated. This is a report on a study of global network spike activity as a function of synaptic interaction. We performed experiments in dissociated cultured hippocampal neurons and, for comparison, simulations of a mathematical model closely related to electrophysiology. Numeric analyses revealed that at a critical level of synaptic connectivity the firing behavior undergoes a phase transition. This cooperative effect depends crucially on the interaction of numerous cells and cannot be attributed to the spike threshold of individual neurons. In the experiment a drastic increase in the firing level was observed upon increase of synaptic efficacy by lowering of the extracellular magnesium concentration, which is compatible with our theoretical predictions. This “on-off” phenomenon demonstrates that even in small neuronal ensembles collective behavior can emerge which is not explained by the characteristics of single neurons.
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  • 108
    ISSN: 1432-1106
    Keywords: Retina ; NMDA ; HRP ; Neurotoxicity ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract To establish a new behavioral animal model of excitotoxicity, we injected adult rats intraocularly with a single dose of 2, 20, or 100 nmol of N-methyl-d-aspartate (NMDA). We quantified visual impairment by plotting the size of the visual field in which the rats successfully oriented towards a small, moving target. In comparison to the saline-injected (contralateral) control side, the side injected with 2 nmol of NMDA was not significantly impaired. When injected with higher doses, the rats were nearly blind immediately after surgery, with only about 20% (20 nmol NMDA) or 10% (100 nmol NMDA) of residual vision. Within about 3 weeks, however, visual performance returned to near-normal levels. Simultaneous intraocular administration of a non-competitive NMDA-antagonist, MK-801 (1 nmol), resulted in complete behavioral protection. NMDA administration led to a dose-dependent loss of cells within the ganglion cell layer, as assessed in whole-mounted retinae which were retrogradely labelled with horseradish peroxidase (HRP). Whereas 2 nmol of NMDA led to the loss of about 30% of retinal ganglion cells (RGCs), at higher NMDA doses only 13% of the RGCs survived. After the injection of 20 nmol of NMDA, large-diameter RGCs (〉22 μm) survived the lesion to a greater extent than small diameter cells (8–21 μm); at 100 nmol cells of all diameters were equally affected. The number of Nissl-stained cells with small diameters (〈11 μm), presumed to be displaced amacrine cells, was also affected by NMDA, although to a lesser degree. Analysis of behavioral performance (vision score) and the number of cells in the retina revealed a correlation of r=0.76 between visual performance and the number of HRP-filled RGCs immediately after surgery. Lower correlations were found between visual performance and cells stained with Nissl of diameters smaller than 11 μm (presumably displaced amacrine cells) or larger than 11 μm (presumed RGCs without retinofugal connections; r=0.55 and r=0.58, respectively). Because of the spontaneous recovery of vision, all correlations declined to values near 0 after 3 weeks. Thus, despite a dramatic loss of RGCs following NMDA administration, visual deficits recover significantly in adult rats within 2–3 weeks.
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  • 109
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    Experimental brain research 106 (1995), S. 145-155 
    ISSN: 1432-1106
    Keywords: Adaptation ; Depression ; LTD ; LTP ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Synaptic depression was assessed from intracellular recordings in cortical tissue slices. Evoked postsynaptic potentials exhibited synaptic depression with an exponential or double exponential decrease (time constants: 〈1–30 s) in amplitude during repetitive afferent stimulation by short trains of suprathreshold stimuli. Depressed synaptic responses recovered with an exponential time course (time constants: 10 s-8 min) during presentation of similar short trains of stimuli every 5 or 10 s. Cortical cells recorded extracellularly in cat visual cortex show similar time constants of response decrement during adaptation to moving stripes. Postsynaptic voltage- or ion-regulated conductances and chloride conductances do not appear to be involved in synaptic depression. Input resistance changes and effects of injection of chloride indicate a lack of GABAA receptor-mediated effects. Hyperpolarizing or depolarizing neurons, and pairing polarization with afferent stimulation, also did not affect synaptic depression. This distinguishes these processes from long-term depression and long-term potentiation. Our results suggest that the most likely mechanisms of synaptic depression and adaptation in cortical cells are presynaptic decrease in transmitter release and/or receptor desensitization. Short-term postsynaptic changes may also occur after synaptic depression.
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  • 110
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    Springer
    Experimental brain research 106 (1995), S. 391-402 
    ISSN: 1432-1106
    Keywords: Taste ; Insular cortex ; Excitatory amino acid receptor ; Iontophoresis ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Two main subclasses of ionotropic receptors for excitatory amino acids (EAAs), N-methyl-d-aspartate (NMDA) receptors and non-NMDA receptors, are involved in neurotransmission in the cortex of mammals. To examine whether EAAs are transmitters at the cortical taste area (CTA) in rats and to elucidate which types of the two ionotropic receptors operate at these synapses, we studied the effects of microiontophoretic administration of EAA antagonists on the responses of 64 taste cortical neurons to four basic taste stimuli in urethane-anesthetized rats. Both d-2-amino-5-phosphonovalerate (APV), a selective antagonist for NMDA receptors, and 6-cyano-7-nitro-quinoxaline-2,3-dione (CNQX), a selective antagonist for non-NMDA receptors, suppressed most of the taste responses. The percentage of neurons suppressed by APV (70.3%) was almost the same as that suppressed by CNQX (64.1%). These suppressive effects were independent of the effects of background discharges during the prestimulus, water-rinsing period. The percentage of neurons suppressed by the antagonists did not differ between any pairs of taste stimuli. The number of neurons possessing both receptors was larger in the granular insular area (area GI), one of the two CTAs, than in the dysgranular insular area (area DI). In addition, taste responses were suppressed by CNQX or by both APV and CNQX in area GI in a significantly larger number of layer V neurons than in area DI. The present results indicate that normal excitatory transmission of taste afferents in the CTA in rats was mediated by both NMDA and non-NMDA receptors. The finding that a large fraction of neurons in the CTA in rats mediated taste information through NMDA receptors in normal transmission might be related to the higher potency of the plasticity observed in the CTA.
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  • 111
    ISSN: 1432-0738
    Keywords: Key words 5-Lipoxygenase inhibitors ; N-Hydroxyureas ; Nephrotoxicity ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  The N-hydroxyurea derivatives 70C ((E)-N-{3-[3-(4-fluorophenoxy)phenyl]-1-(R,S)-methylprop-2-enyl}-N-hydroxyurea) and its (R) 225C and (S) 404C enantiomers, which were being developed as 5-lipoxygenase inhibitors for the treatment of certain allergic and inflammatory conditions, were found to cause severe glomerulonephropathy in the rat. The lesion appeared to be of greater severity in female rats compared with male rats. In addition, 70C and 225C treated animals appeared more severely affected than 404C treated animals. Detailed examination of the lesion in animals dosed with 225C showed that there was a clear relationship between the onset of the lesion and the dose given, i.e. the higher the dose the sooner the lesion developed. The earliest changes detected in the kidney by transmission electron microscopy were noted in the glomeruli, in which the visceral cells appeared enlarged and showed varying degrees of foot process loss. In the more advanced lesion, the degree of foot process loss became more obvious and changes in the kidney tubules were seen by light microscopy. The morphological changes were mirrored by a dose-related increase in water consumption, an increased kidney to body weight ratio and gastrointestinal oedema, suggesting impaired renal function. Shortly after the onset of foot process loss, decreases in the total plasma protein and albumin and increases in the plasma cholesterol, triglycerides, urea and creatinine were recorded. These changes, particularly the foot-process loss, together with increased proteinuria, hypoalbuminaemia, hypercholesterolaemia and lipaemia, are all characteristic of “minimal change nephrotic syndrome”. Because of the serious nature of the kidney lesion caused by these N-hydroxyureas in the rat, it was considered that it precluded their development as therapeutic agents for use in man.
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  • 112
    ISSN: 1432-0738
    Keywords: Key words Cadmium ; Osteonectin ; Metallothionein ; Gene expression ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  Osteonectin gene expression in relation to metallothionein mRNA expression was investigated in various tissues from Cd-treated rats. After a single 50 μmol/kg subcutaneous injection of CdCl2, Cd predominantly accumulated in the liver and metallothionein gene expression significantly increased concomitantly with Cd accumulation, but no alteration of osteonectin gene expression was observed. In the kidney and lung, both metallothionein and osteonectin mRNA increased significantly but the elevation of metallothionein mRNA levels (1 h after Cd administration) preceded that of osteonectin (3 h after administration). A significant elevation of osteonectin mRNA levels was also observed in the testis after 3 h, but that of metallothionein mRNA occurred after 6 h. Not only accumulation of Cd but also increments in both osteonectin and metallothionein mRNA were minimal in the brain, but a significant increase in gene expression was observed after 1 h for osteonectin and after 3 h for metallothionein. Since, except in the testis, metallothionein gene expression preceded osteonectin gene expression, the induced metallothionein might transpose Cd and thereby affect its levels immediately, thus reducing the levels of Cd available for accumulation in other tissues. Hence, the osteonectin-Cd interaction might be secondary to the metallothionein-Cd interaction. However, the fact that osteonectin mRNA was predominantly induced by Cd administration in the target tissues of Cd toxicity, such as the lung, kidney and testis, suggests the possible involvement of osteonectin in Cd intoxication/detoxication mechanisms.
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  • 113
    ISSN: 1432-0738
    Keywords: Key words Paraquat ; Dinoseb ; 2 ; 4-D Intracellular calcium ; Rat ; Hepatocytes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  The effects of the herbicides paraquat, dinoseb and 2,4-D on intracellular Ca2+ levels and on vasopressin-induced Ca2+ mobilization were investigated in intact isolated hepatocytes. Incubation of rat hepatocytes with paraquat (5 mM for 60 min) and dinoseb (10 μM) resulted in a time-dependent loss of viability by approximately 25%. Viability of cells treated with 2,4-D decreased significantly, dropping to about 20% at 10 mM and 60 min incubation. Exposure of hepatocytes to paraquat (1–10 mM) for 60 min had no effect on the basal level of [Ca2+] i . Additionally, exposure to paraquat had no effect on the magnitude and on the duration of the [Ca2+] i response to vasopressin. In the presence of 2,4-D (1–10 mM), basal [Ca2+] i increases as a function of herbicide concentration. The magnitude of the Δ[Ca2+] i response decreases from 256±8 nM in control to 220±5 nM, at 10 mM 2,4-D. Exposure of hepatocytes to dinoseb (1–10 μM) had no effect on the basal level of [Ca2+] i . However, a strong concentration-dependent decrease in the magnitude of Δ[Ca2+] i in response to vasopressin was noticed at 60 min incubation. Dinoseb markedly inhibited the stimulation of the production of inositol phosphates by vasopressin stimulus. The present study demonstrates that paraquat, 2,4-D and dinoseb cause cell death in hepatocytes by mechanisms not related to an early increase in [Ca2+] i . Additionally, it has been shown for the first time that dinoseb disturbs the transduction mechanism promoted by vasopressin by inhibiting the formation of IP3.
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  • 114
    ISSN: 1432-0738
    Keywords: Key words β-Cyclodextrin ; Oncogenicity ; Rat ; Mouse
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  The results of oncogenicity studies of β-cyclodextrin in inbred Fischer 344 rats and CD-1 outbred mice are presented. Chronic feeding of β-cyclodextrin to Fischer 344 rats and CD-1 mice did not cause any treatment related carcinogenic effects. The only toxic effect was seen in mice as macroscopic distension of the large intestine with soft or fluid contents, histologically associated with the mucosa covered by mucous secretion containing exfoliated cells, and mucosal flattening and intestinal gland atrophy. Despite these observations, no differences between control and treated groups were observed concerning mortality, clinical observations or body weight and food consumption.
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  • 115
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    Archives of toxicology 70 (1995), S. 34-42 
    ISSN: 1432-0738
    Keywords: Key words Compartment model ; Erythrocytes ; Intracellular compartmentation ; Methyl mercury ; Rat ; Uptake
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  The uptake of methyl mercury (MeHg) by isolated rat erythrocytes was studied at 37°C using MeHg-cysteine (MeHgCySH), MeHg-glutathione (MeHgGSH), MeHg-mercaptalbumin (MeHgMASH) and the mixture of MeHgCySH with MeHgGSH, MeHgCySH with MeHgMASH, MeHgGSH with MeHgMASH at different MeHg concentrations. The measured MeHg concentrations were analyzed according to the Akaike’s information criterion in order to determine the suitable compartment model. After determining a two-compartment model, a model-independent two-compartment model was developed from the kinetics of uptake of MeHg at a concentration of 1 mmol MeHg/l packed erythrocytes using MeHgCySH, MeHgGSH and MeHgMASH, respectively. The developed two-compartment model was validated by predicting the kinetics of uptake of MeHg by rat erythrocytes at different MeHg concentrations and different mixtures of MeHg-complexes. Then, the predicted values were compared with the measured values. The results suggested: 1) MeHg uptake appeared suitable to be described by a two-compartment model, while using MeHgGSH, MeHgMASH, MeHgCySH at lower concentrations and the mixtures of MeHg-complexes; 2) MeHgCySH uptake was slowest among three kinds of MeHg-complexes, although a postulated cysteine-facilitated MeHgCySH transport system might exist in erythrocyte membrane; 3) the mixture of MeHg-complexes might facilitate MeHgCySH uptake; 4) there might be a second MeHg intracellular compartment in rat erythrocytes.
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  • 116
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    Archives of toxicology 69 (1995), S. 204-208 
    ISSN: 1432-0738
    Keywords: Key words Propyl gallate ; Rat ; Hepatocytes ; Cytotoxicity ; Gallate esters ; Antioxidant ; Mitochondria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  The cytotoxic effects of propyl gallate (PG), its related gallates and gallic acid have been studied in freshly isolated rat hepatocytes. Addition of PG (0.5–2.0 mM) to hepatocyte suspension elicited concentration-dependent cell death accompanied by losses of intracellular ATP, adenine nucleotide pools, glutathione (GSH) and protein thiols. The rapid loss of intracellular ATP preceded the onset of cell death caused by PG. In the comparative toxic effects of PG and related gallates at concentration of 1 mM, octyl gallate (OG), dodecyl gallate (DG) and butyl gallate (BG) elicited an abrupt depletion of ATP, followed by an acute cell death. These gallates were more toxic than PG; the toxic effects of PG were similar to those of methyl gallate (MG) and ethyl gallate (EG). In mitochondria isolated from rat liver, PG caused a concentration-dependent increase in the rate of state 4 oxygen consumption, indicating an uncoupling effect. The rate of state 3 oxygen consumption was inhibited by OG and DG. According to the respiratory control index, the order of impairment potency to mitochondria was OG〉BG, DG〉PG〉EG, MG〉gallic acid. These results indicate that PG and related gallates are toxic to hepatocytes and that the acute cytotoxicity may be due to mitochondrial dysfunction.
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  • 117
    ISSN: 1432-0738
    Keywords: Key words Eosinophilia-myalgia syndrome ; L-Tryptophan ; 3-(Phenylamino) alanine ; Rat ; 1 ; 1’-Ethylidenebis (L-tryptophan)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  Consumption of certain product lots of L-tryptophan (LT) has been reported to be epidemiologically associated with an outbreak of eosinophilia-myalgia syndrome (EMS) in the United States. Since the production lots were found to contain 3-phenylamino alanine (PAA) as an impurity, its effects were studied by administering the substance orally by gavage to 5-week-old Sprague-Dawley rats. Groups of animals were given PAA for 13 consecutive weeks at dose levels of 1, 10 and 100 mg/kg per day. The animals were killed at 4 or 8 weeks. Hematological and blood biochemical tests were performed and detailed histopathological observations were made. No significant abnormalities were observed in the test animals and in particular no EMS-like conditions. A brief summary of other animal studies using several species of rats and mice performed in our laboratory since 1989 on various LT related substances is also presented. No EMS-like effects were observed in these studies.
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  • 118
    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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  • 119
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    Psychopharmacology 117 (1995), S. 154-161 
    ISSN: 1432-2072
    Keywords: Antidepressants ; DRL 72s schedule Water reinforcement ; Chlordiazepoxided-Amphetamine ; Haloperidol ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The effects of three antidepressants, desipramine (2.5–20 mg/kg) tranylcypromine (0.63–2.5 mg/kg) mianserin (1.25–10 mg/kg) and three non-antidepressants, chlordiazepoxide (CDP; 1.25–10 mg/kg) haloperidol (0.02–0.16 mg/kg)d-amphetamine (0.31–1.25 mg/kg) were evaluated in rats responding for water reinforcement under a DRL 72s schedule. The antidepressants all produced dose-related decreases in overall response rates, but no significant changes in reinforcement frequency. In contrast, the anxiolytic CDP did increase the number of reinforcers obtained. Haloperidol decreased both reinforcers and responses whilstd-amphetamine stimulated responding, thereby decreasing reinforcement frequency. An analysis of the modes of inter-response times (IRTs) revealed no significant shifts in the peaks of the IRT distributions for most of the drugs tested. Amphetamine, however, (0.31 and 0.63 mg/kg) decreased the modal values in correspondence with the shift to the left of the peak of responding caused by this compound. These results are discussed in the context of the use of the DRL 72s procedure as a screening test for antidepressant drugs.
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  • 120
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    Psychopharmacology 117 (1995), S. 240-247 
    ISSN: 1432-2072
    Keywords: Microdialysis Conditioned place preference ; Morphine Locomotor activity ; Genetic differences ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Studies examining differential sensitivity to psychoactive drugs in mice suggest that genotype may play a critical role. Furthermore, an involvement of genotype in mediating individual differences in sensitivity to the rewarding effects of several drugs of abuse has also been postulated. The aim of this study was to examine the conditioned rewarding and dopamine-releasing effects of morphine in two outbred rat strains commonly used in addiction research. Additionally, the behavioural and neuroendocrine responses of these strains to the stress of novelty were also examined. Basal locomotor activity was higher in Wistar rats than Sprague-Dawley following exposure to a novel environment. In contrast, elevations in plasma corticosteroid levels following novelty exposure did not differ between the two strains. In a counterbalanced place preference conditioning procedure, increasing doses of morphine (1.0–10.0 mg/kg SC) produced significant conditioned place preferences (CPP) in both Wistar and Sprague-Dawley strains. However, Wistar rats required a significantly larger dose of morphine (5.0 mg/kg) to produce a significant CPP than the Sprague-Dawley rats. In the latter strain, CPP occurred with doses of 3.0 mg/kg and greater. In parallel microdialysis experiments, both strains showed significant dose-related increases in dopamine release in the nucleus accumbens following acute morphine challenge (1.0–10.0 mg/kg SC). Again in Wistar rats, a larger dose of morphine was necessary to produce a significant increase in comparison to Sprague-Dawley rats. These results show that genetically distinct rat strains can show differential sensitivity to opioids, more specifically to drug-seeking responses.
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  • 121
    ISSN: 1432-2072
    Keywords: Microdialysis ; Dorsal hippocampus 5-HT1A and 5-HT1B receptors ; Chronic treatment Fluvoxamine ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract This study investigated the alterations of the 5-HT1A and 5-HT1B autoreceptor function following chronic treatment with fluvoxamine using osmotic minipumps. The 5-HT1A and 5-HT1B autoreceptor function were studied using microdialysis in the dorsal hippocampus. The effect of the 5-HT1A receptor agonist 8-OH-DPAT (0.3 mg/kg, SC) and the 5-HT1B receptor agonist RU-24969 (100 nM through the dialysis probe for 30 min) on 5-HT release was compared with rats chronically treated with saline. 8-OH-DPAT decreased 5-HT release to 55% and 60% of baseline, while RU-24969 decreased 5-HT release to 66% and 70% of baseline value in the saline and fluvoxamine group, respectively. In both cases, differences between the saline and fluvoxamine groups were not statistically significant. Plasma levels of fluvoxamine after 21 days of treatment ranged from 3 to 5 ng/ml. Fluvoxamine concentration in rat brain during treatment was estimated between 100 and 200 nM, which approximates to the IC50 value of fluvoxamine on the 5-HT transporter in synaptosomes and is 50 times higher than the Kd value for the 5-HT reuptake site. In conclusion, no evidence was found for changes in 5-HT1A,B receptor function using 8-OH-DPAT and RU-24969 as probes after continuous treatment with fluvoxamine by means of osmotic minipumps.
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  • 122
    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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  • 123
    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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  • 124
    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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  • 125
    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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  • 126
    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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  • 127
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    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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  • 128
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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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  • 129
    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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  • 130
    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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  • 131
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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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  • 132
    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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  • 133
    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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  • 134
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    Psychopharmacology 118 (1995), S. 226-229 
    ISSN: 1432-2072
    Keywords: Touch-sensitive device ; Visual attention ; Cognition ; Amphetamine ; Serotonin ; 5-HT ; Acetylcholine ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract A multiple choice serial reaction time task was used to investigate visual attention in rats. The intelligence panel consisted of a transparent touch-sensitive device, placed directly in front of a video monitor. Amphetamine (0.2–1.6 mg/kg IP) increased errors of omission and decreased latency to respond, but had no effect on accuracy. The 5-HT agonist quipazine (0.6–2.4 mg/kg IP) increased errors of omission, but did not affect other parameters. ICV administration of hemicholinium-3 (1–4 µg) had no effect upon performance. Thus, psychopharmacological manipulations can reliably alter performance in the touch window box, suggesting potential new avenues for rat cognitive testing.
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  • 135
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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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  • 136
    ISSN: 1432-2072
    Keywords: Phencyclidine ; Dorsal hippocampus ; Rat ; Spatial navigation task
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Since the hippocampus is likely to be a major site of phencyclidine (PCP) action, the effects of various doses of PCP (1.8, 18 or 36 nM) as well as 3.6 nM MK-801 or saline injected directly into the dentate gyrus of the hippocampus was tested for acquisition of a spatial navigation task (dry land version of a water maze) using a paradigm that assesses short term memory based on learning within a day and long term memory based on learning between days. Results indicated that relative to saline or 1.8 nM PCP injected rats, rats with 18 or 36 nM PCP or 3.6 nM MK-801 injections were impaired in acquisition of the task as measured by increased distances traveled to find the food location between days but not within days. In additional experiments 36 nM PCP or 3.6 nM MK-801 did not produce any deficits in the acquisition of an object discrimination task. It is suggested that PCP through its blocking action of the NMDA receptor in the dentate gyrus or CA1 region of the dorsal hippocampus mediates the consolidation of new spatial location information.
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  • 137
    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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  • 138
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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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  • 139
    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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  • 140
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    Psychopharmacology 122 (1995), S. 1-14 
    ISSN: 1432-2072
    Keywords: Rat ; Development ; Behavior ; Cannabis ; Delta-9-tetrahydrocannabinol ; Motor activity ; Place preference ; Grooming ; Corticosterone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Cannabis sativa preparations (hashish, marijuana) are the most widely used illicit drugs during pregnancy in Western countries. The possible long-term consequences for the child of in utero exposure to cannabis derivatives are still poorly understood. Animal models of perinatal cannabinoid exposure provide a useful tool for examining the developmental effects of cannabinoids. Behavioral consequences of maternal exposure to either cannabis preparations or to its main psychoactive component, Δ9-tetrahydrocannabinol (THC) in rat models are reviewed in this paper. Maternal exposure to cannabinoids resulted in alteration in the pattern of ontogeny of spontaneous locomotor and exploratory behavior in the offspring. Adult animals exposed during gestational and lactational periods exhibited persistent alterations in the behavioral response to novelty, social interactions, sexual orientation and sexual behavior. They also showed a lack of habituation and reactivity to different illumination conditions. Adult offspring of both sexes also displayed a characteristic increase in spontaneous and water-induced grooming behavior. Some of the effects were dependent on the sex of the animals being studied, and the dose of cannabinoid administered to the mother during gestational and lactational periods. Maternal exposure to low doses of THC sensitized the adult offspring of both sexes to the reinforcing effects of morphine, as measured in a conditioned place preference paradigm. The existence of sexual dimorphisms on the developmental effects of cannabinoids, the role of sex steroids, glucocorticoids, and pituitary hormones, the possible participation of cortical projecting monoaminergic systems, and the mediation of the recently described cannabinoid receptors are also analyzed. The information obtained in animal studies is compared to the few data available on the long-term behavioral and cognitive effects on in utero exposure to cannabis in humans.
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  • 141
    ISSN: 1432-2072
    Keywords: Prepulse inhibition ; Schizophrenia ; Dopamine ; Serotonin ; Glutamate ; Social isolation ; Clozapine ; Raclopride ; Haloperidol ; Rirsperidone ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Prepulse inhibition (PPI) of an acoustic startle response is impaired in schizophrenics. PPI can also be studied in the rat, and is impaired by dopamine (DA) D2/3 receptor agonists such as apomorphine. This disruption is reversed by DA antagonists, leading to proposals that this approach may be a useful means to identify novel antipsychotics. There is also evidence to suggest a role of serotonergic (5-HT) and glutamatergic systems in schizophrenia, and accordingly PPI can be disrupted by the 5-HT2 agonist DOI, and the non-competitive NMDA antagonist, dizocilpine. In the present study we have examined the effect of four antipsychotic drugs, haloperidol (0.1–0.3 mg/kg), raclopride (0.03–0.3 mg/kg), risperidone (0.3–3 mg/kg) and clozapine (0.0001–10 mg/kg), against the PPI disruptions induced by apomorphine (0.5 mg/kg), DOI (3 mg/kg) and dizocilpine (0.15 mg/kg). Furthermore, these drugs have been examined for their ability to restore a PPI deficit produced by housing rats under conditions of social isolation. All drugs except clozapine reversed an apomorphine-induced disruption. However, clozapine and risperidone, but not raclopride and haloperidol, reversed a DOI-induced disruption. Only risperidone was effective in restoring a PPI deficit produced by dizocilpine. In contrast to the drug-induced disruptions which were differentially sensitive to the various neuroleptics, isolation-induced disruptions were restored by each drug. These results support the idea that non-drug induced disruptions of PPI, such as social isolation, may be a more viable approach to identify novel antipsychotics.
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  • 142
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    Virchows Archiv 427 (1995), S. 181-186 
    ISSN: 1432-2307
    Keywords: Isoproterenol ; Apoptosis ; Rat ; Parotid ; Sialadenosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Enlargement of the rat parotid salivary glands was induced by repeated administration of isoproterenol. Mean wet weights of the treated glands increased steadily to 240% of control values. Following withdrawal of the drug, quantitative histological techniques were used to investigate the balance between hypertrophy, hyperplasia and apoptosis. The volume occupied by acinar cells relative to the total gland volume together with cytoplasmic|:|nuclear area ratios as measures of hypertrophy increased during the early experimental period. Similarly, serous acinar cell mitotic counts increased, indicating that hyperplasia had occurred. Apoptosis was demonstrated at light microscopical level to be the main mechanism for cell deletion as the glands returned to normal size and weight. The results indicate that hypertrophy and hyperplasia of serous acinar cells contribute to isoproterenol-induced sialadenosis. The experimental animal model demonstrates that these proliferative changes are completed by 48 h and thereafter are balanced by apoptosis as the glands recover their normal size and weight.
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  • 143
    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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  • 144
    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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  • 145
    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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  • 146
    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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  • 147
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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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  • 148
    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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  • 149
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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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  • 150
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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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  • 151
    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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  • 152
    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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  • 153
    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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  • 154
    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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  • 155
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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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  • 156
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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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  • 157
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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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  • 158
    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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  • 159
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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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  • 160
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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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  • 161
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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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  • 162
    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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  • 163
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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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  • 164
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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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  • 165
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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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  • 166
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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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  • 167
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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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  • 168
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    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.
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  • 169
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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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  • 170
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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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  • 171
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    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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  • 172
    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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  • 173
    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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  • 174
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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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  • 175
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    New York, NY [u.a.] : Wiley-Blackwell
    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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  • 176
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    New York, NY [u.a.] : Wiley-Blackwell
    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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  • 177
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    New York, NY [u.a.] : Wiley-Blackwell
    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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