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M Geffard

Publications and source records attributed to M Geffard.

At least 145 records · Page 8Linked to original sources

Immunocytochemical demonstration of octopamine-immunoreactive cells in the nervous system of Locusta migratoria and Schistocerca gregaria.

The distribution of octopamine in the metathoracic ganglion, brain and corpus cardiacum of Locusta migratoria and Schistocerca gregaria was investigated by means of immunocytochemistry with an antiserum against octopamine. The dorsal unpaired median (DUM) cells of the metathoracic ganglion were found to be strongly octopamine-immunoreactive. In the rostroventral part of the protocerebrum a group of seven immunopositive cells was demonstrated. Stained nerve fibres of these cells run into three directions: circumoesophageal connectives, midbrain, and optic lobes. As far as the protocerebrum is concerned, immunoreactive fibres were found in the central body, the protocerebral bridge, and in other neuropile areas. In the optic lobe a dense plexus of immunopositive fibres was found in the lobula and in the medulla. In the brain one other immunopositive cell was demonstrated, situated at the lateral border of the tritocerebrum. Octopamine could not be shown to occur either in the globuli cells of the mushroom bodies or in the dorsolateral part of the protocerebrum, where the perikarya of the secretomotor neurones are located that innervate the glandular cells of the corpus cardiacum. In the nervi corporis cardiaci II, which contain the axons of the neurones that extend into the glandular part of the corpus cardiacum, and in the corpus cardiacum proper no specific octopamine immunoreactivity could be found.

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The use of Quetol 651 for the post-embedding immunohistochemical demonstration of gamma-aminobutyric acid on semithin sections.

Quetol 651 was used as an embedding medium for the demonstration of gamma aminobutyric acid (GABA) in semithin sections by the peroxidase-anti-peroxidase method. In order to demonstrate the immunoreactivity, the embedding medium was partially dissolved using absolute ethanol containing 0.8-1 M NaOH or KOH for 5-7 min. The experimental procedure was elaborated by testing the GABAergic sites in the endings surrounding the small neurones of the anterior exterolateral nucleus of a mormyrid fish and in the pyramidal cells of the electrosensory lateral line lobe of gymnotoid fish by applying anti-GAD (glutamic acid decarboxylase) antiserum. To test the general validity of the use of Quetol 651, GABAergic sites were also identified in the central nervous system of an insect, the honey bee, with anti-GABA and anti-GAD antisera. The intensity of labelling revealed by immunoperoxidase applied to Quetol 651-embedded semithin sections, demonstrated high precision and gave good resolution for light microscopical observations.

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Immunoelectron microscopy identifies several types of GABA-containing efferent synapses in the guinea-pig organ of Corti.

Using an immunoperoxidase technique, we have localized by light and electron microscopy GABA-immunostained fibers within a component of the efferent innervation of the organ of Corti. At the light microscopic level, GABA-immunostained fibers were observed within the inner spiral bundle (below the inner hair cells) and the tunnel spiral bundle. The immunostaining was clearly more intense in the upper turns than in the basal turns. Mostly in the upper turns, GABA-immunostained fibers were seen crossing the tunnel of Corti to reach the outer hair cells where they formed large immunostained patches at the base of the cells. Unevenly distributed throughout these upper turns, immunostained fibers were seen climbing along the outer hair cells and traveling near the non-sensorineural Hensen's cells. The electron microscopic observations of GABA-immunostained fibers in the upper turns allowed us to identify within the inner spiral bundle vesiculated varicosities synapsing with radial dendrites connected to the inner hair cells. In the outer hair cell area, the GABA-immunostained fibers made several kinds of synaptic contacts. They included a minor population of the large axosomatic synapses with the basal pole of the outer hair cells and many axodendritic synapses with the spiral dendrites connected to these cells. Occasionally, the GABA-immunostained climbing fibers also synapsed with the outer hair cells at a supranuclear level. These result confirm previous light microscopic data dealing with the projection of the GABA-immunostained fibers along the cochlear partition. Moreover, they extend them in characterizing several kinds of GABA-immunostained synapses. These latter findings agree with previous neurochemical electrophysiological data which suggests an efferent neurotransmitter role for GABA. Nevertheless, such an existence of an efferent innervation predominantly projecting to the upper turns of the cochlea adds another criterion distinguishing the "apical" from the "basal" cochlea.

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Glutamic acid decarboxylase- and gamma-aminobutyric acid-like immunoreactivities in corticotropin-releasing factor-containing parvocellular neurons of the hypothalamic paraventricular nucleus.

The indirect immunofluorescence technique was used to study the relation between corticotropin-releasing factor (CRF) and GABAergic neurons in the rat hypothalamic paraventricular nucleus (PVN). In colchicine-pretreated animals, glutamic acid decarboxylase (GAD)- and GABA-immunoreactive (IR) neurons were observed within the medial part of the parvocellular division of the PVN as well as surrounding the nucleus itself. In general, the GAD antiserum, as compared to the GABA antiserum, revealed stronger IR cells and a higher number of cells in the PVN. CRF-IR cells were observed throughout the whole rostrocaudal extension of the PVN, but predominantly in its dorsal medial parvocellular part. Adjacent sections stained, respectively, with antisera against CRF, GAD or GABA, revealed overlapping distributional patterns within the parvocellular part of the PVN. Employing a direct double-staining technique with sheep GAD and rabbit CRF antisera, it was demonstrated that both GAD and CRF immunoreactivities occurred in the same neurons, particularly in the dorsomedial aspect of the parvocellular PVN. GAD-positive neurons located in the ventromedial parvocellular PVN, periventricular nucleus and surrounding the PVN lacked CRF immunoreactivity. In addition, with an elution-restaining technique it was possible to demonstrate that also GABA immunoreactivity was present in some CRF neurons in the dorsomedial parvocellular PVN. Intravenous injection of the retrograde tracer Fast Blue resulted in labelling of neurons in the periventricular area and in both the parvocellular and magnocellular division of the PVN. After processing the sections for GAD/CRF immunohistochemistry, it was possible to demonstrate the presence of retrogradely labelled GAD/CRF cells, pointing at a PVN median eminence projection for these neurons. The present findings demonstrate that a subpopulation of the CRF-containing neurons in the parvocellular division of the PVN in addition are GABAergic, thereby indicating a possible corelease of these compounds, presumably in the median eminence. Possible interactions of GABA within the hypothalamo-pituitary-adrenal axis are discussed.

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Immunocytochemical localisation of GABA in endocrine cells of the rat entero-pancreatic system.

The occurrence of GABA-containing cells in the rat entero-pancreatic system was investigated by using anti-GABA-glutaraldehyde antibodies at the light and electron microscope level. In the pancreas, the B cells showed intense immunoreactivity, contrary to non-B and exocrine cells. Moreover, post-embedding immunogold staining was localised mostly in mitochondria, close to rough endoplasmic reticulum and in the nucleus. The insulin granules appeared nonsignificantly stained, which suggests the lack of cosecretion of GABA together with insulin. In the duodenum, GABA immunoreactivity was detected in certain endocrine cell types, suggesting a possible interaction with this amino acid. The well established GABAergic innervation in the enteric system was also confirmed by immunolabelling.

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First characterization of 5-hydroxytryptophan in rat brain by using specific antibodies.

DL-5-Hydroxytryptophan (5-HTP) was conjugated to bovine serum albumin and human serum albumin with glutaraldehyde (G). These conjugates made it possible to raise specific antisera in two rabbits. Their specificity and affinity were evaluated using an enzyme-linked immunosorbent assay and immunocytochemistry. For two antisera obtained, the most immunoreactive antigen was 5-HTP-G-protein, indicating that the same immune response was developed. The other conjugated indoleamines (5-methoxytryptophan-G-protein, tryptophan-G-protein) were poorly recognized or not at all (5-methoxytryptamine-G-protein, serotonin-G-protein, tryptamine-G-protein). These 5-HTP antisera enabled us to specifically visualize the precursor of serotonin in the raphe nuclei of G-fixed rat brains.

5-Hydroxytryptophan↗

5-Hydroxytryptophan (5-HTP)-immunoreactive neurons in the rat brain tissue.

We demonstrated the presence of 5-hydroxytryptophan (5-HTP), the immediate precursor of serotonin (5-HT), in the rat brain tissue using a glutaraldehyde-coupled immunohistochemical technique. The immunoreactivity of 5-HTP was intensified in the colchicine-pretreated rat. The distribution of labelled cells was the same as for 5-HT-immunoreactive cells, but they were fewer in number.

5-Hydroxytryptophan↗

[Anti-acetylcholine and anti-receptor antibodies in myasthenia].

The presence of anti-acetylcholine antibodies has been demonstrated in 24 patients with myasthenia gravis. Their titer was moderately but significantly higher than in controls. There was no significant correlation between the titer of these anti-acetylcholine antibodies and the clinical severity of the disease or the titer of anti-receptor antibodies in the same patients. Several populations of antibodies are present in myasthenia gravis according to an idiotype-anti-idiotype process, reflecting the complexity of immune reactions.

Acetylcholine↗

Natural seric anti-fatty acid antibodies in multiple sclerosis.

Naturally occurring anti-oleic acid conjugate antibodies were detected in human sera using an adapted direct immunoenzymatic assay. They were present to a higher level in the sera of patients with multiple sclerosis in acute relapse compared to patients with other neurological diseases or healthy subjects and even patients with multiple sclerosis in progressive form.

Adult↗

Dopamine-GABA interactions in the nucleus accumbens and lateral septum of the rat.

The relationships between dopaminergic afferents and GABAergic neurones were studied at the electron microscopic level in the rat lateral septum and nucleus accumbens by coupling 6-hydroxydopamine degeneration and gamma-aminobutyric acid (GABA) immunocytochemistry. Degenerating fibres were observed in the two regions making synaptic contact with GABA-immunoreactive and non-labelled cell bodies and dendrites. It is concluded that dopaminergic afferents to the septum and the nucleus accumbens contact, among others, a population of GABAergic cells. A similar route of regulation of the basalo-cortical and septo-hippocampal cholinergic pathways by dopaminergic afferents is proposed.

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Immunocytochemical localization of acetylcholine receptors in locust brain using auto-anti-idiotypic acetylcholine antibodies.

Auto-anti-idiotypic antibodies have been detected in antisera of rabbits immunized with an acetylcholine (ACh) conjugate. These antibodies were found to bind to ACh receptor (ACh-R) purified from different species membranes. They competed with the ACh-R antagonist alpha-bungarotoxin and some agonists such as ACh conjugate and ACh itself. They did not recognize acetylcholinesterase. Their characterization 'in vitro' suggested their employment as an immunohistological marker for ACh-R. In the locust brain, specific immunoreactivity was found in neuropils of the protocerebrum, the optic lobes, the deutocerebrum and the tritocerebrum.

Acetylcholine↗

Comparative study of dopamine- and noradrenaline-immunoreactive terminals in the paraventricular and supraoptic nuclei of the rat.

The distribution of dopaminergic and noradrenergic terminal fields of the paraventricular (PVN) and supraoptic (SON) nuclei of the rat was investigated at the optic and electron microscopical level using antibodies directed against dopamine (DA) and noradrenaline (NA). The DA innervation was uniform among these nuclei, although more important in the PVN than in the SON. NA fibers were preferentially distributed in the parvocellular parts of the PVN and in areas of the magnocellular nuclei where vasopressinergic neurons were mainly located. Both DA and NA terminals synaptically contacted magnocellular neurons on their cell body or dendrites. This study thus provides morphological evidence for a double and independent catecholaminergic control, by DA and NA, on neuroendocrine mechanisms at the hypothalamic level.

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Morphological evidence for a direct neuroendocrine GABAergic control of the anterior pituitary in teleosts.

The anterior pituitary of teleosts is unique among vertebrates in receiving a direct innervation which represents the morphological support of the neuroendocrine control of pars distalis functions. The participation of GABAergic fibers in this innervation was studied by means of immunocytochemistry at the light and electron microscopic levels, using antibodies against GABA. Immunoreactive fibers, characterized by the presence of small clear and dense cored vesicles, were detected in all parts of the gland. Immunopositive terminals were found in close, sometimes synaptic-like, contact with most glandular cell types in the anterior lobe. The data strongly suggest that in teleosts, as in mammals, GABA is involved in the neuroendocrine control of anterior pituitary functions.

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Convergence of electrotonic club endings, GABA- and serotoninergic terminals on second order neurons of the electrosensory pathway in mormyrid fish, Gnathonemus petersii and Brienomyrus niger (Teleostei).

Previous electrophysiological data indicate that the afferent electrosensory impulses conveyed towards the mesencephalon are blocked in the rhombencephalic electrosensory lateral line nucleus (nELL) by the concomitantly occurring EOD (electric organ discharge) command-associated (corollary) discharge. Electron-microscopic observations and anterograde labeling with horseradish peroxidase show that the primary electrosensory fibers terminate with club endings on the adendritic soma of the nELL cells and form gap junctions with the postsynaptic membrane. The remaining part of the soma and the initial segment membrane of nELL cells are covered with a large number of boutons showing chemical synaptic profiles. The GABA-ergic (gamma-aminobutyric-acid) nature of the majority of the boutons is revealed immunocytochemically by anti-GABA and anti-glutamic acid decarboxylase (anti-GAD) antisera, as seen in the light microscope. Electron-microscopic examination confirms the GABAergic nature of most of the bouton-like terminals, whereas club endings show negative immunoreactivity. In addition, serotonin-immunoreactive fibers and boutons are found in the same nucleus, between and next to the nELL cells. It is suggested that the GABAergic endings are the morphological basis for the inhibition that occurs in the nELL and that is mediated by the corollary discharge.

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Immunocytochemical study of the GABAergic innervation of the mouse pituitary by use of antibodies against gamma-aminobutyric acid (GABA).

The GABAergic innervation of the mouse pituitary, including the median eminence, was studied at light-microscopic and ultrastructural levels by use of a pre-embedding immunocytochemical technique with antibodies directed against GABA. In the median eminence, a high density of GABA-immunoreactive fibers was found in the external layer where the GABAergic varicosities were frequently observed surrounding the blood vessels of the primary capillary plexus. In the internal and subependymal layers, only few fibers were immunoreactive. The intense labeling of the external layer was observed in the entire rostro-caudal extent of the median eminence. In the pituitary proper, a dense network of GABA-immunoreactive fibers was revealed throughout the neural and intermediate lobes, entering via the hypophyseal stalk. The anterior and tuberal lobes were devoid of any immunoreactivity. The GABA-immunoreactive terminals were characterized in the median eminence, and in the intermediate and posterior lobes at the electron-microscopic level. They contained small clear vesicles, occasionally associated with dense-core vesicles or neurosecretory granules. In the intermediate lobe they were seen to be in contact with the glandular cells. In the posterior lobe and in the median eminence, GABA-immunoreactive terminals were frequently located in the vicinity of blood vessels. These results further support the concept of a role of GABA in the regulation of hypophyseal functions, via the portal blood for the anterior lobe, directly on the cells in the intermediate lobe, and via axo-axonic mechanisms in the median eminence and posterior lobe.

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Simultaneous detection of indoleamines and dopamine in rat dorsal raphe nuclei using specific antibodies.

Using a monoclonal antibody against dopamine and a rabbit antiserum against serotonin, 5-methoxytryptamine or tryptamine, we were able to achieve the simultaneous localization of two amines in glutaraldehyde-fixed sections of rat dorsal raphe nuclei. In this staining procedure, the first antigen was localized using 3,3'-diaminobenzidine (DAB), while the second antigen was stained using the 1-naphthol basic dye (2-NBD) method. The two antigens were localized in different cells or structures. No overlap of the staining was observed, thus indicating that dopamine is not localized with serotonin, 5-methoxytryptamine or tryptamine.

3,3'-Diaminobenzidine↗

Bipolarity of duodenal enterochromaffin cells in the rat.

Enterochromaffin cells of the rat duodenum have been studied immunocytochemically by use of a specific antiserum to serotonin. At the light-microscopic level serotonin immunoreactivity was observed in enterochromaffin cells located in the epithelium of the duodenal mucosa. Most of the serotonin-immunoreactive material was localized to the basal portion of the enterochromaffin cells, but small amounts of immunoreactive material were regularly observed in the apical portion. At the electron-microscopic level serotonin immunoreactivity in enterochromaffin cells was found to be concentrated over the dense cores of the cytoplasmic granules. The majority of these granules was located in the basal cytoplasm of the enterochromaffin cells, but serotonin-immunoreactive granules were also observed in the apical cytoplasm immediately beneath the microvilli. These observations indicate that duodenal enterochromaffin cells are bipolar and that they secrete serotonin both basally, to the circulation, and apically, to the gut lumen. Rat duodenal enterochromaffin cells thus appear to have an exocrine as well as an endocrine function.

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