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Biomedical subjects

M Geffard

Publications and source records attributed to M Geffard.

At least 145 records · Page 8Linked to original sources
PubMed1987

[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↗
PubMed1987

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↗
PubMed1987

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.

Animals↗
PubMed1987

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↗
PubMed1987

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.

Animals↗
PubMed1987

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.

Animals↗
PubMed1987

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.

Animals↗
PubMed1987

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.

Animals↗
PubMed1987

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↗
PubMed1987

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.

Animals↗
PubMed1987

Central GABAergic innervation of the pituitary in goldfish: a radioautographic and immunocytochemical study at the electron microscope level.

The GABAergic innervation of the goldfish pituitary was studied at the light and electron microscope levels by means of radioautography after in vitro incubation in tritiated gamma-aminobutyric acid (GABA) and immunocytochemistry using antibodies against GABA. Following incubation of pituitary fragments in a medium containing tritiated GABA, a selective uptake of the tracer was observed within the digitations of the neurohypophysis. Silver grain clusters were also observed in the adenohypophyseal tissue. At the electron microscope level, this uptake was found to correspond to nerve endings containing small clear and dense-core vesicles. These labeled profiles were located mainly in neurohypophyseal digitations in close apposition with the basement membrane separating the neurohypophysis from the adenohypophysis. However, they were also encountered in direct contact with most adenohypophyseal cell types in the different lobes. These results were confirmed by immunocytochemical data demonstrating the presence of numerous GABA immunoreactive fibers in both anterior and neurointermediate lobes. They were found either in the digitations of the neurohypophysis or in the adenohypophysis in direct contact with the glandular cells with a distribution and an ultrastructural aspect similar to those observed by radioautography. These data demonstrate that the pituitary of teleosts receives a massive GABAergic innervation. Although physiological data providing a functional significance for such an innervation are lacking, the present study suggests that, as already documented in mammals, GABA may be involved in the neuroendocrine regulation of pituitary functions in teleosts.

Animals↗
PubMed1987

Ultrastructural localization of GABA in the supraoptic nucleus and neural lobe.

Antibodies directed against the neurotransmitter gamma-aminobutyric acid (GABA) enabled the ultrastructural localization of GABA in conventional glutaraldehyde fixed and osmium postfixed material of the rat supraoptic nucleus and neural lobe. GABA was visualized using immunogold postembedding staining in axonal profiles that terminate on dendrites, axons or cell bodies throughout the supraoptic nucleus. The optimum ultrastructural preservation made possible the visualization of GABA terminals, also in the neural lobe. Here GABA axons were found to terminate synaptically on pituicytes and axonal profiles containing large dense core vesicles. These results emphasize, from an anatomical point of view, the potency of GABA to influence, as a transmitter, the release of vasopressin and oxytocin, both at the level of the cell body and of the neural lobe.

Animals↗
PubMed1987

Anatomical distribution and ultrastructural organization of the GABAergic system in the rat spinal cord. An immunocytochemical study using anti-GABA antibodies.

gamma-Aminobutyric acid (GABA)-containing elements have been studied by light and electron microscopy in the rat spinal cord, using immunocytochemistry with anti-GABA antibodies. Light microscopy showed immunoreactive somata localized principally in laminae I-III, and occasionally in the deeper laminae of the dorsal horn and in the ventral horn. Small somata were also observed around the central canal. Punctate GABA-immunoreactive profiles were particularly concentrated in laminae I-III, and moderately abundant in the deeper laminae and in the ventral horn where they were observed surrounding the unlabelled motoneurons. At the ultrastructural level, the punctate profiles corresponded to GABA-containing axonal varicosities or small dendrites. GABA-immunoreactive varicosities were presynaptic to labelled or unlabelled dendrites and cell bodies. Some unlabelled terminals presynaptic to unlabelled dendrites received symmetrical synaptic contacts from GABA-immunoreactive terminals. These results confirm data obtained with L-glutamate decarboxylase immunocytochemistry, and support the role of GABA in pre- and postsynaptic inhibition in the spinal cord, respectively via axoaxonal and axosomatic or axodendritic synapses.

Animals↗
PubMed1987

Immunocytochemical localization of dopamine in the prefrontal cortex of the rat at the light and electron microscopical level.

In the present study the dopaminergic innervation of the prefrontal cortex was studied by means of a recently developed anti-dopamine serum. This method can demonstrate endogenous dopamine in a specific way, and offers the opportunity to study the distribution of dopaminergic fibres in the cortex in detail in counterstained sections. Furthermore, dopaminergic nerve endings can be visualized at the electron microscopic level. Light microscopic observations demonstrated that the highest density of dopaminergic fibres in the frontal cortex is found in the prefrontal cortex and the infralimbic cortex. Within the prefrontal cortex, a good correlation is found between regional differences in distribution of dopaminergic fibres and the cytoarchitectonic parcellation of this part of the cortex. Outside the prefrontal cortex dopaminergic fibres were observed in adjacent frontal areas, the cortex surrounding the entire rhinal sulcus and the retrosplenial cortex. Electron microscopic observations demonstrated dopaminergic terminals through all cortical layers. The majority of dopaminergic terminals in the prefrontal cortex from synaptic contacts with dendritic processes. The synaptic profiles were usually symmetric and were characterized by the presence of many clear vesicles and an occasional dense-core vesicle.

Animals↗
PubMed1987

Monoclonal antibodies against glutaraldehyde-conjugated dopamine.

Four mice were immunized with dopamine (DA)-glutaraldehyde (G)--protein conjugates over a period of 8-10 weeks. Polyclonal antisera, obtained at various intervals, were tested using an enzyme-linked immunosorbent assay (ELISA). All had anti-conjugated DA antibodies. As soon as good antibody affinity was detected between 10(-10) and 10(-6) M, the mouse yielding the highest apparent affinity was killed, and the spleen was dissected out. Hybridomas were obtained from spleen cells fused with SP2/O/Ag myeloma cells. Supernatant culture media of hybridomas were tested for the presence of anti-conjugated DA antibodies with the ELISA method. Selected hybridomas giving good antibody affinity and specificity were then cloned by the limiting dilution technique. The resulting supernatant culture media were again tested by ELISA. Clones that gave a high antibody affinity (10(-10)-10(-8)M) for G-conjugated DA were used for histochemical localization of DA in rat brain. G-fixed rat brains were sectioned from the telencephalon to the mesencephalon, reduced with sodium borohydride, and prepared for peroxidase-antiperoxidase immunocytochemistry using supernatant (diluted 1:100) or ascites fluid (diluted 1:50,000). Dense networks of very fine fibers were observed in the striatum, septum, and cortex. Numerous immunoreactive cell bodies were found in the ventral tegmental area, the substantia nigra, the hypothalamus, and the dorsal raphe. The ELISA tests and adsorption controls suggested that the monoclonal antibody allowed highly specific detection of DA in tissues.

Aldehydes↗
PubMed1987

Specific antisera against the catecholamines: L-3,4-dihydroxyphenylalanine, dopamine, noradrenaline, and octopamine tested by an enzyme-linked immunosorbent assay.

Antisera were raised against L-3,4-dihydroxyphenylalanine (L-DOPA), dopamine (DA), noradrenaline (NA), and octopamine (OA). This was achieved by coupling each molecule to bovine serum albumin or human serum albumin using glutaraldehyde. The conjugated aromatic amines were kept in a reducing medium containing sodium metabisulfite. Antiserum specificity was tested using an enzyme-linked immunosorbent assay method for catecholamines. Competition experiments were done between the immunogen coated on the well plates and each catecholamine, either in the free state or in conjugated form, previously incubated with an antiserum. In each case, the nonconjugated compound was poorly recognized. The nonreduced conjugates of L-DOPA and DA were well recognized, whereas those of NA and OA were poorly immunoreactive. The cross-reactivity ratios established in the competition experiments allowed the specificity of the immune response to be defined. In each case, it was found to be high. The results suggest that the antibodies of L-DOPA and DA antisera recognize preferentially the catechol moiety, whereas for the anti-NA and anti-OA antibodies, the lateral chain is important.

Animals↗
PubMed1987

Anti-acetylcholine antibodies and the pathogenesis of myasthenia gravis.

Using an ELISA system, antibodies recognizing conjugated acetylcholine (ACh) were detected in sera of patients suffering from myasthenia gravis. The mean antibody level was three times higher in sera from myasthenic than from control patients. No correlation was found between anti-ACh antibody levels and anti-ACh receptor (AChR) titer. Also, the anti-ACh antibody titers were independent of sex and age of patients. Competition experiments demonstrated that the most immunoreactive compounds were choline-glutaryl-bovine serum albumin (BSA) and choline-succinyl-BSA. Antibodies present in the sera of myasthenic patients recognized an antigenic determinant mimicking conjugated ACh. The antibody affinity and specificity were sufficiently high for the detection of ACh in locust brain.

Acetylcholine↗