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M Geffard

Publications and source records attributed to M Geffard.

At least 91 records · Page 5Linked to original sources

Serotonergic innervation of the dorsal horn of rat spinal cord: light and electron microscopic immunocytochemical study.

The ultrastructure of serotonergic projections to the dorsal horn of the rat spinal cord has been investigated, using a highly specific polyclonal antiserum. The highest concentrations of immunoreactive profiles were found in lamina I and the outer part of lamina II (IIo). Intermediate concentrations were found in laminae III and IV, while the inner part of lamina II (IIi) was almost devoid of immunoreactivity. Whereas 60% of the profiles show at least one varicosity studded with synaptic vesicles, only one-fifth of the latter contributes to classical synapses, the remaining profiles being devoid of a facing postsynaptic density. The results are compared with those in the literature and our own results relative to other regions of the cord. It is concluded that the pauci-synaptic projections to the dorsal horn could correspond to a diffuse influence of serotonin, the targets for which are determined by the corresponding serotonergic receptors.

Animals↗

The metabolism of exogenous L-dopa in the brain: an immunohistochemical study of its conversion to dopamine in non-catecholaminergic cells of the rat brain.

The characterization and localization of non-catecholaminergic cells producing dopamine after L-Dopa load have been investigated in the normal rat brain by a direct immunohistochemical labelling of amines using specific antibodies. The detection of dopamine-containing non-catecholaminergic cells has been achieved in rats given a commonly used mixture of L-Dopa plus peripheral decarboxylase inhibitor, and compared to controls. Results indicate that serotoninergic neurons tend toward a switch of their metabolism into dopamine production after L-Dopa load in a dose-dependent manner. In addition small non-aminergic cells, identified as aromatic amino-acid decarboxylase-containing cells, were observed to produce dopamine after exogenous L-Dopa load. Possible implications of such results concerning the mode of action of L-Dopa in the brain are discussed.

Administration, Oral↗

Increase of auto anti-phosphatidylinositol antibodies in plasma of female rats during the appearance of DMBA-induced malignant mammary tumors.

Using a lipid-adapted ELISA, antiphosphatidylinositol (PtdIns) antibodies (Ab) have been found in sera of cancer patients. With the same procedure, we have checked their possible raising in plasma of female rats during the appearance of 7,12-dimethylbenz[a]anthracene (DMBA)-induced mammary tumors. Twenty days after DMBA administration, the mean anti-PtdIns Ab level was already higher than that of control rats (oil). Immunochemical analysis of the anti-PtdIns Ab site showed a rather high Ab avidity (8 x 10(-9) M, at half-displacement) and high specificity for glyceryl phosphate inositol residues. Other phospholipids (PL) were not recognized by the anti-PtdIns Ab induced by malignant cell transformation.

9,10-Dimethyl-1,2-benzanthracene↗

Noradrenaline-like terminals in the cat nucleus ventralis posterior of the thalamus.

Noradrenaline-like immunoreactivity in the cat nucleus ventralis posterior of the thalamus was investigated using an indirect immunocytochemical technique. Specific antinoradrenaline antibodies, raised in rabbits, were used. It was first verified that these antibodies recognize noradrenaline cells bodies of the locus coeruleus and their ascending axons in the ascending noradrenergic tract. In the nucleus ventralis posterior itself, noradrenaline-like fibers were observed. They were either randomly distributed or grouped around nonlabeled cell bodies. These neurons were generally oblong and measured 60-80 microns. With electron microscopy, preliminary results showed immunoreactive fibers in close apposition to unlabeled cell bodies or dendrites. The precise nature of these profiles was sometimes difficult to ascertain, since experiments were done in presence of detergent. In some cases symmetric synapses might be observed between immunoreactive axon terminals and unlabeled dendrites. The specificity of the reaction is discussed in the light of several control experiments.

Animals↗

Distribution of GABA immunoreactive systems in the forebrain and midbrain of the chameleon.

An immunocytochemical method, using glutaraldehyde fixation and an antiserum developed against a GABA-glutaraldehyde protein conjugate, permitted direct visualization of GABAergic structures in the brain of a reptile (chameleon). GABA immunoreactive cell bodies and nerve terminals were observed to be evenly distributed throughout the forebrain and midbrain. In the forebrain, GABA-positive perikarya were shown in all cortical areas, the septal area, the striatum, the dorsal ventricular ridge, and in the nucleus accumbens. In the midbrain, the optic tectum contained a dense and laminar distribution of GABA neurons. These neurons were also observed in the lateral geniculate nucleus, nucleus profundus mesencephali, nucleus opticus tegmenti and substantia nigra. Immunoreactive nerve fibers and terminals were observed in the same structures and, additionally, in the tractus septo-hypothalamicus, habenula complex, median eminence, intermediate lobe of the pituitary, basal part of the subcommissural organ, torus semicircularis and nucleus reticularis isthmi. These results provide a framework for a further electron microscopic analysis of the GABAergic innervation of some encephalic areas involved in physiological regulations particular to this species especially the visual system.

Animals↗

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↗