RESUMO
Mercury (Hg) is an environmental contaminant that poses great risk to human health. However, it is still widely used in artisanal gold-mining enterprises around the world, especially in developing countries. Methylmercury (MeHg) is produced environmentally by biomethylation of inorganic Hg present in water sediments, leading to its subsequent accumulation in the aquatic food chain. Due to its high metabolic rate, the Central Nervous System (CNS) is one of the main targets of MeHg. In the present study, we investigate the impact of chronic MeHg intoxication on NADPH diaphorase (NADPH-d) activity and astrocyte mobilization in the visual cortex of the rat. After 60 days of MeHg administration by oral gavage (0.04 mg/kg/day), tissue samples containing the visual cortex were submitted to measurements of Hg levels, NADPH-d activity, and GFAP immunohistochemistry for identification of astrocytes. MeHg intoxication was associated with increased Hg deposits and with reduced NADPH-d neuropil reactivity in the visual cortex. A morphometric analysis suggested that NADPH-d-positive neurons were mostly spared from MeHg harmful action and intoxicated animals had astrocytic activation similar to the control group. The decrease in NADPH-d neuropil reactivity may be due to the negative effect of chronic MeHg poisoning on both the synthesis and transport of this enzyme in afferent pathways to the visual cortex. The relative resistance of NADPH-d-reactive neurons to chronic MeHg intoxication may be associated with peculiarities in cell metabolism or to a protective role of nitric oxide, safeguarding those neurons from Hg deleterious effects.
Assuntos
Astrócitos/efeitos dos fármacos , Poluentes Ambientais/toxicidade , Compostos de Metilmercúrio/toxicidade , NADPH Desidrogenase/metabolismo , Neurônios/efeitos dos fármacos , Córtex Visual/efeitos dos fármacos , Animais , Astrócitos/enzimologia , Comportamento Animal/efeitos dos fármacos , Poluentes Ambientais/metabolismo , Ouro , Humanos , Masculino , Compostos de Metilmercúrio/metabolismo , Mineração , Neurônios/enzimologia , Ratos , Ratos Wistar , Córtex Visual/enzimologia , Córtex Visual/patologiaRESUMO
Ocular dominance plasticity (ODP) in the cat primary visual cortex (V1) is induced during waking by monocular deprivation (MD) and consolidated during subsequent sleep. The mechanisms underlying this process are incompletely understood. Extracellular signal-regulated kinase (ERK) is activated in V1 during sleep after MD, but it is unknown whether ERK activation during sleep is necessary for ODP consolidation. We investigated the role of ERK in sleep-dependent ODP consolidation by inhibiting the ERK-activating enzyme MEK in V1 (via U0126) during post-MD sleep. ODP consolidation was then measured with extracellular microelectrode recordings. Western blot analysis was used to confirm the efficacy of U0126 and to examine proteins downstream of ERK. U0126 abolished ODP consolidation and reduced both phosphorylation of eukaryotic initiation factor 4E (eIF4E) and levels of the synaptic marker PSD-95. Furthermore, interfering with ERK-mediated translation by inhibiting MAP kinase-interacting kinase 1 (Mnk1) with CGP57380 mimicked the effects of U0126. These results demonstrate that ODP consolidation requires sleep-dependent activation of the ERK-Mnk1 pathway.
Assuntos
Dominância Ocular/fisiologia , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Plasticidade Neuronal/fisiologia , Privação Sensorial/fisiologia , Sono/fisiologia , Córtex Visual/enzimologia , Potenciais de Ação/efeitos dos fármacos , Compostos de Anilina/farmacologia , Animais , Butadienos/farmacologia , Gatos , Dominância Ocular/efeitos dos fármacos , Fator de Iniciação 4E em Eucariotos/metabolismo , Feminino , MAP Quinase Quinase Quinases/antagonistas & inibidores , MAP Quinase Quinase Quinases/metabolismo , Masculino , Plasticidade Neuronal/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Neurônios/enzimologia , Nitrilas/farmacologia , Fosforilação/efeitos dos fármacos , Inibidores de Proteínas Quinases/farmacologia , Purinas/farmacologia , Sono/efeitos dos fármacos , Córtex Visual/efeitos dos fármacosRESUMO
To study the circuitry related to the ventral stream of visual information processing and its relation to the cytochrome oxidase (CytOx) modules in visual area V2, we injected anterograde and retrograde cholera toxin subunit B (CTb) tracer into nine sites in area V4 in five Cebus apella monkeys. The injection site locations ranged from 2° to 10° eccentricity in the lower visual field representation of V4. Alternate cortical sections, cut tangentially to the pial surface or in the coronal plane, were stained for CTb immunocytochemistry or for CytOx histochemistry or for Nissl. Our results indicate that the V4-projecting cells and terminal-like labeling were located in interstripes and thin CytOx-rich stripes and avoided the CytOx-rich thick stripes in V2. The feedforward projecting cell bodies in V2 were primarily located in the supragranular layers and sparsely located in the infragranular layers, whereas the feedback projections (i.e., the terminal-like labels) were located in the supra- and infragranular layers. V4 injections of CTb resulted in labeling of the thin stripes and interstripes of V2 and provided an efficient method of distinguishing the V2 modules that were related to the ventral stream from the CytOx-rich thick stripes, related to the dorsal stream. In V2, there was a significant heterogeneity in the distribution of projections: feedforward projections were located in CytOx-rich thin stripes and in the CytOx-poor interstripes, whereas the feedback projections were more abundant in the thin stripes than in the interstripes.
Assuntos
Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Retroalimentação , Rede Nervosa/enzimologia , Córtex Visual/anatomia & histologia , Córtex Visual/enzimologia , Vias Visuais/fisiologia , Animais , Cebus , Toxina da Cólera/metabolismo , Rede Nervosa/citologia , Neurônios/metabolismo , Campos VisuaisRESUMO
The cerebral cortex is often described as a composite of repeated units or columns, integrating the same basic circuit. The 'ice-cube' model of cortical organization, and 'canonical' circuit, born from insights into functional architecture, still require systematic comparative data. Here we probed the anatomy of an individual neuronal type within V1 to determine whether or not its dendritic trees are consistent with the 'ice-cube' model and theories of canonical circuits. In a previous report we studied the morphometric variability of NADPH-diaphorase (NADPH-d) neurons in the rat auditory, visual and somatosensory primary cortical areas. Our results suggested that the nitrergic cortical circuitry of primary sensory areas are differentially specialized, probably reflecting peculiarities of both habit and behavior of the species. In the present report we specifically quantified the dendritic trees of NADPH-d type I neurons as a function of eccentricity within V1. Individual neurons were reconstructed in 3D, and the size, branching and space-filling of their dendritic trees were correlated with their location within the visuotopic map. We found that NADPH-d neurons became progressively smaller and less branched with progression from the central visual representation to the intermediate and peripheral visual representation. This finding suggests that aspects of cortical circuitry may vary across the cortical mantle to a greater extent that envisaged as natural variation among columns in the 'ice-cube' model. The systematic variation in neuronal structure as a function of eccentricity warrants further investigation to probe the general applicability of columnar models of cortical organization and canonical circuits.
Assuntos
Dendritos/enzimologia , NADPH Desidrogenase/metabolismo , Córtex Visual/citologia , Vias Visuais/citologia , Animais , Mapeamento Encefálico , Análise por Conglomerados , Imageamento Tridimensional , Masculino , Células Piramidais/citologia , Células Piramidais/enzimologia , Roedores , Córtex Visual/enzimologia , Vias Visuais/fisiologiaRESUMO
Even though there is great regional variation in the distribution of inhibitory neurons in the mammalian isocortex, relatively little is known about their morphological differences across areal borders. To obtain a better understanding of particularities of inhibitory circuits in cortical areas that correspond to different sensory modalities, we investigated the morphometric differences of a subset of inhibitory neurons reactive to the enzyme nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) within the primary auditory (A1), somatosensory (S1), and visual (V1) areas of the rat. One hundred and twenty NADPH-d-reactive neurons from cortical layer IV (40 cells in each cortical area) were reconstructed using the Neurolucida system. We collected morphometric data on cell body area, dendritic field area, number of dendrites per branching order, total dendritic length, dendritic complexity (Sholl analysis), and fractal dimension. To characterize different cell groups based on morphology, we performed a cluster analysis based on the previously mentioned parameters and searched for correlations among these variables. Morphometric analysis of NADPH-d neurons allowed us to distinguish three groups of cells, corresponding to the three analyzed areas. S1 neurons have a higher morphological complexity than those found in both A1 and V1. The difference among these groups, based on cluster analysis, was mainly related to the size and complexity of dendritic branching. A principal component analysis (PCA) applied to the data showed that area of dendritic field and fractal dimension are the parameters mostly responsible for dataset variance among the three areas. Our results suggest that the nitrergic cortical circuitry of primary sensory areas of the rat is differentially specialized, probably reflecting peculiarities of both habit and behavior of the species.
Assuntos
Córtex Auditivo/citologia , Interneurônios/citologia , Interneurônios/enzimologia , NADPH Desidrogenase/metabolismo , Córtex Somatossensorial/citologia , Córtex Visual/citologia , Animais , Córtex Auditivo/enzimologia , Biomarcadores/metabolismo , Interneurônios/fisiologia , Masculino , Inibição Neural/fisiologia , Ratos , Ratos Wistar , Células Receptoras Sensoriais/citologia , Células Receptoras Sensoriais/enzimologia , Células Receptoras Sensoriais/fisiologia , Córtex Somatossensorial/enzimologia , Córtex Visual/enzimologiaRESUMO
Previous studies have shown a noticeable phenotypic diversity for pyramidal cells among cortical areas in the cerebral cortex. Both the extent and systematic nature of this variation suggests a correlation with particular aspects of cortical processing. Nevertheless, regional variations in the morphology of inhibitory cells have not been evaluated with the same detail. In the present study we performed a 3D morphometric analysis of 120 NADPH diaphorase (NADPH-d) type I neurons in the visual cortex of a South American Hystricomorph rodent, the diurnal agouti (Dasyprocta sp.). We found significant differences in morphology of NADPH-d type I neurons among visual cortical areas: cells became progressively larger and more branched from V1 to V2 and V3. Presumably, the specialized morphology of these cells is correlated with different sampling geometry and function. The data suggest that area-specific specializations of cortical inhibitory circuitry are also present in rodents.
Assuntos
NADPH Desidrogenase/metabolismo , Neurônios/citologia , Neurônios/metabolismo , Córtex Visual/citologia , Córtex Visual/metabolismo , Animais , Tamanho Celular , Análise por Conglomerados , Dendritos/enzimologia , Dendritos/metabolismo , Imageamento Tridimensional , Masculino , Neurônios/enzimologia , Fotomicrografia , Roedores , Córtex Visual/enzimologiaRESUMO
Tissue distribution of nitric oxide-synthases was investigated in the rat hippocampus and visual cortex under nutritional changes induced by modification of the litter size. Young (30-45-days-old) rats, suckled in litters formed by 3,6 or 12 pups (called small, medium and large litters, respectively), were studied by using nicotine-adenine-dinucleotide phosphate-diaphorase histochemistry (shortly, diaphorase), a simple and robust procedure to characterize tissue distribution of nitric oxide-synthases. We assessed morphometric features of the diaphorase-positive cells in visual cortex, and the neuropil histochemical activity in hippocampal CA1 and dentate gyrus using densitometry analysis. In the large-litter group, the labeled-cell density in white matter of area 17 was higher, as compared to the small-litter group. There was a clear trend, in the large-litter group, to lower values of soma area, dendritic field and branches per neuron, but the differences were not significant. Densitometry analysis of hippocampus revealed a significant increase in the relative neuropil histochemical activity of the dentate gyrus molecular layer in the larger litters, which may be associated to increased compensatory blood flow in the hippocampus. The pathophysiological mechanisms of the observed changes remain to be investigated.
Assuntos
Hipocampo/enzimologia , Lactação , Tamanho da Ninhada de Vivíparos , NADPH Desidrogenase/análise , Neurônios/enzimologia , Córtex Visual/enzimologia , Animais , Densitometria , Giro Denteado/enzimologia , Histocitoquímica , Neurópilo/enzimologia , Ratos , Ratos Wistar , DesmameRESUMO
The effects of methylmercury (MeHg) on histochemical demonstration of the NADPH-diaphorase (NADPH-d) activity in the striate cortex were studied in 4 adult cats. Two animals were used as control. The contaminated animals received 50 ml milk containing 0.42 µg MeHg and 100 g fish containing 0.03 µg MeHg daily for 2 months. The level of MeHg in area 17 of intoxicated animals was 3.2 µg/g wet weight brain tissue. Two cats were perfused 24 h after the last dose (group 1) and the other animals were perfused 6 months later (group 2). After microtomy, sections were processed for NADPHd histochemistry procedures using the malic enzyme method. Dendritic branch counts were performed from camera lucida drawings for control and intoxicated animals (N = 80). Average, standard deviation and Student t-test were calculated for each data group. The concentrations of mercury (Hg) in milk, fish and brain tissue were measured by acid digestion of samples, followed by reduction of total Hg in the digested sample to metallic Hg using stannous chloride followed by atomic fluorescence analysis. Only group 2 revealed a reduction of the neuropil enzyme activity and morphometric analysis showed a reduction in dendritic field area and in the number of distal dendrite branches of the NADPHd neurons in the white matter (P<0.05). These results suggest that NADPHd neurons in the white matter are more vulnerable to the long-term effects of MeHg than NADPHd neurons in the gray matter.
Assuntos
Gatos , Animais , Compostos de Metilmercúrio/intoxicação , NADPH Desidrogenase/metabolismo , Neurópilo/enzimologia , Córtex Visual/efeitos dos fármacos , Córtex Visual/enzimologia , Fluorescência , Mercúrio/análise , Microtomia , Neurônios/efeitos dos fármacos , Neurônios/patologia , Neurópilo/efeitos dos fármacos , Neurópilo/patologia , Córtex Visual/patologiaRESUMO
The effects of methylmercury (MeHg) on histochemical demonstration of the NADPH-diaphorase (NADPH-d) activity in the striate cortex were studied in 4 adult cats. Two animals were used as control. The contaminated animals received 50 ml milk containing 0.42 microgram MeHg and 100 g fish containing 0.03 microgram MeHg daily for 2 months. The level of MeHg in area 17 of intoxicated animals was 3.2 micrograms/g wet weight brain tissue. Two cats were perfused 24 h after the last dose (group 1) and the other animals were perfused 6 months later (group 2). After microtomy, sections were processed for NADPHd histochemistry procedures using the malic enzyme method. Dendritic branch counts were performed from camera lucida drawings for control and intoxicated animals (N = 80). Average, standard deviation and Student t-test were calculated for each data group. The concentrations of mercury (Hg) in milk, fish and brain tissue were measured by acid digestion of samples, followed by reduction of total Hg in the digested sample to metallic Hg using stannous chloride followed by atomic fluorescence analysis. Only group 2 revealed a reduction of the neuropil enzyme activity and morphometric analysis showed a reduction in dendritic field area and in the number of distal dendrite branches of the NADPHd neurons in the white matter (P < 0.05). These results suggest that NADPHd neurons in the white matter are more vulnerable to the long-term effects of MeHg than NADPHd neurons in the gray matter.
Assuntos
Compostos de Metilmercúrio/intoxicação , NADPH Desidrogenase/metabolismo , Neurópilo/enzimologia , Córtex Visual/efeitos dos fármacos , Córtex Visual/enzimologia , Animais , Gatos , Mercúrio/análise , Neurônios/efeitos dos fármacos , Neurônios/patologia , Neurópilo/efeitos dos fármacos , Neurópilo/patologia , Córtex Visual/patologiaRESUMO
We studied the tangential distribution of cytochrome c oxidase (CytOx)-rich blobs in four striate cortices of three normal monkeys (Macaca mulatta). The spatial density and cross-sectional area of blobs were analyzed in CytOx-reacted tangential sections of flat-mounted preparations of the striate cortex (V1). Well-delimited CytOx-rich blobs were found in the middle portion of cortical layer III of the V1. Throughout the binocular field representation, the spatial density of blobs was nearly constant with a mean value of four to five blobs per mm2. In the monocular portions of V1, however, blob spatial density diminished. In all cases, the mean cross-sectional area of blobs was constant in the V1. The small variation of CytOx blob topography with visual field eccentricity contrasts with the variation described in previously published material.