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A Beaudet

Publications and source records attributed to A Beaudet.

At least 109 records · Page 6Linked to original sources

Expression of a marrow stroma and thymus-associated antigen (ST3) in the rat brain: comparison with Thy-1.

Cells of the immunohemopoietic and nervous systems express certain molecules that generally are not found in other tissues. One example is the 'ST3' antigen, which is present on the major population of fibroblastoid cells grown from rat bone marrow, but is not detected on adherent cells from most peripheral organs (e.g. lung). An immunohistological survey revealed ST3 also in the thymic cortex, the glomerular mesangial area, and the brain. Because this pattern of distribution is similar to that described for Thy-1, we compared the localization of the two antigens in the adult rat brain and found that there were areas where it was congruent and others where it was distinct. Staining for ST3 was absent from the white matter, but was especially notable in discrete layers of the frontal, orbital, parietal, and cingulate cortices, the substantia nigra, the inferior olivary nuclei, and the deep molecular layer of the cerebellum, as well as other scattered regions in the gray matter. This is in contrast to Thy-1, which stained more diffusely throughout the gray zones. In further experiments using primary brain cell cultures, ST3 was demonstrated on neurons, but not on oligodendrocytes or astrocytes. Similarly, it was found on the surface of cells of the PC12 neuronal line, but not on the C6 astrocytoma. This restricted distribution on a subpopulation of neurons raises the possibility that the ST3 epitope might be part of a cell interaction molecule of the marrow stroma, thymus, and brain.

Animals↗

Immunoautoradiographic localization of interleukin 2-like immunoreactivity and interleukin 2 receptors (Tac antigen-like immunoreactivity) in the rat brain.

Immunoautoradiographic techniques were used to determine the topographical distribution of interleukin 2-like immunoreactivity and interleukin 2 receptor-like immunoreactivity (Tac antigen-like immunoreactivity) in rat brain. Interleukin 2 receptors were also visualized by film autoradiography using 125I-recombinant human interleukin 2. Endogenous interleukin 2-like immunoreactive material was present in a limited number of brain regions. The highest densities were localized to the median eminence-arcuate nucleus complex, hippocampal formation, lamina IV of the cerebral cortex, lateral septum, neostriatum and cerebellum. Lower levels of interleukin 2-like immunoreactive material were present in the thalamus, medial septum and granule cell layer of the cerebellar cortex. Tac antigen-like immunoreactivity was observed in virtually the same brain regions, and within these brain regions showed the same distribution as interleukin 2-like immunoreactivity. In contrast, [125I]interleukin 2 binding sites were only detected in the hippocampal formation and the molecular layer of the cerebellar cortex. Quantitative analyses confirmed that there was a positive correlation between the densities of interleukin 2-like immunoreactivity and interleukin 2 receptor immunoreactivity (Tac antigen-like immunoreactivity) in various brain regions, suggesting that interleukin 2 is synthesized and/or stored in the vicinity of the site of interaction with the Tac antigen of its receptor. Overall, the presence of endogenous interleukin 2-like immunoreactive material and interleukin 2 receptor-like immunoreactive material in selective regions of the rat brain suggests that this neurokine may normally act to regulate a variety of brain functions in the adult rat.

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High resolution radioautographic localization of [125I]FK-33-824-labelled mu opioid receptors in the spinal cord of normal and deafferented rats.

Recent data have shown that [125I]D-Ala2, MePhe4, Met(o)ol5-enkephalin (FK-33-824) is a highly selective and specific mu opioid receptor ligand [Moyse et al. (1986) Peptides 7, 351-355]. This probe was used here to investigate the detailed radioautographic distribution of mu sites at various levels of the spinal cord. [125I]FK-33-824 binding sites were localized by both tritium-sensitive film and liquid emulsion radioautography in the spinal cord of naive and deafferented rats. In naive animals, high densities of mu sites were apparent within laminae I-II at all levels of the dorsal horn, with higher levels of labelling seen in layer IIi as compared to IIo in the lumbar segment. Laminae III-IV contained about half the quantities of binding observed in superficial layers. Relatively high densities of sites were also seen over lamina VI in the upper cervical cord and throughout Clarke's column. Within the latter, [125I]FK-33-824 binding clearly spared the large perikarya of the spinocerebellar neurons. In the ventral horn, [125I]FK-33-824 binding was mainly concentrated in layer IX, at the level of cervical and lumbar enlargements. Labelled sites were confined to the neuropil, mostly sparing the soma of motoneurons. Significant decreases in [125I]FK-33-824 binding in laminae I-II (55%) and III-IV (28%) were detected four days following cervical (C3-C7) or lumbar (L1-L6) rhizotomies. These decrements were most evident at seven days post-lesion at C3-C7 levels (93 and 76% in laminae I-II and III-IV, respectively) and recovered slightly thereafter up to 28 days post-lesion. In contrast, dorsal rhizotomies did not influence mu labelling in either the ventral horn or Clarke's column. These results confirm the association of mu opioid binding sites with dorsal primary afferent fibres and demonstrate the presence of mu sites in Clarke's column and lamina IX of the ventral horn. These findings suggest that endogenous opioids in the spinal cord play a role in sensory motor integration as well as in the modulation of primary nociceptive inputs.

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Neurotensin regulation of endogenous acetylcholine release from rat striatal slices is independent of dopaminergic tone.

The effects of neurotensin (NT) alone or in combination with the dopamine antagonist sulpiride were tested on the release of endogenous acetylcholine (ACh) from striatal slices. NT enhanced potassium (25 mM)-evoked ACh release from striatal slices in a dose-dependent manner. This effect was tetrodotoxin-insensitive, suggesting an action directly on cholinergic elements. The dopamine antagonist sulpiride (5 x 10(-5) M) significantly increased (63%) potassium-evoked ACh release from striatal slices; potassium-evoked ACh release was further increased (90%) in the presence of NT (10(-5) M) and sulpiride (5 x 10(-5) M). The second set of experiments tested the effects of 6-hydroxydopamine (6-OHDA) lesions of the substantia nigra on NT-induced increases of potassium-evoked ACh release. These lesions did not alter the NT regulation of potassium-evoked ACh release from striatal slices, but did significantly increase spontaneous (33%) and potassium-evoked (40%) ACh release from striatal slices. Striatal choline acetyltransferase activity was not affected by 6-OHDA lesions. In addition, following 6-OHDA lesions, sulpiride was ineffective in altering ACh release from striatal slices. Furthermore, evoked ACh release in the presence of the combination of NT and sulpiride was not different from that in the presence of NT alone. These results suggest that in the rat striatum, NT regulates cholinergic interneuron activity by interacting with NT receptors associated with cholinergic elements. Moreover, the NT modulation of cholinergic activity is independent of either an interaction of NT with D2 dopamine receptors or the sustained release of dopamine.

Acetylcholine↗

Distribution of mu, delta, and kappa opioid receptors in the hypothalamus of the rat.

The radioautographic distribution of mu, delta and kappa opioid binding sites was examined by in vitro radioautography in the rat hypothalamus using the highly selective ligands [125I]-FK 33-824, [125I]azidoDTLET and [125I]DPDYN, respectively. Levels of mu opioid binding sites varied considerably amongst hypothalamic nuclei. mu Opioid labeling was dense in the medial preoptic area, medial preoptic nucleus, suprachiasmatic nucleus and ventromedial nucleus, whilst the supraoptic nucleus, paraventricular nucleus, arcuate nucleus and dorsomedial nucleus were devoid of labeling. Delta opioid labeling was sparse throughout most of the hypothalamus; however, moderate binding densities were detected in the suprachiasmatic and ventromedial nucleus. kappa Opioid labeling was also scant throughout the hypothalamus with the exception of the suprachiasmatic nucleus which was very densely labeled. Our results indicate that the 3 opioid receptors types are differentially distributed within the hypothalamus, although a significant overlap exists. In general, the distribution of hypothalamic opioid receptors correlates well with that of opioid-containing terminal fibers and may represent the anatomical substrate for opioid involvement in the hypothalamic regulation of autonomic, behavioral and neuroendocrine functions.

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Chronic estradiol treatment alters central cholinergic function in the female rat: effect on choline acetyltransferase activity, acetylcholine content, and nicotinic autoreceptor function.

The purpose of the present study was to determine the effects of chronic (2-month) estradiol (E2) treatment on cholinergic function in the frontal cortex, hippocampus and hypothalamus of the female rat. Chronic E2 treatment selectively increased choline acetyltransferase (ChAT) activity in the frontal cortex and decreased ChAT activity in the hypothalamus: hippocampal ChAT activity was not significantly changed. The decrease of ChAT activity in the hypothalamus was paralleled by a significant reduction in the content and release (basal and evoked) of acetylcholine (ACh) in this structure. Neither the content nor the release of ACh was altered in the frontal cortex. In the hippocampus, there was a significant increase in spontaneous ACh release; however, hippocampal ACh content and evoked ACh release were not changed. Chronic E2 treatment also altered nicotinic binding sites in these same regions as determined by saturation analysis of [3H]methylcarbamylcholine (MCC) to membranes. There was a decrease in the affinity and an increase in the density of [3H]MCC binding sites in hippocampal membranes and an increase in the density of [3H]MCC binding sites was observed in hypothalamic membranes. These alterations were paralleled by changes in nicotinic autoreceptor function within these two structures. In the hippocampus, the increase in spontaneous ACh release induced by MCC in control animals was no longer apparent after chronic treatment with E2. In hypothalamus, higher concentrations of the nicotinic agonist were required to increase spontaneous ACh release in slices from E2-treated rats as compared to control rats. Taken together, these results suggest that chronic E2 treatment decreases presynaptic cholinergic function in the female rat hippocampus and hypothalamus.

Acetylcholine↗

Distribution of 125I-neurotensin binding sites in human forebrain: comparison with the localization of acetylcholinesterase.

The distribution of 125I-neurotensin binding sites was compared with that of acetylcholinesterase reactivity in the human basal forebrain by using combined light microscopic radioautography/histochemistry. High 125I-neurotensin binding densities were observed in the bed nucleus of the stria terminalis, islands of Calleja, claustrum, olfactory tubercle, and central nucleus of the amygdala; lower levels were seen in the caudate, putamen, medial septum, diagonal band nucleus, and nucleus basalis of Meynert. Adjacent sections processed for cholinesterase histochemistry demonstrated a regional overlap between the distribution of labeled neurotensin binding sites and that of intense acetylcholinesterase staining in all of the above regions, except in the bed nucleus of the stria terminalis, claustrum, and central amygdaloid nucleus, where dense 125I-neurotensin labeling was detected over areas containing only weak to moderate cholinesterase staining. At higher magnification, 125I-neurotensin-labeled binding sites in the islands of Calleja, supraoptic nucleus of the hypothalamus, medial septum, diagonal band nucleus, and nucleus basalis of Meynert were selectively associated with neuronal perikarya found to be cholinesterase-positive in adjacent sections. Moderate 125I-neurotensin binding was also apparent over the cholinesterase-reactive neuropil of these latter three regions. These data suggest that neurotensin (NT) may directly influence the activity of magnocellular cholinergic neurons in the human basal forebrain, and may be involved in the physiopathology of dementing disorders such as Alzheimer's disease, in which these neurons have been shown to be affected.

Acetylcholinesterase↗

Ultrastructural localization of [125I]neurotensin binding sites to cholinergic neurons of the rat nucleus basalis magnocellularis.

The distribution of specifically-labeled neurotensin binding sites was examined in relation to that of cholinergic neurons in the rat nucleus basalis magnocellularis at both light and electron microscopic levels. Lightly prefixed forebrain slices were either labeled with [125I](Tyr3) neurotensin alone or processed for combined [125I]neurotensin radioautography and acetylcholinesterase histochemistry. In light microscopic radioautographs from 1-microns-thick sections taken from the surface of single-labeled slices, silver grains were found to be preferentially localized over perikarya and proximal processes of nucleus basalis cells. The label was distributed both throughout the cytoplasm and along the plasma membrane of magnocellular neurons all of which were found to be cholinesterase-positive in a double-labeled material. Probability circle analysis of silver grain distribution in electron microscopic radioautographs confirmed that the major fraction (80-89%) of specifically-labeled binding sites associated with cholinesterase-reactive cell bodies and dendrites was intraneuronal. These intraneuronal sites were mainly dispersed throughout the cytoplasm and are thus likely to represent receptors undergoing synthesis, transport and/or recycling. A proportion of the specific label was also localized over the nucleus, suggesting that neurotensin could modulate the expression of acetylcholine-related enzymes in the nucleus basalis. The remainder of the grains (11-20%) were classified as shared, i.e. overlied the plasma membrane of acetylcholinesterase-positive neuronal perikarya and dendrites. Extrapolation from light microscopic data, combined with the observation that shared grains were detected at several contact points along the plasma membrane of cells which also exhibited exclusive grains, made it possible to ascribe these membrane-associated receptors to the cholinergic neurons themselves rather than to abutting cellular profiles. Comparison of grain distribution with the frequency of occurrence of elements directly abutting the plasma membrane of neurotensin-labeled/cholinesterase-positive perikarya indicated that labeled cell surface receptors were more or less evenly distributed along the membrane as opposed to being concentrated opposite abutting axon terminals endowed or not with a visible junctional specialization. The low incidence of labeled binding sites found in close association with abutting axons makes it unlikely that only this sub-population of sites corresponds to functional receptors. On the contrary, the dispersion of labeled receptors seen here along the plasma membrane of cholinergic neurons suggests that neurotensin acts primarily in a paracrine mode to influence the magnocellular cholinergic system in the nucleus basalis.

Acetylcholinesterase↗

Cholinergic regulation of vasoactive intestinal peptide content and release in rat frontal cortex and hippocampus.

The purpose of this study was to determine whether the cholinergic system might have a regulatory role on vasoactive intestinal peptide (VIP) synthesis and release in the rat hippocampus and frontal cortex. Incubation of hippocampal or frontal cortical slices with the muscarinic agonist oxotremorine or antagonist atropine did not significantly alter VIP release. The nicotinic agonist methylcarbamylcholine (MCC) and the nicotinic antagonist dihydro-beta-erythroidine were also ineffective in altering VIP release. Chronic atropine (20 mg/kg, s.c., b.i.d., 10 days) and nicotine (0.59 mg/kg, s.c., b.i.d., 10 days) treatment significantly decreased the VIP content of the frontal cortex, by 42% and 26%, respectively. In contrast, neither treatment significantly altered the VIP content of the hippocampus. Both drug treatments decreased the amount of VIP released from tissue slices depolarized with veratridine in both cerebral cortex and hippocampus. Therefore, long-term treatment with atropine and nicotine results in changes in the synthesis and release of VIP in the cerebral cortex, whereas in the hippocampus the effect is limited to an alteration of VIP release. These results suggest that the acetylcholine regulates VIP neurotransmission in the rat frontal cortex and hippocampus by an action on muscarinic and nicotinic receptors.

Acetylcholine↗

Neurotensin regulation of endogenous acetylcholine release from rat cerebral cortex: effect of quinolinic acid lesions of the basal forebrain.

The effects of neurotensin (NT) on endogenous acetylcholine (ACh) release from basal forebrain, frontal cortex, and parietal cortex slices were tested. The results show that NT differentially regulates evoked ACh release from frontal and parietal cortex slices without altering either spontaneous or evoked ACh release from basal forebrain slices. In the frontal cortex, NT significantly inhibited evoked ACh release by a tetrodotoxin (TTX)-insensitive mechanism, suggesting an action directly on cholinergic terminals. In the parietal cortex, NT enhanced evoked ACh release by a TTX-sensitive mechanism, suggesting an action of NT on the cholinergic neuron or in close proximity to the cholinergic neuron. The effects of NT on ACh release were confined to evoked ACh release; that is, spontaneous ACh release was not affected. NT did not affect spontaneous or potassium-evoked ACh release from occipital cortex slices. The second set of experiments tested the effects of quinolinic acid (QUIN) lesions of the basal forebrain cell bodies on the NT-induced regulation of evoked ACh release in the cerebral cortex. QUIN lesions of basal forebrain cell bodies caused decreases in choline acetyltransferase activity (27 and 28%), spontaneous ACh release (14 and 21%), and evoked ACh release (38 and 44%) in frontal and parietal cortex, respectively. In addition, 11 days following QUIN lesions of basal forebrain cell bodies, the action of NT to regulate evoked ACh release in frontal cortex or parietal cortex was no longer observed. The results suggest that in the rat frontal and parietal cortex, NT differentially regulates the activity of cholinergic neurons by decreasing and increasing evoked ACh release, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylcholine↗

Cysteamine-induced enhancement of growth hormone-releasing factor (GRF) immunoreactivity in arcuate neurons: morphological evidence for putative somatostatin/GRF interactions within hypothalamus.

The episodic secretion of GH is regulated through the reciprocal release into the hypophyseal portal circulation of two hypothalamic peptides, SRIF and GRF. Recent physiological evidence suggests that, in addition to exerting their well documented opposite actions at the level of the anterior pituitary, SRIF and GRF may interact within the central nervous system to modulate GH secretion. The aim of the present study was to provide morphological evidence to support the concept that SRIF exerts an influence on the GRF-containing neuronal system within the hypothalamus. To accomplish this, we used the thiol agent cysteamine (CSH; 300 mg/kg sc) to manipulate endogenous hypothalamic SRIF and examined its effects on the immunoreactivity and mRNA content of GRF-containing arcuate (ARC) neurons, using immunohistochemistry and semi-quantitative in situ hybridization, respectively. Hypothalamic SRIF content and pulsatile GH secretion were also monitored. CSH treatment reduced hypothalamic immunoreactive SRIF concentrations (to 60% of vehicle-injected control values) and severely suppressed the spontaneous surges of GH release (mean GH peak amplitude: 24.2 +/- 2.7 vs. 168.0 +/- 27.8 ng/ml in H2O-injected controls; P less than 0.001). CSH treatment also resulted in a striking increase in the number and labeling density of GRF-immunoreactive cells detected in the ARC nucleus. Cell counts revealed an overall 126% increase, over controls, in the mean number of detectable GRF-positive ARC neurons. This increase (P less than 0.01) was apparent over all rostrocaudal levels of the ARC nucleus, but was most pronounced within the caudal tier. Administration of CSH also produced a massive qualitative increase in the intensity and extent of staining of GRF-positive terminal elements in the external zone of the median eminence. Conversely, CSH treatment resulted in a decline in GRF mRNA-associated signal in cells of the ARC nucleus, with this decline being most evident in the caudal third of the nucleus. The finding of an increase in GRF immunoreactivity in ARC neurons in association with a decrease in plasma GH, as well as with a decline in relative GRF mRNA content, suggests that CSH leads to an overall decrease in GRF release from the median eminence. These results provide morphological evidence to support the concept of a SRIF-mediated central influence on the GRF-containing ARC neuronal system. Such central regulation of GRF by SRIF may be an important mechanism in the physiological control of pulsatile GH secretion.

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Alterations in opioid parameters in the hypothalamus of rats with estradiol-induced polycystic ovarian disease.

The distribution and density of selectively labeled mu-, delta-, and kappa-opioid binding sites were examined by in vitro radioautography in the hypothalamus of normal, estradiol valerate (EV)-injected, and estradiol (E2)-implanted female rats. Hypothalamic beta-endorphin concentration was also examined by RIA in these three groups of animals. Quantitative analysis of film radioautographs demonstrated a selective increase in mu-opioid binding in the medial preoptic area of EV-treated, but not of E2-implanted rats. However, both these estrogenized groups exhibited a reduction in the density of delta-opioid binding in the suprachiasmatic nucleus. Statistically significant changes between either estrogenized groups were not observed for kappa-opioid binding. Results on the hypothalamic concentration of beta-endorphin indicated a marked reduction in EV-injected animals with respect to controls. In contrast, the E2-implanted animals exhibited beta-endorphin concentrations similar to controls. The present results confirm the increase in opioid receptor binding previously reported in the hypothalamus of EV-treated rats and further demonstrate that this increase is confined to the medial preoptic area and exclusively concerns mu-opioid receptors. The concomitant reduction in beta-endorphin levels observed in the same group of animals suggests that the observed increase in mu-opioid binding could reflect a chronic up-regulation of the receptor in response to compromised beta-endorphin input. Given the restriction of this effect to the site of origin of LHRH neurons and the demonstrated inhibitory role of opioids on LHRH release, it is tempting to postulate that such up-regulation could lead to the suppression of the plasma LH pattern that characterizes polycystic ovarian disease in the EV-treated rat.

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Electron microscopic localization of immunoreactive enkephalinase (EC 3.4.24.11) in the neostriatum of the rat.

The fine structural distribution of the enzyme-neutral endopeptidase EC 3.4.24.11 (enkephalinase) was examined by immunoradioautography (using an iodinated monoclonal antibody) and peroxidase immunocytochemistry (using the same probe in nonradioactive form) in the neostriatum of the rat. At the light microscopic level, both techniques revealed a heterogeneous distribution of immunoreactive enkephalinase in the caudoputamen, characterized by the presence of patches of intense immunolabeling prominent against a relatively strong immunoreactive matrix, a pattern reminiscent of mu opioid receptors radioautographically labeled in the same region. Pilot experiments indicated that fixation of the brain with a mixture of 4% paraformaldehyde, 0.05% glutaraldehyde, and 0.2% picric acid did not modify the distribution and only slightly reduced the intensity of striatal enkephalinase antigenicity, provided that the post-fixation period did not exceed 1 hr. In the neostriatum of animals fixed according to this protocol, enkephalinase immunoreactivity was found by electron microscopic immunoradioautography to be exclusively confined to neuronal and glial membrane interfaces. Immunoperoxidase cytochemistry confirmed the association of immunoreactive enkephalinase with the plasma membrane of neurons and, to a lesser extent, of astrocytes and oligodendrocytes. Both immunoradioautographic and immunoperoxidase techniques revealed a predominant association of the enzyme with neuronal perikarya and dendrites. The morphological features of the labeled perikarya, together with the presence of immunoreactive dendritic spines, suggested that some of these neurons corresponded to striatofugal medium spiny neurons. Immunoreactive enkephalinase was also detected at the level of myelinated and unmyelinated axons and axon terminals. These axons could potentially have originated from intrinsic striatal neurons or from the substantia nigra. Statistical analysis of silver grain distribution in electron microscopic immunoradioautographs indicated that immunoreactive enkephalinase was not preferentially concentrated at the level of specific membrane interfaces, but rather, was more or less uniformly distributed on the surface of neurons and/or glial cells. A similarly diffuse localization of the enzyme was apparent in peroxidase-reacted material, though the latter technique also revealed a microheterogeneity in the deposition of the reaction product along the labeled membranes. Finally, quantitative analysis of immunoradioautographs clearly indicated an absence of enkephalinase enrichment at the level of synaptic junctions. The similarity between the light and electron microscopic distribution of enkephalinase observed in the present study, and that previously reported for mu opioid receptors, lends support to the concept that this ectoenzyme may be involved in the inactivation of endogenous opioids in the mammalian neostriatum.

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Immunocytochemical evidence for direct connections between neurotensin-containing axons and dopaminergic neurons in the rat ventral midbrain tegmentum.

A light- and electron-microscopic double antigen localization technique was employed to study cellular relationships between neurotensin (NT)-containing axons and dopaminergic neurons in the substantia nigra and ventral tegmental area of the rat. Direct contacts were observed between NT-immunoreactive axon terminals and the dendrites and perikarya of neurons containing the dopamine biosynthetic enzyme, tyrosine hydroxylase (TH). NT-positive terminals were also found to contact unlabeled dendrites and neuronal somata. These results reveal an ultrastructural substrate through which endogenous NT might influence the activity of dopaminergic neurons in the ventral midbrain tegmentum.

Animals↗

Correspondence between high affinity 125I-neurotensin binding sites and dopaminergic neurons in the rat substantia nigra and ventral tegmental area: a combined radioautographic and immunohistochemical light microscopic study.

Several lines of anatomical, biochemical, and pharmacological evidence have suggested that specific high affinity neurotensin binding sites are associated with dopamine-containing neurons in the rat ventral tegmentum. In the present study we confirmed and quantified the extent of this association by combining monoiodinated neurotensin radioautography and tyrosine hydroxylase immunohistochemistry on adjacent 5-10 microns-thick midbrain sections. We found that 95-100% tyrosine hydroxylase-immunoreactive neurons detected in all subdivisions of the substantia nigra (pars compacta, pars lateralis, and pars reticulata) exhibited intense 125I-neurotensin labeling in adjacent light microscopic radioautographs. Tyrosine hydroxylase-positive dendrites radiating downward from compacta neurons were also heavily labeled throughout the pars reticulata. In the paranigral subdivision of the ventral tegmental area, silver grains were evenly distributed over neuropil and perikarya and therefore could not be readily attributed to any given tyrosine hydroxylase-positive element. In contrast, within the parabrachial pigmentous subdivision of the ventral tegmental area, 80-90% of the tyrosine hydroxylase-immunoreactive somata and proximal processes were clearly in register with 125I-neurotensin labeled cells. Finally, all tightly packed TH-positive neurons in the interfascicular nucleus showed intense 125I-neurotensin labeling. The vast majority of the neurotensin binding sites observed in the ventral midbrain tegmentum were of the high affinity, physiologically active receptor type since levocabastine, a selective blocker of the low affinity neurotensin binding component, had minimal effect on the binding density in any of the midbrain regions examined. The present results demonstrate an extensive overlap between specific, high affinity neurotensin binding sites and dopamine perikarya and dendrites in the rat ventral tegmentum, and thereby provide a direct anatomical substrate for observed neurotensin-dopamine interactions in the mesocorticolimbic and nigrostriatal projection systems.

Animals↗

Somatostatin receptors in brain and pituitary.

Somatostatin (SRIF) actions in the brain and pituitary are mediated by specific receptors. Using radioiodinated ligands it has been possible to characterize the kinetics of specific binding sites in the brain and pituitary, and to determine their cellular localization by autoradiography. At the pituitary level, the inhibition of growth hormone, prolactin and thyrotropin secretions induced by SRIF is mediated through a single binding site which is coupled to the inhibition of adenylate cyclase. In the brain, SRIF receptors are localized on neurons and glial cells and are also coupled to adenylate cyclase inhibition. Two sites are differentiated in the brain with an analogue of somatostatin, SMS 201995. In humans, SRIF-binding sites have been related to a number of pathologies. At the pituitary level, it has been shown that the number of binding sites was negatively correlated to growth hormone levels in acromegaly. Furthermore, SRIF-binding sites were undetectable in a patient which did not respond to SMS 201995 therapy. In the brain, meningiomas and gliomas are rich in SRIF binding sites. This suggests a possible role for SRIF on glia. In neurodegenerative diseases, cortical SRIF concentrations are decreased in Alzheimer's and Parkinson's disease associated with dementia while SRIF-binding sites are only affected in Alzheimer's disease. In conclusion, the physiological role of SRIF in the brain and pituitary can be evaluated by studying the receptors of the peptide. Such studies allow to question the implication of SRIF in endocrine and neuropathologies.

Animals↗

Electron microscopic localization of neurotensin binding sites in the midbrain tegmentum of the rat. I. Ventral tegmental area and the interfascicular nucleus.

The distribution of specifically labeled neurotensin (NT) binding sites was examined by light and electron microscopic radioautography in the ventral tegmental area (VTA) and nucleus interfascicularis of the rat following incubation of lightly prefixed midbrain slices with the monoiodinated ligand, 125I-(Tyr3)-NT. Film radioautograms of whole 125I-NT-incubated slices exhibited intense NT displaceable binding throughout the VTA and interfascicular nucleus. In light microscopic radioautographs from 1-microns-thick sections taken from the surface of the slices, the label was found to be present both inside and outside neuronal perikarya. Probability circle analysis of silver grain distribution in electron microscopic radioautographs confirmed that a significant proportion (greater than 20%) of the specifically labeled binding sites was intraneuronal. The frequent association of these sites with profiles of rough endoplasmic reticulum or Golgi apparatus suggested that they corresponded in part to receptors undergoing synthesis and/or glycosylation. The remainder was associated with neuronal and/or glial plasma membranes, as attested by comparing the distribution of grains overlying apposed cellular elements with the distribution of hypothetical grains originating from randomly distributed membrane bound radioactive sources. Although the resolution of the technique did not make it possible to ascribe labeled membrane-bound receptors to either one of the apposed plasma membranes, their frequent association with interfaces involving the plasmalemma of perikarya and dendrites, together with the occurrence of silver grain alignments along the membrane of certain somata and dendrites suggested that a proportion of them was associated with the perikarya and dendrites of a subpopulation of ventral tegmental neurons. Interestingly, these perikaryal and dendritic receptors were not exclusively present on, or even concentrated opposite, abutting axon terminals but instead were more or less evenly distributed along the plasma membrane. Only an exceedingly small proportion was found to be associated with synaptic junctions. Such a low incidence makes it unlikely that only the synapse-linked binding sites correspond to functional receptors. On the contrary, the dispersion of labeled receptors seen here along the plasma membrane of presumptive dopamine neurons suggests that NT acts mainly in a paracrine or parasynaptic fashion in the ventral midbrain tegmentum.

Animals↗