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L Descarries

Publications and source records attributed to L Descarries.

At least 91 records · Page 5Linked to original sources

Monoamine innervation of the oculomotor nucleus in the rat. A radioautographic study.

The serotonin and noradrenaline innervations of the rat oculomotor nucleus were examined by high resolution radioautography after in vivo labeling with tritiated 5-hydroxytryptamine and dopamine, respectively. Noradrenaline as well as serotonin endings (axonal varicosities) pervaded the entire nucleus, but the latter were at least six times more numerous (1.3 X 10(6) per mm3 of tissue) and were often found in the immediate vicinity of neuronal somata and proximal dendrites. The axon terminals of both types were of similar size and exhibited some large dense-cored vesicles in association with aggregated small and clear vesicles. The dense-cored vesicles were, however, more frequent and the content in clear vesicles more pleomorphic in serotonin than noradrenaline endings. In single thin sections, the proportion of noradrenaline and serotonin profiles exhibiting a synaptic junction was relatively small (15%). These were either symmetrical or asymmetrical when made on dendritic branches but invariably symmetrical on spines. In addition, a significant number of serotonin terminals were seen in close apposition or synaptic contact with neuronal perikarya and large dendrites, allowing for a direct, "proximal" action of serotonin. Moreover, many such terminals appeared to be coupled with unlabeled endings of another category, characterized by dispersed, uniformly round and clear synaptic vesicles, providing an alternate route for a proximal effect of serotonin in the oculomotor nucleus. In line with previous investigations on other motor nuclei, these data support the likelihood of a close involvement of both noradrenaline and serotonin in the control of motoneuronal activity.

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Quantification of the dopamine innervation in adult rat neostriatum.

Conditions leading to specific and integral visualization of dopamine axon terminals (varicosities) were tested in adult rat cerebral hemisphere slices incubated with [3H]dopamine and processed for high resolution radioautography. Specific visualization of the dopamine endings was achieved after incubation with 10(-6) M [3H]dopamine in the presence of a monoamine oxidase inhibitor (pargyline 10(-4) M), and of desipramine (5 X 10(-6) M), an inhibitor of catecholamine uptake by noradrenaline and serotonin neurons. [3H]Dopamine varicosity labeling was eliminated by the addition of 5 X 10(-5) M benztropine (an inhibitor of catecholamine uptake by catecholamine neurons), and was almost absent when dopamine nerve cell bodies of the midbrain had been previously destroyed with 6-hydroxydopamine. In dopamine-denervated neostriatum incubated without desipramine, a second set of labeled terminals was also visible. These were identified as serotoninergic, since their labeling was suppressed by citalopram, an inhibitor of monoamine uptake highly specific for serotonin neurons. There was no desipramine-sensitive but citalopram-resistant varicosity labeling suggestive of neostriatal noradrenaline innervation. In normal striatum, incubation at 35 degrees C always resulted in a labeling of dopamine varicosities restricted to a narrow band which followed the contours and cut surface of this anatomical region. This unusual distribution was the result of an uptake barrier generated by the tightly packed dopamine varicosities. Indeed, the striatal dopamine varicosity labeling was more widespread after partial 6-hydroxydopamine denervation or in normal tissue incubated either with a higher [3H]dopamine concentration (5 X 10(-6) M), in the presence of relatively low benztropine concentrations (10(-5) M), or at lower temperature (15 degrees C). Material incubated at 15 degrees C for 90 min was suitable for purposes of quantification: labeled varicosities were then visualized throughout the striatum and across the full thickness of the slices; moreover, the number of labeled varicosities plotted against radioautographic exposure time increased in parallel and reached a plateau at the same time in neostriatal sectors with widely different innervation densities. At a rostral transverse level across neostriatum, the dorsolateral quadrant showed hyperdense "patches" of labeled terminals distinguishable from an already dense surrounding "matrix", whereas, ventromedially, the dopamine innervation appeared more uniform and somewhat less dense.(ABSTRACT TRUNCATED AT 400 WORDS)

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Distribution of [3H]noradrenaline and [3H]serotonin in photophores of porichthys notatus. An electron-microscopic radioautographic analysis.

Photophores of Porichthys notatus were examined by electron-microscopic radioautography following incubation in tritiated noradrenaline ( [3H]NA) or serotonin ( [3H]5-HT). Nerve varicosities surrounding the photocytes were found to accumulate [3H]NA but not [3H]5-HT, providing compelling evidence for the catecholaminergic nature of the monoaminergic innervation of photophores. The photocytes themselves appeared selectively labelled with both tracers, but the intensity of labelling after [3H]5-HT incubation was considerably greater than after [3H]NA. Stereological sampling of organelle content in photocytes showed ultrastructural differences between [3H]NA- and [3H]5-HT-labelled cells, probably related to light emission induced by NA. The main changes noted after incubation with [3H]NA were mitochondrial swelling and disorganization, increased coalescence of photocytic vesicles and extrusion of vesicular material into the extracellular matrix. With respect to the subcellular localization of [3H]NA and [3H]5-HT within the photocytes, statistical analysis of the distribution of silver grains disclosed a preferential affinity of both labels for appositional zones between mitochondria and coalescent vesicles. Moreover, in the case of 5-HT, selective affinity was also exhibited by sites comprising vesicular membrane and adjacent cytoplasm, suggesting binding of this biogenic amine to the entire membrane of photocytic vesicles.

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Uptake and retention of [3H]adrenaline by central monoaminergic neurons: a light- and electron-microscope radioautographic study after intraventricular administration in the rat.

Paraventricular and paracisternal regions of adult rat central nervous system were investigated by light- and electron-microscope radioautography after intraventricular administration of tritiated adrenaline. In tissue primarily fixed by glutaraldehyde perfusion and post-fixed by immersion in osmium tetroxide, there were no aggregates of silver grains indicative of intraneuronal accumulation of the tracer, except over perivascular nerve terminals at the base of the brain. In contrast, when both fixation and postfixation were carried out by rapid vascular perfusion, preferentially labeled nerve cell bodies and axonal varicosities (i.e. terminals) were detected in various anatomical areas known to contain dopaminergic and/or noradrenergic neurons. Serotoninergic axonal varicosities in the supraependymal plexus and subcommissural organ, as well as a small group of nerve cell bodies of undetermined chemical identity in the n. paraventricularis thalami were also found to be labeled. Addition of a ten-fold higher concentration of non-radioactive serotonin to the solution of [3H]adrenaline suppressed the reactivity in the subcommissural organ and the supraependymal plexus but had no such effect elsewhere in brain. Lesioning of the nigrostriatal dopaminergic system with 6-hydroxydopamine prior to [3H]adrenaline injection eradicated axon terminal labeling in the ipsilateral neostriatum. Electron-microscopic examination of [3H]adrenaline-labeled varicosities in the neostriatum, lateral septum, arcuate nucleus and median eminence extended earlier observations on the ultrastructure of the catecholaminergic innervation of these regions. It was concluded that both dopaminergic and noradrenergic neurons as well as certain serotonin-containing axon terminals can take up and retain [3H]adrenaline, although they probably have lesser affinity for this amine than for their own transmitter. Due to the fact that presumptive adrenergic neurons are intermingled with dopaminergic and noradrenergic elements, further work will be needed to determine to which extent they also contributed to [3H]adrenaline uptake in the present experimental conditions.

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The serotonin neurons in nucleus raphe dorsalis of adult rat: a light and electron microscope radioautographic study.

The serotoninergic nerve cell body population of nucleus raphe dorsalis (RD) was identified by radioautography following cerebroventricular instillation of tritiated serotonin ([3H]5-HT) in adult rats pretreated with a monoamine oxidase inhibitor. Series of histological sections taken throughout the midbrain and upper pons exhibited a similar distribution and number of labeled nerve cell bodies in RD after prolonged administration of either 10-5 or 10-4M [3H]5-HT or 10-4M [3H]5-HT and 10-3M nonradioactive noradrenaline. This allowed systematic mapping and quantification of serotoninergic nerve cell bodies at various levels of the RD. Their extrapolated total number averaged 11,500. Twice as many unreactive (nonserotoninergic) neurons were present within the same region. In electron microscope radioautographs, the labeled cells were usually larger (17.9 micrometer mean diameter) than their unlabeled congeners (13.1 micrometer), but stereological sampling of their perikarial organelle content failed to reveal any difference in cytoplasmic composition. Few [3H]5-HT-labeled axonal varicosities were observed in RD and none were found in close apposition or in synaptic junction with labeled nerve cell bodies, dendrites, or unreactive perikarya. A detailed statistical analysis of silver grain distribution in both labeled and "unlabeled" nerve cell bodies, indicated that in the former, but not in the latter, dense bodies had a relatively high affinity for [3H]5-HT. Mitochondria and the cytoplasmic membrane were the only other organelles to show higher labeling indices in labeled than in unlabeled cells. Other sites of [3H]5-HT localization could be ascribed to artefactitious cross-linkage of the tracer by the fixative, since they had the same relative affinity in the two cell populations. These results provide new insights into the morphology and cytofunctional properties of the 5-HT neurons of rat RD.

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Altered neuronal responsiveness to biogenic amines in rat cerebral cortex after serotonin denervation or depletion.

To further investigate monoaminergic mechanisms in cerebral cortex, responsiveness of cortical neurons to microiontophoretic applications of serotonin (5-HT), dopamine (DA) or noradrenaline (NA) was examined in the frontoparietal region of control, 5,7-dihydroxytryptamine (5,7-DHT)- and p-chlorophenylalanine (PCPA)-treated rats anesthetized with urethane. As a rule, 100 nA applications of either one of these biogenic amines induced marked slowing or total interruption of 'spontaneous' firing overlasting the 30 s period of ejection. Given the large amounts of monoamines ejected, it could be inferred that such microiontophoretic applications produced a maximal activation of receptors. In control rats, the responses to 5-HT, DA and NA were of approximately equal duration (approximately equal to 5 min). Two to 4 weeks after denervation with 5,7-DHT, most neurons (75%) exhibited greatly prolonged responses to 5-HT (approximately equal to 14 min), and marked depressions of firing could be induced by small ejection currents (approximately equal to 2 nA) having little or no effect in the controls. In addition, 85% of the units supersensitive to 5-HT showed considerably shortened responses to DA and NA (approximately equal to 1 min). After 2-14 days of depletion with PCPA, there was no change in the responsiveness to 5-HT in spite of a 91% lowering of cortical 5-HT content equivalent to that measured after denervation. Nevertheless, responsiveness to DA and NA was again diminished in a majority (80%) of the units tested. In control or PCPA-treated rats, acute administration of the 5-HT re-uptake blocker fluoxetine increased the duration of depressions induced by 100 nA applications of 5-HT but did not enhance responsiveness to low ejection currents. This suggested that, after 5-HT denervation, the suppression of re-uptake was mainly responsible for the prolongation of 5-HT responses ('presynaptic' component of supersensitivity), whereas a modification of 5-HT receptors accounted for the greater efficacy of small doses of 5-HT ('postsynaptic' component). Responsiveness to the microiontophoretic application of phenylephrine (PHE), a noradrenergic a-agonist, was comparable with that to NA in PCPA- and 5,7-DHT-treated as well as in control rats. Therefore, the hyposensitivity to DA and NA appeared indicative of a desensitization of catecholamine receptors caused by the absence of 5-HT. Such a desensitization may be viewed as an adaptive change resulting from an increased release of endogenous DA and NA. This interpretation would in turn imply that, normally, 5-HT regulates catecholamine release in the neocortex.

Afferent Pathways↗

Catecholamines and 5-hydroxytryptamine in photophores of Porichthys notatus. Radioenzymatic detection and radioautographic localization.

Radioenzymatic assays and light microscope radioautographic studies performed on photophores of Porichthys notatus demonstrated (1) significant amounts of catecholamines (dopamine, noradrenaline, adrenaline) and 5-hydroxytryptamine (serotonin) in these organs; (2) selective uptake and storage of [3H]noradrenaline ([3H]NA) by axon terminals innervating the photocytes, and (3) strong accumulation of [3H]5-hydroxytryptamine ([3H]5-HT) within the photocytes. Uptake and storage of [3H]NA in the nerve fibers were seemingly unaffected by the addition of ten-fold molar concentrations of unlabelled serotonin. Accumulation of [3H]5-HT by the photocytes was dose-dependent and diminished markedly in the presence of ten-fold molar concentrations of non-radioactive noradrenaline. Neither neuronal uptake of [3H]5-HT or [3H]A, nor photocytic accumulation of [3H]A were detectable under the conditions of the present experiments. This information should provide a framework for further investigations of the regulation of photophore luminescence by the biogenic amines.

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Immunocytochemical detection of peptides in osmicated and plastic-embedded tissue. An electron microscopic study.

In an attempt to improve the ultrastructural preservation of tissue immunostained with the postembedding staining technique, we verified the effect of postfixation with OsO4 on the antigenicity of several pituitary hormones and neuropeptides. For this purpose, the rat pituitary and central nervous system (CNS) were perfused with 2.5% glutaraldehyde and postfixed in two different ways: a) postfixation by immersion of small fragments in a mixture of 1% OsO4 and 1% potassium ferrocyanide, and b) postfixation with perfusion of 500 ml of 0.5% OsO4. The results obtained were similar with the two types of postfixation. In the pituitary gland, all the hormones could be very easily detected, although the staining was less intense for the glycoprotidic hormones. In the CNS the following neuropeptides: somatostatin, luteinizing hormone-releasing hormone (LHRH), Leu-enkephalin and substance P could be immunostained. These results indicate that a variety of polypeptidic antigens, including small neuropeptides, can survive postfixation with OsO4. In both the pituitary and CNS, the ultrastructural preservation was very good with a high contrast of membranes, thus permitting a clear identification of positive organelles. Moreover, it appears that, since the synaptic junctions are well-preserved after postfixation with OsO4, postembedding staining can now be utilized to identify and classify the different categories of endings containing neuropeptides.

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Responsiveness of cortical neurons to serotonin after 5,7-DHT denervation or PCPA depletion.

1. In adult rats under urethane anesthesia, a vast majority of spontaneously active neurons in the frontoparietal cortex undergo a prolonged depression of their firing rate upon microiontophoretic application of 5-HT. 2. In 5,7-DHT-deafferented cortex, this effect is of a longer duration (mean 14 min) than in controls (mean 5 min). Moreover, small ejection currents of 5-HT are sufficient to induce a prolonged depression of the firing rate. 3. In PCPA-pretreated rats, there are no changes in the responsiveness to 5-HT. 4. In control and PCPA-pretreated rats, blocking of the 5-HT reuptake with fluoxetine increases the duration of responses to 5-HT (mean 15 min), whereas small ejection currents remain without effect. 5. These data indicate that, in the cerebral cortex, denervation supersensitivity to 5-HT results primarily from the removal of 5-HT afferents, and not from depletion of their 5-HT content. The enhanced responsiveness to microiontophoresed 5-HT appears to be due to a suppression of reuptake mechanisms, mainly responsible for the prolongation of 5-HT effects, and to a modification of receptors on target cells, which accounts for their greater sensitivity to 5-HT.

5,7-Dihydroxytryptamine↗

[The dopaminergic innervation of neostriatum: a new possibility of radioautographic investigation (author's transl)].

Precise knowledge of the ultrastructural features and interneuronal relationships of dopaminergic (DA) axon terminals or varicosities in neostriatum is still lacking. This ignorance is due to current limitations of the methods applicable to their specific visualization at electron microscopic level. High resolution radioautography, in particular, has not yet permitted a clearcut identification of the DA nerve endings which take up and store exogenous catecholamines in vivo, due to an apparent mobilization of tracer during standard histological preparative procedures for light and electron microscopy (Fig. 1 A). In this context, histological processing of the central nervous system by vascular perfusion, tested in adult rats subjected to prolonged lateroventricular instillation of [3H]DA, led to the following results and conclusions: 1 Axonal varicosities having accumulated [3H]DA in vivo may be detected in great number in the ipsilateral paraventricular neostriatum (Figs. 1 B and 1 C). 2 The specificity of this radioautographic labelling is evidenced by: (a) its disappearance or persistance, depending on the addition of a high concentration of non-radioactive noradrenaline or serotonin to the [3H]DA solution (Fig. 1 C); (b) the absence of labelled axonal varicosities in the supraependymal region (Figs. 2 E and 2 F) and suprachiasmatic nucleus (Fig. 1 E to be compared with 1 F) after administration of [3H]DA alone; (c) the absence of any localized accumulation of [3H]DA in neostriatum following prior destruction of the nigro-striatal DA system by 6-hydroxydopamine (Fig. 1 D). 3 It seems that carrying out the double fixation with glutaraldehyde and osmium by vascular perfusion is the prerequisite for retaining in situ the [3H]DA accumulated in vivo by neostriatal nerve endings. 4 Preliminary ultrastructural examination shows that the DA axonal varicosities of paraventricular neostriatum are of small caliber (mean diameter: 0.5 micron) and mostly contain clear synaptic vesicles, occasionally associated with a few larger dense-core vesicles (figs. 2 A-D). Several of these nerve endings establish axo-dendritic synaptic junctions (Fig. 2 C) and a few, perhaps, axo-somatic contacts (Fig. 2 D).

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The fine structure of central serotonin neurons.

1. Most of the current knowledge concerning the fine structure of serotonin (5-HT) neurons in mammalian CNS is derived from radioautographic studies following uptake of tritiated 5-HT in vivo. 2. (3H)5-HT-labelled perikaria and proximal dendrites have thus far been visualized ultrastructurally in nuclei raphe dorsalis, paragigantocellularis lateralis and dorsomedialis hypothalami. In none of these regions were they found to possess cytological features which would constitute a hallmark of their serotoninergic nature. 3. Most central 5-HT neurons give rise to thin and unmyelinated, albeit long and ramified axons. These innervate the entire neuraxis, issuing billions of varicose terminals along their paths. Such varicosities usually consist of small axonal enlargements (0.5-1 micrometers in diameter) containing clustered vesicular organelles. These organelles include small (15-50 nm), pleomorphic agranular vesicles and larger (50-130 nm), round or flattened dense-cored vesicles. 4. A salient feature of central 5-HT varicosities is that they often lack the junctional complexes classically considered as the morphological substrate for chemical transmission in CNS. The occurrence of synaptic specializations varies from one brain region to another. When present, synaptic contacts are generally found on dendritic branches or spines of nonserotoninergic neurons, but axo-somatic or axo-axonic synapses, as well as contacts with 5-HT elements, have also been reported. The frequency with which 5-HT varicosities establish junctional relationships may be more dependent on the territory of innervation than characteristic of any subset of 5-HT fibers or nerve cell bodies. 5. It is currently assumed that both non-junctional and junctional 5-HT varicosities can release their endogenous amine upon axonal depolarization. This mode of action, which evokes that of a local neurohormone, might subtend the modulatory effects of 5-HT in the CNS.

Afferent Pathways↗

Description of an indolaminergic cell component in the cat locus coeruleus: a fluorescence histochemical and radioautographic study.

Using Falck-Hillarp fluorescence histochemical and radioautographic techniques, it has been found that, in addition to the well-known catecholaminergic cells, the locus coeruleus (LC) of the cat contains a sizeable component of indolaminergic neurons. Indolaminergic cell bodies occur in all subdivisions of the LC complex. They are most numerous in the LC proper and subcoeruleus area, but are also present in the medial and lateral parabrachial, and Kölliker-Fuse nuclei. In all, the indolaminergic cells are estimated to make up 7-10% of the monoaminergic neuronal population of the LC complex. With the exception of the Kölliker-Fuse nucleus, where somewhat larger cells occur, the indolaminergic cell bodies in different parts of the LC complex share a common fluorescence histochemical appearance. They display round to fusiform shapes and measure 30 x 18 micron on the average, which makes them cytoarchitectonically similar to the small type of noradrenergic cells in the LC. The formaldehyde-induced fluorescence of the indolaminergic cells in the LC complex was analyzed microspectrofluorometrically and the recorded excitation and emission spectra (maxima at 370 and 530 nm, respectively) were found to be identical with those recorded from midline raphe neurons. No evidence of noradrenaline content was found in the indolaminergic cells of the LC. Radioautographic experiments after intratissular injections of tritiated serotonin showed that the indolaminergic cells of the LC complex possess uptake mechanisms for serotonin. Taken together these results provide strong evidence for serotonin being the transmitter of the indolaminergic neurons discovered in the LC of the cat.

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Modulatory role for biogenic amines in the cerebral cortex. Microiontophoretic studies.

In order to investigate the mode of action of biogenic amines in rat cerebral cortex, the unitary activity of spontaneously firing neurons and their excitatory response to acetylcholine (ACh) were examined using microiontophoretic administration of dopamine (DA), noradrenaline (NA) and serotonin (5-HT). The predominant effect of these biogenic amines on the spontaneous activity was a profound and prolonged inhibition of firing (2-4 min), which attained its maximum within 15-120 sec. This response was generally more abrupt in onset and of greater magnitude with NA and 5-HT than with DA. Most units inhibited by DA, NA and 5-HT also showed marked depression of their excitatory response to ACh when pretreated with these biogenic amines. With repetitive administration of ACh, it could be shown that the total duration of inhibition of ACh responses by DA and NA was not as prolonged as the inhibition of the spontaneous firing of the same cells. With 5-HT, the initial ACh responses of many neurons could be completely blocked, and this inhibitory effect lasted as long as the inhibition of spontaneous firing. In view of the anatomical data demonstrating a relative sparsity of monoamine nerve terminals in cerebral cortex, the strong inhibition induced by DA, NA or 5-HT may have reflected slow inactivation of the biogenic amines. However, it could also be indicative of underlying mechanisms of action dependent on metabolic changes. Indeed, the interaction between biogenic amines and ACh might imply a balance between the intracellular pools of cAMP and cGMP is directly or indirectly influenced by the biogenic amines and ACh, respectively. This hypothesis would not exclude other modes of local interaction between DA, NA, 5-HT and ACh, and appears compatible with the modulatory role of biogenic amines in cerebral cortex.

Acetylcholine↗