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M Geffard

Publications and source records attributed to M Geffard.

At least 91 records · Page 5Linked to original sources

Monoaminergic interaction in the central nervous system: a morphological analysis in the locus coeruleus of the rat.

The locus coeruleus of the rat is richly innervated by many aminergic neurons varying in amine content and in site of origin. There are adrenergic and noradrenergic neurons originating in the medulla oblongata, dopaminergic from the hypothalamus, serotonergic from the mesencephalon and also intrinsic noradrenergic neurons in the locus coeruleus complex. Of these, adrenergic and dopaminergic inputs appear relatively specific and powerful.

Animals↗

Polyclonal anti-idiotypic antibodies as internal images of dopamine. Applications for biochemical and morphological studies of DA receptors in the rat brain.

Polyclonal anti-idiotypic antiserum raised against both rabbit and monoclonal anti-dopamine (DA) antibodies was produced in rabbits. It was characterized for its specificity and was shown to (1) inhibit the binding of both polyclonal and monoclonal idiotypic anti-DA antibodies directed to immobilized DA conjugates; (2) inhibit the binding of (3H) DA to rat brain membranes; (3) to cross-react with a peptide extracted from a neuroblastoma cell line (NCB-20), known to express functional DA receptors. Finally, immunocytochemical studies were performed on paraformaldehyde-fixed rat brain. Anti-idiotypic antibodies were used to visualize the cellular and subcellular distribution of DA receptor binding sites in the striatum, a region that contains both D1 and D2 receptors subtypes. Under the electron microscope, the immune reaction product was observed to be concentrated in postsynaptic sites belonging mainly to dendritic spines, while presynaptic structures were sparsely labeled.

Animals↗

Locomotor bias produced by intra-accumbens and intracaudate injection of polyclonal dopamine anti-idiotypic antibodies.

The preceding article described anti-idiotypic antibodies to conjugated dopamine (AIDA); results were consistent with the hypothesis that these antibodies contained the internal image of conjugated dopamine (DA-G-BSA) and binded to dopamine (DA) receptors. We further tested these anti-idiotypic antibodies to conjugated dopamine by examining the functional changes produced by unilateral injection of AIDA (or DA-G-BSA) into the nucleus accumbens or into the medio-dorsal caudate in mice. Our results showed that unilateral injection of AIDA (or DA-G-BSA) into the nucleus accumbens or into the medio-dorsal caudate produced an ipsilateral locomotor asymmetry in amphetamine-treated animals which was similar to the one produced by unilateral intra-caudate injection of haloperidol (a non-specific DA antagonist). The asymmetry was site specific: injection of AIDA around the nucleus accumbens or into the anterior caudate was not effective. The asymmetry was also shown not to depend on the peripheral injection of amphetamine. Taken together, the present results and those in the preceding article suggest that the AIDA contains the internal image of DA-G-BSA and that these two substances bind to DA receptors to produce behavioral changes.

Animals↗

Autoantibodies directed against lipid membrane components in sera of patients with malignant tumors.

Tumor-associated antigens (Ag) are expressed on neoplastic cells. Using an enzyme-linked immunosorbent assay (ELISA), an attempt was made to evaluate the autoimmune responses directed against lipid membrane components. A comparison was made of autoantibody (autoAb) levels in human sera of 684 patients with malignant tumors and those of 185 controls (healthy subjects and patients suffering from other diseases). A highly significant difference was found between the immunological binding of the groups for only one phospholipids (PL), i.e., phosphatidylinositol (PI). Using ELISA tests and PI-related compounds differing in fatty acid residue types and/or in phosphatidyl group, it was demonstrated that the hydrophilic residue is the immunodominant part recognized by the autoAb detected in sera of cancer patients.

Adult↗

Expression of GABA-immunoreactivity by spinal motoneurons of some vertebrates.

Electrophysiological and biochemical investigations have shown that gamma-aminobutyric acid (GABA) is the main inhibitory neurotransmitter in the vertebrate central nervous system. However, the present study shows that some motoneurons located in the spinal cord of young chickens and adult monkeys display a GABA-like immunoreactivity. The expression of GABA immunoreactivity in vertebrate motoneurons suggests that this inhibitory amino acid is colocalized with acetylcholine and could play a role in the neuromuscular transmission.

Animals↗

Dopamine- and dopa-immunoreactive neurons in the cat forebrain with reference to tyrosine hydroxylase-immunohistochemistry.

The distribution of cell bodies containing immunoreactivities to dopamine (DA), L-3,4-dihydroxyphenylalanine (DOPA) and tyrosine hydroxylase (TH) was studied immunohistochemically in the cat forebrain especially in the hypothalamus with or without intraventricular administration of colchicine. In normal cats, DA-immunoreactive (IR) neurons, whose intensity of immunostainings was variable from one to another, were localized exclusively in the hypothalamus and showed a distribution pattern similar to that of TH-IR ones. They were distributed in the posterior, dorsal and periventricular hypothalamic areas. Arcuate cells showed no or very weak DA-immunoreactivity. Weak to intense DOPA-IR cells were distributed in a similar manner to DA-IR ones but were far smaller in number. In colchicine-treated animals, DA- and DOPA-immunoreactivities were enhanced particularly in arcuate and dorsal hypothalamic cells. A cluster composed of small DA- and DOPA-IR cells was identified in the area ventral to the mamillothalamic tract equivalent to rat A13c TH-IR cell group. Colchicine treatment enabled us to visualize a large number of TH-IR perikarya in the medial and lateral preoptic areas, anterior commissure nucleus, basal forebrain, area closely related to the organum vasculosum laminae terminalis, and some in the bed nucleus of the stria terminalis as has been reported in other species. However, virtually none of these cells contained detectable DA- and DOPA-immunoreactivities.

Animals↗

Immunohistochemical evidence for GABAergic cell bodies in the medial nucleus of the trapezoid body and in the lateral vestibular nucleus in the guinea pig brainstem.

The presence of gamma-aminobutyric acid (GABA) in two brainstem nuclei is demonstrated by using a pre-embedding immunohistochemical procedure followed by staining intensification. Firstly, immunoreactivity was found in numerous cell bodies and profiles of the medial nucleus of the trapezoid body (MNTB). Secondly, numerous neurons including giant Deiters' cells, terminals and fibers were strongly labelled within the lateral vestibular nucleus (LVN). These observations suggest that the inhibitory part of the efferent innervation of outer hair cells in the cochlea can originate from the MNTB, and that GABAergic neurons in the LVN may contribute to information processing within this nucleus.

Animals↗

Simultaneous detection of tryptamine and dopamine in rat substantia nigra and raphe nuclei using specific antibodies.

Using a double-labelling procedure, morphological relationships existing between dopaminergic and indoleaminergic neuronal systems in the rat brain were investigated. First, thanks to a tryptamine (T) antiserum, we visualized this indoleamine in all mesencephalic regions and showed that the T-immunoreactivity (IR) seems to overlap with the stainings observed from serotonin and 5-methoxytryptamine antisera. Secondly, using a monoclonal anti-dopamine (DA) antibody and our anti-T antibodies, the simultaneous and specific detection of these compounds enabled us to define the spatial relationships existing between the dopaminergic and tryptaminergic neuronal systems from the substantia nigra (SN) to the raphe nuclei. No co-localization existed, but the intensity of T-IR decreased from back to front, whereas the DA-staining decreased in the opposite way, indicating possible interactions at the end of the SN and the B9 area.

Animals↗

Immunocytochemical and autoradiographic studies of the endocrine cells interacting with GABA in the rat stomach.

There are now increasing evidences suggesting that GABA is able of direct interaction with certain endocrine cells. In the present study, highly specific anti-GABA-glutaraldehyde antibodies and 3H-GABA uptake were used at the light and electron microscope levels to investigate the occurrence of cells containing endogenous GABA or taking up exogenous GABA in the mucosal antrum and corpus of the rat stomach. Only certain endocrine cell types of both regions were immunostained or grain-labelled. However, the morphology of their secretory granules did not allow to identify the nature of their hormone with certainty but suggested that somatostatin-like cells could interact with GABA. The combination of gastrin and somatostatin immunodetection with 3H-GABA uptake autoradiography at the light microscope level, revealed that a subpopulation of somatostatin-like cells and other still unidentified endocrine cells are able to take up GABA, while the gastrin-like cells are not. These results reinforce the hypothesis that certain endocrine cell types of the diffuse endocrine system of the digestive tract are able to directly interact with GABA.

APUD Cells↗

Immunohistochemistry of endogenous L-DOPA in the rat posterior hypothalamus.

The aim of this work was to study L-DOPA-containing neuronal structures of the rat posterior and dorsal hypothalamus by means of immunohistochemistry using antiserum against glutaraldehyde conjugated L-DOPA. Aspects and distribution of L-DOPA immunoreaction among cells of the supramammillary nucleus and the A11, A13c and A13 cell groups are described and compared to dopamine immunoreactivity, mainly through a double colored labelling procedure employing a color modification of the DAB reaction by metallic ions. Differences between L-DOPA and dopamine stainings within cell groups as the presence of cells with predominant or exclusive L-DOPA coloration are tentatively explained under the light of previous findings using immunohistochemistry of catecholamines synthesizing enzymes and catecholamines histofluorescence.

Animals↗

Noradrenaline axon terminals in adult rat neocortex: an immunocytochemical analysis in serial thin sections.

Peroxidase-antiperoxidase electron microscope immunocytochemistry with an antiserum against noradrenaline-glutaraldehyde-protein conjugate was used to identify cortical noradrenaline terminals (axonal varicosities) from the upper layers of the frontal, parietal and occipital cortex in adult rat. A large number of immunostained varicosities were examined in serial thin sections, and compared with a control population of randomly chosen unlabeled terminals from the same sections. Both groups of varicosities were measured and scrutinized for the presence of a junctional complex indicative of synaptic specialization. Cellular elements juxtaposed to the membrane of both types of varicosities were also identified and counted. Noradrenaline varicosities in all three cortical regions averaged 0.65 microns in diameter. In contrast to their unlabeled counterparts, these profiles rarely showed a membrane differentiation characteristic of a synaptic contact (junctional complex). The rare junctional complexes formed by cortical noradrenaline varicosities were invariably symmetrical and almost always found on dendritic shafts. The microenvironment of noradrenaline varicosities also differed, exhibiting a greater number of apposed axonal varicosities and a smaller number of dendritic spines than that of the random population. The proportion of noradrenaline varicosities making a synaptic contact (synaptic incidence) was determined by plotting the incidence of visible junctions as a function of the number of thin sections available for examination. As extrapolated for whole varicosities after linear transformation (double reciprocal plot), this proportion was 17% or 26% depending on the stringency of the criteria used in identifying the junctional complex. The same analysis provided a figure of 98% for the control population. The present study largely confirmed our initial radioautographic characterization of the cortical noradrenaline innervation as a mostly non-junctional system, and also indicated that these varicosities are set in a particular microenvironment. These new data further support the eventuality of a diffuse release of cortical noradrenaline in the extracellular space, compatible with both its neuromodulatory role and multiplicity of actions on diverse cellular targets in the cerebral cortex. The functions assigned to the coeruleocortical noradrenaline system must therefore be viewed as the product of a widespread and ubiquitously distributed neuronal organization characterized by loose intercellular relationships. This system might be capable of selectivity and specificity of action, however, owing to the distribution of its receptors, and in view of intrinsically or extrinsically driven control mechanisms triggered by the release of its own or other transmitters and which may also involve target-initiated feedback mechanisms.

Animals↗

Neurons containing messenger RNA encoding glutamate decarboxylase in rat hypothalamus demonstrated by in situ hybridization, with special emphasis on cell groups in medial preoptic area, anterior hypothalamic area and dorsomedial hypothalamic nucleus.

Previous deafferentation studies have suggested that most hypothalamic GABAergic innervation originates from neurons within the hypothalamus. We have investigated the distribution of GABAergic cell groups in the rat hypothalamus by means of the in situ hybridization technique, using a cDNA probe for messenger RNA encoding glutamate decarboxylase. Several major GABAergic cell groups were demonstrated, including cells of the tuberomammillary nucleus, arcuate nucleus, suprachiasmatic nucleus, medial preoptic area, anterior hypothalamic area, the dorsomedial hypothalamic nucleus, perifornical area, and lateral hypothalamic area. The most prominent glutamate decarboxylase mRNA-containing cell groups were located in the medial preoptic area, anterior hypothalamic area and dorsomedial hypothalamic nucleus, and were composed of small- to medium-sized neurons. Compared to previously well-characterized GABAergic cell groups in the tuberomammillary nucleus, reticular thalamic nucleus, and non-pyramidal cells of cerebral cortex, the cells of these GABAergic groups demonstrated only weak cDNA labelling, indicating that they contain lower levels of glutamate decarboxylase mRNA. Several types of control experiments supported the specificity of this cDNA labelling, and the GABAergic nature of these cell populations was further supported by detection of glutamate decarboxylase and GABA immunoreactivity. Abundance of GABAergic cells in many hypothalamic nuclei indicates that GABA represents quantitatively the most important transmitter of hypothalamic neurons, and may be involved in neuroendocrine and autonomic regulatory functions.

Animals↗

Dissimilar responses of adult thalamic monoaminergic and somatosensory afferent fibers to implantation of thalamic fetal cells.

It is generally accepted that transplanted fetal neurons can, after several weeks to months, establish connections with the host CNS. Host afferent systems seem, however, to show different types of responses to the presence of grafted fetal neurons. The present study is a preliminary step to identify mechanisms involved in the reactions of adult axons to transplanted fetal neurons. The right ventrobasal thalamus of adult rats was depleted of neurons by in-situ injection of kainic acid and cell suspensions from homotopic thalamic embryonic primordia which were injected into the lesioned area. After various post-implantation delays, ranging from five to 30 days, two types of experiments were performed: (i) noradrenaline and serotonin immunohistochemistry with specific antibodies on alternate sections; and (ii) anterograde tracing using wheat germ agglutinin conjugated to horseradish peroxidase from the dorsal column nuclei and the principal sensory trigeminal nucleus. Five days after transplantation, host monoaminergic fibers (either noradrenergic or serotoninergic) had already grown into the transplants. Ingrowing fibers were thin and poorly varicose, exhibiting endings morphologically similar to the growth cones observed during axogenesis. Seven days after grafting, growth cones were no longer visible and monoaminergic fibers exhibited either normal-sized or very large varicosities. Large varicosities progressively decreased in number and, after three weeks, the fibers displayed a normal adult morphology, forming a dense network all over the transplants. In contrast, host somatosensory afferents, labeled by anterograde transport of wheat germ agglutinin conjugated to horseradish peroxidase, did not grow into the transplants. Intermingling of somatosensory afferents and transplanted cells was observed only after 10 days, when grafted neurons extended outside the original transplantation site into the neuron-depleted area containing the somatosensory afferents. The present results demonstrate that adult monoaminergic and somatosensory afferents, when deprived of their usual target, do not react in a similar way to the addition of fetal neurons. It is proposed that adult monaminergic fibers have the ability to regain morphological (and probably functional) immature forms which were considered to be restricted to the period of axogenesis or to lesion-induced regeneration. In contrast, fetal transplants do not seem to induce, by themselves, a similar alteration of genetic expression in adult somatosensory neurons. It has been proposed that "diffuse" and "point-to-point" axonal systems may be differentiated in the CNS on anatomical bases. The present results add to the identification of two different systems by demonstrating that, in the thalamus, they present dissimilar responses to the implantation of fetal cells.

Animals↗

Monoclonal anti-conjugated azelaic acid antibody production: application to multiple sclerosis.

We have previously reported the existence of anti-conjugated azelaic acid (Aze A) antibodies in the serum of patients with multiple sclerosis (MS). In order to demonstrate the specificity of these antibodies, we have produced a monoclonal antibody directed against Aze A conjugated by an acylation reaction to a protein. In competition experiments, with ELISA method, we demonstrated that a part of the antibodies, raised in rabbit after immunization by human immunoglobulins (Ig) of MS patients, recognized the antigen-combining site of our monoclonal anti-conjugated Aze A antibody. These results clearly demonstrate that a part of human Ig obtained from sera of MS patients shared common idiotopes with mouse monoclonal antibody raised against conjugated Aze A.

Acylation↗

Existence of L-dopa immunoreactive neurons in the rat preoptic area and anterior hypothalamus.

We demonstrate the presence of L-DOPA (L-3,4-dihydroxyphenylalanine)-labelled cell bodies and fibers in the rat preoptic and anterior hypothalamic areas, using a rabbit polyclonal antiserum against conjugated L-DOPA and a glutaraldehyde coupled immunohistochemical technique. In contrast, using a monoclonal anti conjugated dopamine (DA) antibody, no DA labelled neurons were detectable in the preoptic region, whereas a few weakly immunostained DA neurons began to appear in the ventral part of the medial preoptic area. These observations point to the hypothesis that these cells contain mainly L-DOPA neurons in the preoptic and anterior hypothalamic areas. Thus, we suggest that L-DOPA plays a more important role in the hypothalamic functions than has been hitherto assumed.

Animals↗

The distribution of 5-HT immunoreactive systems in the brain of a saurian, the chameleon.

The distribution of serotonin immunoreactive cell bodies and fibers was studied in the chameleon brain by using the immunohistochemical technique with antisera against serotonin coupled to a carrier with glutaraldehyde. Serotonin perikarya were found in the caudal midbrain tegmentum, in the lateral part of the nucleus reticularis isthmi, the lateral part of the nucleus interpeduncularis and along the midline in the raphe superior. More caudally, the serotonin immunoreactive cell bodies were located along the nucleus raphe inferior and ventrolaterally in the vicinity of the olivary complex. No immunoreactive cell bodies were found in the spinal cord nor in the paraventricular organ (PVO) of the hypothalamus. Immunoreactive fibers were observed in the entire brain. Prominent concentrations were found in the dorsal cortex, lateral septum, lateral geniculate nucleus, median eminence, pretectal nucleus, nucleus interpeduncularis, vestibular nucleus and olivary complex. Descending serotonin immunoreactive fibers were found in particular in the ventral motoneuron area in the spinal cord. One of the most interesting findings in this study was the lack of immunoreactive CSF contacting neurons in the PVO and the observation of an extensive plexus of supraependymal fibers, a feature reported so far only in mammals.

Animals↗

Co-localization of tyrosine hydroxylase and GABA immunoreactivities in human cortical neurons.

Samples of human cerebral cortex were stained immunocytochemically for tyrosine hydroxylase (TH) and gamma-aminobutyric acid (GABA). TH-positive neurons were in small number and predominated in the deep infragranular layers V-VI contrasting with numerous GABA-positive neurons scattered in all layers. Co-localization of TH- and GABA-like immunoreactivities in a single cell was studied by the double immunolabeling technique with the elution-restaining procedure. Only 50% of the TH-positive neurons also expressed GABA-like immunoreactivities. The two markers were detectable in the somata and not in the processes of the cells. The double-labeled cells were mainly fusiform and medium-sized and were observed in layer VI. These observations suggest that the TH-positive cells form a mixed neuronal population, only a part of which corresponds to the GABAergic class of intrinsic interneurons.

Cerebral Cortex↗

Endogenous L-dopa in the rat dorsal vagal complex: an immunocytochemical study by light and electron microscopy.

The aim of this work was to examine L-DOPA immunoreactivity (L-DOPA-IR) in the dorsal vagal complex (DVC) of the rat medulla oblongata containing A2/C2 catecholaminergic cell groups, in order to further evaluate the previously proposed hypothesis that various pools of endogenous L-DOPA could be immunocytochemically demonstrated in the mammalian brain. For this purpose, L-DOPA-IR was studied in DVC in comparison with both some other catecholaminergic areas and dopamine immunoreactivity (DA-IR) on adjacent sections of the same brain, by using specific antibodies against glutaraldehyde conjugated L-DOPA and DA. Also, the first preliminary observations of L-DOPA-IR in DVC neurons at the ultrastructural level are reported. The following main results were obtained: (1) bright, intense and homogeneous L-DOPA staining was found in perikarya and proximal neuronal processes situated within the rostrocaudal extension of the DVC; (2) this staining pattern was readily distinct from weak and heterogeneous DA staining; (3) an inverse L-DOPA/DA staining pattern ratio was identified between the DVC and the mesencephalon; (4) L-DOPA-IR at electron microscopic level was roughly similar to that previously observed for DA-IR in mesencephalic cells and their presumptive projections. Although some discrepancies were noticed between L-DOPA staining and data from the literature on tyrosine hydroxylase labeling, our results could not invalidate the hypothesis that, among high L-DOPA/DA ratio containing neurons, some cells in the DVC may contain only L-DOPA.

Animals↗