RESUMO
OBJECTIVE: Depression is accompanied by activation of immuno-inflammatory and oxidative and nitrosative stress (IO&NS) pathways, and increased IgM/IgA responses to lipopolysaccharide (LPS) of gram-negative commensal bacteria. The latter suggests that bacterial translocation has caused IgM/IgA responses directed against LPS. Bacterial translocation may drive IO&NS responses. METHOD: To examine the associations between IgM/IgA responses to LPS and IO&NS measurements, including plasma/serum interleukin-1 (IL-1), tumor necrosis factor (TNF)α, neopterin, lysozyme, oxidized LDL (oxLDL) antibodies, peroxides, and IgM (auto)immune responses against malondialdehyde (MDA), azelaic acid, phophatidyl inositol (Pi), NO-tryptophan and NO-tyrosine in depressed patients and controls. RESULTS: We found significant positive associations between IgM/IgA responses to LPS and oxLDL antibodies, IgM responses against MDA, azelaic acid, Pi, NO-tryptophan, and NO-tyrosine. The IgA responses to LPS were correlated with lysozyme. There were no significant positive correlations between the IgM/IgA responses to LPS and IL-1 and neopterin. CONCLUSION: The findings show that in depression there is an association between increased bacterial translocation and lysozyme production, an antibacterial compound, O&NS processes, and autoimmune responses directed against O&NS generated neoantigenic determinants. It is suggested that bacterial translocation may drive IO&NS pathways in depression and thus play a role in its pathophysiology.
Assuntos
Autoimunidade/imunologia , Translocação Bacteriana/imunologia , Transtorno Depressivo Maior/imunologia , Epitopos/imunologia , Inflamação/etiologia , Sistema Nervoso/imunologia , Estresse Oxidativo/fisiologia , Adulto , Autoimunidade/fisiologia , Estudos de Casos e Controles , Estudos Transversais , Transtorno Depressivo Maior/sangue , Transtorno Depressivo Maior/fisiopatologia , Feminino , Humanos , Imunoglobulina A/imunologia , Imunoglobulina M/imunologia , Inflamação/imunologia , Interleucina-1/sangue , Lipopolissacarídeos/imunologia , Masculino , Muramidase/sangue , Neopterina/sangue , Sistema Nervoso/fisiopatologia , Fator de Necrose Tumoral alfa/sangueRESUMO
A specific antibody to acetylcholine was raised and used as a marker for cholinergic neurons in the rat central nervous system. The acetylcholine conjugate was obtained by a two-step immunogen synthesis procedure. An enzyme-linked immunosorbent assay was used to test the specificity and affinity of the antibody in vitro; the results indicated high affinity. A chemical perfusion mixture of allyl alcohol and glutaraldehyde was used to fix the acetylcholine in the nervous tissue. Peroxidase-antiperoxidase immunocytochemistry showed many acetylcholine-immunoreactive cells and fibers in sections from the medial septum region.
Assuntos
Acetilcolina/imunologia , Encéfalo/anatomia & histologia , Fibras Colinérgicas/anatomia & histologia , Acetilcolina/metabolismo , Animais , Especificidade de Anticorpos , Mapeamento Encefálico , Técnicas Imunoenzimáticas , Masculino , RatosRESUMO
Using highly specific antisera directed against conjugated d-amino acids, the distribution of d-glutamate-, d-tryptophan-, d-cysteine-, d-tyrosine- and d-methionine-immunoreactive structures in the rat brain was studied. Cell bodies containing d-glutamate, but not d-glutamate-immunoreactive fibers, were found. Perikarya containing this d-amino acid were only found in the mesencephalon and thalamus of the rat CNS. Thus, the highest density of cell bodies containing d-glutamate was observed in the dorsal raphe nucleus, the ventral part of the mesencephalic central gray, the superior colliculus, above the posterior commissure, and in the subparafascicular thalamic nucleus. A moderate density of immunoreactive cell bodies was observed in the dorsal part of the mesencephalic central gray, above the rostral linear nucleus of the raphe, the nucleus of Darkschewitsch, and in the medial habenular nucleus, whereas a low density was found below the medial forebrain bundle and in the posterior thalamic nuclear group. Moreover, no immunoreactive fibers or cell bodies were visualized containing d-tryptophan, d-cysteine, d-tyrosine or d-methionine in the rat brain. The distribution of d-glutamate-immunoreactive cell bodies in the rat brain suggests that this d-amino acid could be involved in several physiological mechanisms. This work reports the first visualization and the morphological characteristics of conjugated d-glutamate-immunoreactive cell bodies in the rat CNS using an indirect immunoperoxidase technique. Our results suggest that the immunoreactive neurons observed have an uptake mechanism for d-glutamate.
Assuntos
Encéfalo/metabolismo , Ácido Glutâmico/metabolismo , Imunoquímica , Animais , Encéfalo/citologia , Mapeamento Encefálico , Ensaio de Imunoadsorção Enzimática/métodos , Masculino , Neurônios/citologia , Neurônios/metabolismo , Ratos , Ratos Sprague-DawleyRESUMO
It is known that kynurenic acid (KYNA) exerts a neuroprotective effect against the neuronal loss induced by ischemia; acting as a scavenger, and exerting antioxidant action. In order to study the distribution of KYNA, a highly specific monoclonal antibody directed against KYNA was developed. This distribution was studied in control rats and in animals in which a middle cerebral artery occlusion (stroke model) was induced. By double immunohistochemistry, astrocytes containing KYNA and GFAP were exclusively found in the ipsilateral cerebral cortex and/or striatum, at 2, 5 and 21days after the induction of stroke. In control animals and in the contralateral side of the stroke animals, no immunoreactivity for KYNA was found. Under pathological conditions, the presence of KYNA is reported for the first time in the mammalian brain from early phases of stroke. The distribution of KYNA matches perfectly with the infarcted regions suggesting that, in stroke, this overexpressed molecule could be involved in neuroprotective/scavenger/antioxidant mechanisms.
Assuntos
Córtex Cerebral/metabolismo , Corpo Estriado/metabolismo , Ácido Cinurênico/metabolismo , Neuroproteção , Acidente Vascular Cerebral/metabolismo , Regulação para Cima , Animais , Masculino , Ratos , Ratos Wistar , Distribuição TecidualRESUMO
The distribution of thiamine-immunoreactive structures was studied in the brain of the monkey using an indirect immunoperoxidase technique. Fibers containing thiamine, but no thiamine-immunoreactive cell bodies, were found. The highest density of fibers containing thiamine was observed in the pulvinar nucleus and in the region extending from the pulvinar nucleus to the caudate nucleus. In the mesencephalon, immunoreactive fibers containing thiamine were only found at rostral level close to the medial lemniscus (at the mesencephalic-diencephalic junction). In the thalamus, the distribution of thiamine-immunoreactive structures was more widespread. Thus, immunoreactive fibers were found in nuclei close to the midline (centrum medianum/parafascicular complex), in the ventrolateral thalamus (medial geniculate nucleus, inferior pulvinar nucleus), and in the dorsolateral thalamus (lateral posterior nucleus, pulvinar nucleus). Finally, in the anterior commissure and in the cerebral cortex a low density immunoreactive fibers was visualized. Thus, in the brainstem, no immunoreactive structures were visualized in the medulla oblongata, pons, or in the medial-caudal mesencephalon, and no immunoreactive fibers were observed in the cerebellum, hypothalamus and in the basal ganglia. The present report describes the first visualization and the morphological characteristics (thick, smooth and short, medium or long in length) of the thiamine-immunoreactive fibers in the primate central nervous system using an antiserum directed against this vitamin. The distribution of thiamine-immunoreactive structures in the monkey brain suggests that this vitamin could be involved in several physiological mechanisms.
Assuntos
Encéfalo/citologia , Fibras Nervosas/metabolismo , Fibras Nervosas/fisiologia , Tiamina/metabolismo , Tiamina/fisiologia , Animais , Especificidade de Anticorpos , Química Encefálica , Mapeamento Encefálico , Imuno-Histoquímica , Macaca fascicularis , Masculino , Mesencéfalo/metabolismoRESUMO
Using an antiserum directed against the vitamin riboflavin, we studied the distribution of riboflavin-like immunoreactive structures in the monkey brain. In the mesencephalon, at the level of the mesencephalic-diencephalic junction, single riboflavin-like immunoreactive fibers were observed in its dorsal part, whereas a low density of immunoreactive fibers was found below the surface of the section and close to substantia nigra, and a high density was observed above the substantia nigra and close to the medial geniculate nucleus. In the thalamus, single riboflavin-like immunoreactive fibers were found in the ventral regions of the lateral posterior and the medial geniculate nuclei; a low density in the region located above the medial and lateral geniculate nuclei and a high density in the ventral part of the pulvinar nucleus and in the region extending from this latter to the caudate nucleus. Immunoreactive fibers were not observed in the medulla oblongata, pons, cerebellum, hypothalamus, basal ganglia and cerebral cortex. Moreover, no riboflavin-like immunoreactive cell bodies were observed in the monkey brain. The distribution of riboflavin-like immunoreactive fibers in the monkey suggests that this vitamin could be involved in several physiological mechanisms.
Assuntos
Imuno-Histoquímica , Mesencéfalo/química , Riboflavina/análise , Tálamo/química , Animais , Imuno-Histoquímica/métodos , Macaca fascicularis , Masculino , Mesencéfalo/citologia , Fibras Nervosas/química , Reprodutibilidade dos Testes , Tálamo/citologiaRESUMO
High levels of anti-phosphatidylinositol (PtdIns) autoantibodies (autoAb) have been previously described in sera of cancer patients and in plasma of dimethylbenzanthracene-treated female Sprague-Dawley rats. The presence of anti-PtdIns autoAb was tested in a model of highly malignant sarcomas induced by a single dose of benzo(a)pyrene [B(a)P] diluted in sesame oil and injected in female Sprague-Dawley rats. Significantly elevated levels of anti-PtdIns autoAb were found in sera of B(a)P-treated rats 40 days after B(a)P administration, whereas no significant levels of anti-PtdIns autoAb were noted in oil- or benzo(e)pyrene-treated rats. After day 60, autoantibody levels plateaued in B(a)P-treated rats, and highly malignant sarcomas appeared with 100% efficiency around day 100. Anti-PtdIns autoAb avidity and specificity were found to be high.
Assuntos
Anticorpos Antineoplásicos/análise , Autoanticorpos/análise , Benzo(a)pireno , Neoplasias Experimentais/imunologia , Fosfatidilinositóis/imunologia , Animais , Autoanticorpos/imunologia , Benzopirenos , Divisão Celular/fisiologia , Transformação Celular Neoplásica/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Neoplasias Experimentais/induzido quimicamente , Neoplasias Experimentais/patologia , Valor Preditivo dos Testes , Ratos , Ratos Endogâmicos , Fatores de TempoRESUMO
Using an immunohistochemical technique, we have studied the distribution of 3-OH-anthranilic acid (3-HAA) in the rat brain. Our study was carried out in control animals and in rats in which a stroke model (single transient middle cerebral artery occlusion) was performed. A monoclonal antibody directed against 3-HAA was also developed. 3-HAA was exclusively observed in the infarcted regions (ipsilateral striatum/cerebral cortex), 2, 5 and 21 days after the induction of stroke. In control rats and in the contralateral side of the stroke animals, no immunoreactivity for 3-HAA was visualized. Under pathological conditions (from early phases of stroke), we reported for the first time the presence of 3-HAA in the mammalian brain. By double immunohistochemistry, the coexistence of 3-HAA and GFAP was observed in astrocytes. The distribution of 3-HAA matched perfectly with the infarcted regions. Our findings suggest that, in stroke, 3-HAA could be involved in the tissue damage observed in the infarcted regions, since it is well known that 3-HAA exerts cytotoxic effects.
Assuntos
Ácido 3-Hidroxiantranílico/metabolismo , Encéfalo/metabolismo , Acidente Vascular Cerebral/metabolismo , Animais , Encéfalo/patologia , Química Encefálica , Masculino , Ratos , Ratos Wistar , Acidente Vascular Cerebral/patologiaRESUMO
A highly specific monoclonal antibody directed against nitric oxide-tryptophan (NO-W) with good affinity (10-9 M) and specificity was developed. In the rat brain, using an indirect immunoperoxidase technique, cell bodies containing NO-W were exclusively found in the intermediate and dorsal parts of the lateral septal nucleus. No immunoreactive fibres were found in the rat brain. This work reports the first visualization and the morphological characteristics of cell bodies containing NO-W in the mammalian brain. The restricted distribution of NO-W in the rat brain suggests that this molecule could be involved in specific physiological mechanisms.
Assuntos
Óxido Nítrico/metabolismo , Núcleos Septais/metabolismo , Triptofano/análogos & derivados , Triptofano/metabolismo , Animais , Química Encefálica/fisiologia , Ratos , Ratos Wistar , Núcleos Septais/citologiaRESUMO
Immunoreactivity to p-tyramine, one of the natural trace amines, was studied in the rat brain by an anti-p-tyramine antibody. Immunoreactivity to this amine is very weak in the nigrostriatal dopaminergic neurons and terminals, and weak in the locus coeruleus noradrenergic ones. It was intensified in these structures after monoamine oxidase inhibition. On the other hand, this amine was highly concentrated in the median eminence of the mediobasal hypothalamus, in which its physiological function on prolactin release has been demonstrated.
Assuntos
Eminência Mediana/metabolismo , Tiramina/metabolismo , Animais , Descarboxilases de Aminoácido-L-Aromático/metabolismo , Dopamina/metabolismo , Imuno-Histoquímica/métodos , Masculino , Eminência Mediana/efeitos dos fármacos , Inibidores da Monoaminoxidase/farmacologia , Ratos , Substância Negra/metabolismoRESUMO
Antibodies against dopamine and p-tyramine were raised in rabbits. The two catecholamines were conjugated to albumin by glutaraldehyde. The specificity of the antibodies was established by equilibrium dialysis competition experiments using an immunoreactive tritiated derivative synthesized by coupling dopamine or p-tyramine to N-alpha-acetyl-L-lysine N-methylamide with glutaraldehyde. Hence, these radiolabelled ligands mimicked the antigenic determinant of conjugated immunogens. A comparison of the data obtained showed the high specificity of each antiserum for its hapten coupled by glutaraldehyde. The anti-dopamine antibodies recognized dopamine-glutaraldehyde but not p-tyramine-glutaraldehyde. The opposite occurred for the anti-p-tyramine antibodies. A slight modification of the molecular structure provided the opportunity for a specific response against that molecule. But this difference was more important when related to the hapten region where the antibody affinity was maximal. The cross-reactivity was observed to be more important dopamine and p-tyramine than between dopamine and noradrenaline on the one hand and between p-tyramine and dopamine than p-tyramine and octopamine on the other hand.
Assuntos
Anticorpos/imunologia , Catecolaminas/imunologia , Dopamina/imunologia , Tiramina/imunologia , Animais , Afinidade de Anticorpos , Especificidade de Anticorpos , Levodopa/imunologia , Norepinefrina/imunologia , Octopamina/imunologia , Coelhos , Ensaio Radioligante , Fatores de TempoRESUMO
At the late gastrula-early neurula stage some embryonic neuroblasts from neural plate and neural fold present apparently as a consequence of neural induction, the capability to develop in vitro into different neuronal subpopulations (cholinergic, dopaminergic, noradrenergic, somatostatinergic and some other peptidergic subpopulations without ongoing influences from the chordamesoderm (Duprat et al., 1987). Using the same in vitro model system, the aim of the present work was to delineate the abilities of these neuroblasts to develop GABAergic traits. The initial appearance and development of GABAergic phenotype has been quantitated by assaying the activity of glutamic acid decarboxylase (GAD). GAD activity was undetectable at the early gastrula stage (stage 8a) and was slightly measurable at the early neurula stage (stage 14- onset of the culture). It increased subsequently over the next 14 days in vitro. The temporal pattern of appearance and development of GAD activity in culture was in agreement with that observed in vivo. Immunocytochemical studies showed that GABA-like immunoreactivity was expressed in vitro in a subpopulation of neurons. Thus the developmental program for GAD expression and GABA phenotype maturation is acquired at least in some neuronal precursors. These data together with previously reported results on the expression of cholinergic, catecholaminergic and peptidergic phenotypes demonstrate that different neuronal subpopulations emerge near the end of gastrulation i.e. immediately after neural induction. The embryonic origin of this neuroblast heterogeneity remains to be determined.
Assuntos
Sistema Nervoso/embriologia , Neurônios/fisiologia , Pleurodeles/embriologia , Salamandridae/embriologia , Ácido gama-Aminobutírico/fisiologia , Animais , Células Cultivadas , Imunofluorescência , Gástrula/fisiologia , Glutamato Descarboxilase/metabolismo , Imuno-Histoquímica , Cinética , Sistema Nervoso/citologia , Fatores de Tempo , Ácido gama-Aminobutírico/análiseRESUMO
Vitamin C (Vit C) is an important antioxidant, exerts powerful neuroprotective brain effects and plays a role in neuronal development and maturation. Vit C is present in brain tissue at higher concentrations than in other organs, but its detailed distribution in brain is unknown. Immunohistochemical detection of this vitamin has been performed by using a highly specific antibody against Vit C. The aim of the present work was to analyze the distribution of Vit C in children's brainstems during postnatal development, comparing two groups of ages: younger and older than one year of life. In general, the same areas showing neurons with Vit C in young cases are also immunostained at older ages. The distribution of neurons containing Vit C was broader in the brainstems of older children, suggesting that brainstem neurons maintain or even increase their ability to retain Vit C along the life span. Immunohistochemical labeling revealed only cell bodies containing this vitamin, and no immunoreactive fibers were observed. The distribution pattern of Vit C in children's brainstems suggests a possible role of Vit C in brain homeostatic regulation. In addition, the constant presence of Vit C in neurons of locus coeruleus supports the important role of Vit C in noradrenaline synthesis, which seemed to be maintained along postnatal development.
Assuntos
Ácido Ascórbico/metabolismo , Tronco Encefálico/crescimento & desenvolvimento , Tronco Encefálico/metabolismo , Antioxidantes/metabolismo , Criança , Pré-Escolar , Feminino , Humanos , Imuno-Histoquímica , Lactente , Recém-Nascido , Locus Cerúleo/metabolismo , Masculino , Fibras Nervosas/metabolismo , Neurônios/metabolismoRESUMO
The aim of this study was to identify, at the ultrastructural level, the neuronal targets of dopamine afferents to the medial prefrontal and the anterior cingulate cortex of the adult rat. Since, in addition to pyramidal neurons, the cortical neuronal population mainly consists of GABAergic nonpyramidal intrinsic neurons, the simultaneous visualization of both dopamine- and GABA-containing neurons should leave the pyramidal neurons as the only unlabelled dopamine postsynaptic target. In this context, we used a double labelling immunocytochemical procedure: a pre-embedding PAP immunostaining to visualize monoclonal conjugated-dopamine (DA) antibody, followed by postembedding immunogold staining with a polyclonal conjugated-GABA antibody. In a single section sampling of 369 DA-immunoreactive (DA-IR) varicosities observed and the GABA-containing elements, 75% of the DA-IR terminals showed no indication of any contact with a GABA neuron. Twenty-five per cent were found in nonsynaptic contiguity with a GABA-immunoreactive neuronal element: axon, dendrite or cell body. When a DA varicosity was in nonsynaptic contiguity with a neuronal perikaryon (5% of cases), this cell was GABA positive. Ten per cent of the DA varicosities were contiguous to a GABA axon, but axoaxonic synapses in either direction were never observed. A symmetrical synapse between a DA varicosity and a GABA-containing dendrite was observed only once. The other 13 DA-IR terminals exhibiting a clear synaptic junction were apposed to nonGABA-containing dendrites, spines and shafts. Triads were observed in which a DA varicosity, forming or not a symmetrical synapse, was apposed to an unlabelled dendrite already receiving a symmetrical junction from another unlabelled axon. These data confirm and extend previous results designating the pyramidal cell dendritic tree as the main synaptic target of DA cortical afferents in rat and primate cerebral cortex. However, a direct effect of dopamine on a subpopulation of intrinsic GABA neurons cannot be excluded.
RESUMO
A previous report has shown that small diameter serotoninergic (5-HT) axons innervating the forebrain are selectively eliminated by treatment with an amphetamine derivative, (+/-)p-chloroamphetamine (PCA; Mamounas et al., [1991] J. Comp. Neurol. 314:558-586). It is well known that the spinal cord is the target of numerous monoaminergic fibers of different types. We have previously shown that the dorsal and ventral horns and the intermediolateral cell column are innervated by numerous serotonin-, noradrenaline- and dopamine-containing axons, including both thin axons with small varicosities and beaded axons with large varicosities. In all these regions, the large majority of fine indolaminergic fibers do not establish synaptic contacts, contrasting with the beaded axons which mostly exhibit synapses. The present work was conducted to study the effect of PCA on the monoaminergic innervation of the adult rat spinal cord. Animals received two subcutaneous doses of PCA 24 hours apart and were perfused 3 weeks later. Immunocytochemistry was performed to detect 5-HT, noradrenergic and dopaminergic fibers by using light and electron microscopy. Light microscopy revealed that PCA treatment caused a marked and selective elimination of the fine 5-HT-immunoreactive fibers, mainly found in the dorsal horn, but spared all other larger axons. This selective effect on the dorsal horn innervating thin 5-HT fibers was confirmed with the electron microscope by calculating the synaptic incidence(s) of monoaminergic innervation. These results suggest that fine and beaded 5-HT axons correspond to two anatomically, biochemically and pharmacologically different types of fibers, which could arise from two subpopulations of brainstem neurons. In addition, this drug could be used to provide an experimental animal, devoid of 5-HT nonsynaptic fibers, thereby facilitating a study on the role of dorsal horn nonsynaptic system in pain modulation.
Assuntos
Dopamina/metabolismo , Norepinefrina/metabolismo , Serotonina/metabolismo , Medula Espinal/efeitos dos fármacos , Medula Espinal/metabolismo , p-Cloroanfetamina/farmacologia , Animais , Comportamento Animal/efeitos dos fármacos , Imuno-Histoquímica , Masculino , Microscopia Eletrônica , Ratos , Ratos Sprague-DawleyRESUMO
The distribution of dopamine and dopamine-immunoreactive neurones was studied in the central nervous system of the snail Lymnaea stagnalis. The results from immunocytochemical labelling were compared with those from the application of the glyoxylic acid fluorescence method and 6-hydroxydopamine-induced pigment labelling. Comparisons were also made between the number of dopamine immunoreactive neurones and the dopamine content of the ganglia, measured by high-performance liquid chromatography. Dopamine immunocytochemistry proved to be superior to the other two histochemical techniques in terms of specificity and sensitivity. The 6-hydroxydopamine-induced pigment labelling failed to prove a useful tool for the in vivo identification of all dopamine-containing neurones. The distribution and number of dopamine-immunoreactive neurones and levels of biochemically measured dopamine in specific ganglia showed a close correspondence. By using the results of the dopamine immunocytochemistry and glyoxylic acid technique, a detailed map of dopamine-containing neurones was constructed. Dopamine-containing inter- and intra-ganglionic axon tracts were also demonstrated. The mapping of dopamine-containing neurones will facilitate further neurophysiological analysis of dopaminergic neural mechanisms in Lymnaea.
Assuntos
Sistema Nervoso Central/fisiologia , Dopamina/fisiologia , Lymnaea/fisiologia , Neurônios/fisiologia , Animais , Encéfalo/citologia , Encéfalo/crescimento & desenvolvimento , Sistema Nervoso Central/citologia , Bochecha/inervação , Dopamina/imunologia , Dopamina/metabolismo , Imunofluorescência , Gânglios/citologia , Gânglios/metabolismo , Glioxilatos/metabolismo , Hidroxidopaminas , Imuno-Histoquímica , Neurônios/metabolismo , Oxidopamina , Pleura/citologia , Pleura/crescimento & desenvolvimento , Coloração e RotulagemRESUMO
The morphology of the Dorsal Unpaired Median (DUM) neurones in the Terminal Abdominal Ganglion (TAG) of the adult male cockroach Periplaneta americana were described based on wholemount preparations and paraffin sections and by using anterograde and retrograde cobalt mapping, octopamine-like immunohistochemistry, and double immunofluorescence technique with both conjugated gamma-aminobutyric acid (GABA) and octopamine antisera. Among 60 +/- 6 neurones with large somata (diameter 40 to 60 microns) on the dorsal midline surface of the TAG that were stained with toluidine blue, about 36 efferent DUM neurones exhibited octopamine-like immunoreactivity. The DUM neurones were arranged in three clusters (anterior, median and posterior) corresponding to the 7th-11th abdominal ganglia of the fused TAG. Anterior efferent DUM neurones with one, two, and four pairs of lateral neurites entered segmental nerves VIIB; VIIB and phallic nerves; IXB and phallic nerves; VIIIA, IXA, X, and IX, respectively. Three octopamine-like immunoreactive DUM neurones innervating heart chambers via segmental nerves (VIIA, VIIIA, and IXA) in the last abdominal segments occurred within abdominal ganglia 7, 8, and 9. Together with octopamine-like immunoreactive efferent DUM neurones, GABA-like immunoreactive dorsal midline neurones with small somata (10 to 20 microns) also occurred within the median group. The spatial distribution of DUM neurones in the TAG suggested that they had their origins in the median neuroblast, as for DUM neurones in the grasshopper.
Assuntos
Abdome/inervação , Gânglios dos Invertebrados/anatomia & histologia , Gânglios dos Invertebrados/fisiologia , Neurônios/fisiologia , Periplaneta/anatomia & histologia , Periplaneta/fisiologia , Animais , Imunofluorescência , Gânglios dos Invertebrados/citologia , Sistema de Condução Cardíaco/metabolismo , Imuno-Histoquímica , Masculino , Terminações Nervosas/anatomia & histologia , Terminações Nervosas/fisiologia , Neurônios/ultraestrutura , Octopamina/metabolismo , Coloração e Rotulagem , Ácido gama-Aminobutírico/metabolismoRESUMO
The distribution of catecholaminergic amacrine cells has been investigated in rats by means of immunohistochemical labelling of wholemounted retinas. Two groups of catecholamine-containing cells could be distinguished on the basis of their catecholamine and biosynthetic enzyme content. Both groups could be stained with an anti-tyrosine hydroxylase (TH) antiserum. The first group was composed of large, strongly TH-immunoreactive stellate amacrine cells, located principally in the innermost row of the inner nuclear layer (INL) and sending processes to the outermost sublamina of the inner plexiform layer (IPL). Some were displaced in the IPL or in the ganglion cell layer (GCL). This first group of cells can be regarded as dopaminergic since they were also stained by an anti-dopamine (DA) antiserum. The second group was composed of small, weakly TH-positive cell bodies, located slightly more sclerad within the INL. Their processes were usually not labelled with anti-TH. Identical cells could be better visualized with an anti-phenylethanolamine-N-methyltransferase (PNMT) antiserum. Their processes were observed in the middle sublamina of the IPL. A great number of these cells were displaced in the GCL. They could be regarded as epinephrine cells. Concerning the density and distribution throughout the retina a striking difference was observed between the superior and inferior halves of the retina, whereas a lower difference was observed between the nasal and temporal regions. Almost all the PNMT-immunoreactive cells were located throughout the upper retina, whereas the DA-cells were especially concentrated in the upper temporal quadrant. The distribution of the DA cells parallels that of the ganglion cells whose density is also maximal in the upper temporal retina.
Assuntos
Catecolaminas/análise , Neurônios/classificação , Retina/citologia , Animais , Reações Antígeno-Anticorpo , Dopamina/análise , Técnicas Imunoenzimáticas , Masculino , Matemática , Norepinefrina/análise , Feniletanolamina N-Metiltransferase/análise , Ratos , Ratos Endogâmicos , Retina/enzimologia , Tirosina 3-Mono-Oxigenase/análiseRESUMO
We have investigated with light and electron microscope immunocytochemistry the aminergic and peptidergic innervation of Onuf's nucleus in adult baboons. This nucleus, located in the ventrolateral part of the sacral spinal cord (S2 and S3), is considered to control urethral and anal sphincters and penile muscles. By comparison of intact and transected spinal cords, we have found that serotoninergic innervation has two origins: first, supraspinal, innervating the whole nucleus, with a possible predominance in the dorsal half; and second, intraspinal, corresponding to the ventral half of the nucleus. Thyrotropin-releasing hormone innervation appears largely coincident with serotonin, both in intact and transected spinal cords. Noradrenaline is exclusively of supraspinal origin, as attested by its disappearance below the level of the section. Substance P, calcitonin gene-related peptide, and Leu- and Met-enkephalin, which profusely innervate Onuf's nucleus, are on the contrary not affected by the transection. They most likely originate from the cord itself or the dorsal root ganglia. Thus, Onuf's nucleus innervation in the baboon arises both from supraspinal and intraspinal sources. The present study provides an anatomical basis for both voluntary and reflex controls of excretory and sexual functions in a primate. The same neurotransmitter (serotonin) according to its cell origin and discrete topography could exert different influences upon the same effector system.
Assuntos
Neuropeptídeos/fisiologia , Norepinefrina/fisiologia , Papio/fisiologia , Serotonina/fisiologia , Medula Espinal/fisiologia , Animais , Peptídeo Relacionado com Gene de Calcitonina/fisiologia , Cordotomia , Encefalina Leucina/fisiologia , Encefalina Metionina/fisiologia , Imuno-Histoquímica , Masculino , Microscopia Eletrônica , Neurônios Aferentes/fisiologia , Medula Espinal/citologia , Substância P/fisiologia , Hormônio Liberador de Tireotropina/fisiologia , Micção/fisiologiaRESUMO
Anti-phosphatidylserine antibodies were raised in rabbits immunized with phosphatidylserine-polyacrylamide gels and with phosphatidylserine-cytochrome c vesicles. A solid-phase immunoenzymatic assay (ELISA) was developed to study the immune serum. The optimal conditions were defined and the technique used to obtain quantitative results. The anti-phosphatidylserine serum diluted to 1/1000 was tested against several phospholipids and was found to be highly specific to phosphatidylserine. This represents the first experimental demonstration of the specificity of antibodies raised against phosphatidylserine.