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Notes: Abstract: Slices of rabbit caudate and hypothalamus take up and accumulate [3H]imipramine. In superfused slices of both structures electrical stimulation or exposure to tyramine failed to release recently taken up [3H]imipramine. De-polarization by exposure to 30–60 mm-potassium caused only a small release of [3H]imipramine that was not concentration-dependent. The release of [3H]imipramine by high potassium was independent of the presence of calcium ions in the superfusion medium. These results contrasted with those obtained for the release of [3H]dopamine from the caudate and [3H]noradrenaline from the hypothalamus, where tyramine, electrical stimulation, and high potassium caused a significant release of the labeled neurotransmitters. The release of [3H]dopamine from the caudate and [3H]noradrenaline from the hypothalamus elicited by electrical stimulation or high potassium was entirely calcium-dependent. It is concluded that [3H]imipramine is taken up into the two brain regions and is accumulated in a nonvesicular site from which it is not released by calcium-dependent depolarizing stimuli.

Notes: Abstract: High-affinity specific [3H]imipramine binding has been demonstrated in the brain and platelets of various species including man. Electrolytic lesion of the rat dorsal raphe, which resulted in a significant decrease in the endogenous levels of serotonin produced a reduction in the density of [3H]imipramine binding sites in the hypothalamus and cortex. The affinity constants were unchanged. These results suggest that [3H]imipramine binding sites are located on serotonin nerve terminals.

Notes: Abstract: 5-Methoxytryptoline potently inhibits [3H]-imipramine binding to membranes from the cerebral cortex and platelets. Since 5-methoxytryptoline, which appears to occur endogenously with particularly high levels in the human pineal gland, also inhibits 5-hydroxy-tryptamine (5-HT, serotonin) uptake, it should be considered as a putative endogenous ligand modulating 5-HT transport. As the 5-HT transporter complex comprises the imipramine and the substrate recognition sites, which interact allosterically, it was essential to define the mechanism of inhibition of [3H]imipramine binding by 5-methoxytryptoline. Human platelets show an active and saturable uptake of 5-HT and tryptamine. The uptake of both substrates appears to be mediated by the same carrier and it is inhibited by 5-methoxytryptoline at submicromolar concentrations. 5-HT and tryptamine inhibit [3H]imipramine binding in human platelets with a Hill slope for inhibition close to unity and IC70 values of 3,265 and 3,475 nM, respectively. This inhibition is, however, not competitive because both 5-HT and tryptamine significantly decrease the rate of [3H]imipramine-receptor dissociation. Although 5-methoxytryptoline potently inhibits [3H]imipramine binding (IC50= 44 nM) in human platelets with a Hill slope of unity, it does not affect the receptor-ligand dissociation rate of [3H]imipramine even at concentrations up to 100 μM. The present experiments show that 5-methoxytryptoline, in spite of its chemical similarity to the indoleamine transporter substrates, interacts with the imipramine receptor through a mechanism of competitive inhibition. This conclusion is supported by a selective effect of 5-methoxytryptoline on the Kd of [3H]imipramine binding. These data are compatible with the hypothesis that 5-methoxytryptoline may be an endogenous modulator that interacts competitively with the imipramine receptor associated with the 5-HT uptake complex.

Notes: Abstract: [3H]Imipramine binds with high affinity to membranes from different regions of the human brain. The highest density of binding sites was observed in the hypothalamus and substantia nigra and the lowest density in the white matter and cerebellum. As found in rat brain, tricyclic antidepressant drugs are potent inhibitors of [3H]imipramine binding. Atypical antidepressants are, however, much weaker at inhibiting the specific binding. The [3H]imipramine binding site in human cortex is apparently identical to the site already described in the rat brain and in human platelets.

Notes: Abstract: In the rat brain, the presynaptic 5-hydroxytrypt-amine (5-HT) autoreceptors located on 5-HT terminals correspond to the 5-HT1B subtype. The presence of a 5-HT receptor probably located on 5-HT nerve endings and modulating transmitter release in the human neocortex has been reported, but its detailed pharmacological characterization is not yet available. On the other hand, receptor binding and autoradiographic results indicate that the 5-HT1B receptor subtype is not present in the human brain. We, therefore, studied the modulation of the electrically evoked release of [3H]5-HT by various 5-HT receptor agonists and antagonists in preloaded slices of human neocortex obtained from 18 patients undergoing neurosurgery. The nonselective 5-HT1A/1B/1D receptor agonist 5-carboxamidotryptamine produced a potent inhibition (70% at 0.03 μM) of the electrically evoked release of [3H]5-HT which was blocked by 5-HT receptor antagonists with the following relative order of potency: methiothepin 〉 metergoline = methysergide 〉 propranolol. The selective 5-HT1A receptor agonist 8-hydroxy-2–(di-n-propylamino)tetralin at 0.1 μM did not modify the electrically evoked release of [3H]5-HT. The 5-MT1A/1B receptor agonist RU 24969 was 10 times more potent at inhibiting [3H]5-HT overflow in the rat frontal cortex than in the human neocortex. The potent 5-HT1B receptor antagonist cyanopindolol did not modify the 5-carboxamidotryptamine-induced inhibition of the electrically evoked release of [3H]5-HT in slices of the human neocortex, but produced by itself a small inhibition of [3H]5-HT overflow. The α2-adrenoceptor antagonist yohimbine, which possesses affinity for the 5-HT1D receptor subtype, decreased the release of [3H]5-HT, but only in the presence of the selective α2-adrenoceptor antagonist idazoxan, which by itself increased significantly [3H]5-HT overflow. Taken together, these results support the view that the 5-HT receptor modulating the electrically evoked release of [3H]5-HT in slices of the human neocortex could be of the 5-HT1D subtype. Moreover, preliminary results obtained with idazoxan confirm the existence of a presynaptic α2-adrenoceptor modulating the release of [3H]5-HT in the human neocortex. These α2-heteroreceptors could exert a tonic inhibitory modulation on 5-HT neurotransmission in the human neocortex.

Notes: Abstract: The regional distribution of [3H]zoIpidem, a novel imidazopyridine hypnotic possessing preferential affinity for the BZD1 (benzodiazepine subtype 1) receptor, has been studied autoradiographically in the rat CNS and compared with that of [3H]flunitrazepam. The binding of [3H]zolpidem to rat brain sections was saturable, specific, reversible, and of high affinity (KD= 6.4 nM). It occurred at a single population of sites whose pharmacological characteristics were similar to those of the benzodiazepine receptors labeled with [3H]flunitrazepam. However, ethyl-β-carboline-3-carboxylate and CL 218,872 were more potent displacers of [3H]zolpidem than of [3H]flunitrazepam. The autoradiographic brain distribution of [3H]zolpidem binding sites was qualitatively similar to that previously reported for benzodiazepine receptors. The highest levels of [3H]- zolpidem binding sites occurred in the olfactory bulb (glomerular layer), inferior colliculus, ventral pallidum, nucleus of the diagonal band of Broca, cerebral cortex (layer IV), medial septum, islands of Calleja, subthalamic nucleus, and substantia nigra pars reticulata, whereas the lowest densities were found in parts of the thalamus, pons, and medulla. Comparative quantitative autoradiographic analysis of the binding of [3H]zolpidem and [3H]flunitrazepam [a mixed BZD1/BZD2 (benzodiazepine subtype 2) receptor agonist] in the CNS revealed that the relative density of both 3H- labeled ligands differed in several brain areas. Similar levels of binding for both ligands were found in brain regions enriched in BZD1 receptors, e.g., substantia nigra pars reticulata, inferior colliculus, cerebellum, and cerebral cortex lamina IV. The levels of [3H]zolpidem binding were five times lower than those of [3H]flunitrazepam binding in those brain regions enriched in BZD2 receptors, e.g., nucleus accumbens, dentate gyrus, and striatum. Moreover. [3H]zolpidem binding was undetectable in the spinal cord (which contains predominantly BZD2 receptors). Finally, like CL 218,872 and ethyl-β-carboline-3-carboxylate, zolpidem was a more potent displacer of [3H]flunitrazepam binding in brain regions enriched in BZD1 receptors than in brain areas enriched in BZD2 receptors. The present data add further support to the view that zolpidem, although structurally unrelated to the benzodiazepines, binds to the benzodiazepine receptor and possesses selectivity for the BZD1 receptor subtype.

Notes: Abstract: The activation by endogenous dopamine of the inhibitory 3,4-dihydroxyphenylethylamine (dopamine) receptors modulating the electrically evoked release of [3H]acetylcholine ([3H]ACh) and [3H]dopamine in rat striatal slices is a function of the concentration of dopamine accumulated in the synaptic cleft during electrical stimulation. When the release of 3H-neurotransmitters was elicited with a 2-min period of stimulation at a frequency of 1 Hz, neither dopamine autoreceptors nor dopamine receptors modulating [3H]ACh were activated by endogenously released dopamine. On the other hand, exposure to (S)-sulpiride facilitated the release of [3H]dopamine and [3H]ACh elicited when the 2-min stimulation was carried out at a frequency of 3 Hz but this effect was not observed at a lower frequency of stimulation (1 Hz). In the presence of amphetamine the dopamine receptors modulating the electrically evoked release of [3H]ACh can be activated by endogenous dopamine even at the lower frequency of stimulation (1 Hz). Similar effects can be obtained if the neuronal uptake of dopamine is inhibited by cocaine or nomifensine. The inhibition by amphetamine of the release of [3H]ACh elicited by electrical stimulation at 1 Hz involves dopamine receptors and can be fully antagonized by clozapine, haloperidol, chlorpromazine, or pimozide. The stereoselectivity of this antagonism can be demonstrated with the optical enantiomers of sulpiride and butaclamol. This inhibitory effect of amphetamine on cholinergic neurotransmission appears to be the result of the stimulation of dopamine receptors of the D2 subtype, as they were resistant to blockade by the preferential D1 receptor antagonist SCH 23390. Inhibition of dopamine synthesis with α-methyl-p-tyrosine antagonized the effects of amphetamine on [3H]ACh release. The dopamine receptor-mediated inhibition of the release of [3H]ACh elicited at low frequencies of stimulation from rat striatal slices is a suitable model to test drugs that enhance dopaminergic neurotransmission. In addition, the antagonism of the inhibition of cholinergic neurotransmission by amphetamine in striatal slices represents a useful test for drugs with potential usefulness in clinical situations in which dopaminergic neurotransmission is exacerbated.

Notes: Abstract: Tricyclic antidepressants and nontricyclic serotonin (5-hydroxytryptamine) uptake blockers monophasically inhibit [3H]imipramine binding in human platelets. Similarly, serotonin and tryptamine inhibit the binding of [3H]imipramine in the low micromolar range and with a pseudo-Hill coefficient near unity. Dissociation of the [3H]imipramine receptor complex in the presence of uptake inhibitors follows first-order kinetics with a half-life of approximately 60 min. Although serotonin and tryptamine do not decrease [3H]imipramine binding when added under equilibrium conditions, simultaneous addition of serotonin or tryptamine with serotonin uptake inhibitors decreases the rate of ligand-receptor dissociation in a concentration-dependent manner. These data suggest a common site of action for serotonin, which is the substrate of the transporter system, and of tryptamine, its nonhydroxylated analog. This hypothesis is supported by the identification of a high-affinity (Km= 0.55 μM), saturable, and temperature-dependent uptake of [3H]tryptamine in human platelets. Uptake of [3H]tryptamine was inhibited potently by imipramine and nontricyclic serotonin uptake inhibitors with a potency similar to that observed for [3H]serotonin uptake. These data support the hypothesis that in platelets, [3H]imipramine, tricyclic, and nontricyclic serotonin up-take inhibitors bind to a common recognition site that is associated with the serotonin transporter but that differs from the substrate recognition site of the carrier through which serotonin and tryptamine exert a heterotropic allosteric modulation on [3H]imipramine binding.

Notes: Aims : To determine the pattern of macrophage infiltration in colon cancers and its correlation with clinicopathological characteristics.Methods and results : Colon cancers from 100 patients were arrayed into a tissue microarray (TMA). Four cores per tumour were taken: three from the invasion front (IF) and one from the tumour surface (TS). Macrophages were quantified by immunohistochemistry with antibodies to the PG-M1, KP-1, MRP8, MRP14 and MRP8/14 antigens. The number of macrophages was significantly higher in the TS cores than in the IF cores and both tumour sites showed a higher number of macrophages than the normal mucosa. The number of macrophages decreased in higher stage tumours. The different tumour-associated macrophage (TAM) subpopulations were positively correlated with each other.Conclusions : The increased number of macrophages in cancers compared with normal colon mucosa indicates that macrophages are attracted to the tumour site. However, decreasing macrophages in higher stage colon cancers suggest that this attraction decreases with tumour progression.

Notes: In this paper we report the results of experiments that compare the x-ray emission from a laser spot in a radiation-filled hohlraum to that from a similar laser spot on a simple disk target. The studies were done using the Nova laser facility [J. D. Lindl, Phys. Plasmas 2, 3933 (1995)] in its 0.35 μm wavelength, 1 ns square pulse configuration. Focal spot intensities were 2–3.5×1015 W/cm2. X-ray images measured x-ray conversion in a hohlraum and from an isolated disk simultaneously. A laser spot inside a hohlraum emitted more x rays, after subtracting the background emission from the hohlraum walls, than a spot on a disk. Numerical models suggest the enhanced spot emission inside the hohlraum is due to an increase in lateral transport relative to the disk. Filamentation in the hohlraum will also increase the spot size. The models agree fairly well with the results on spot spreading but do not explain the overall increase in conversion efficiency. © 1997 American Institute of Physics.