RESUMO
Glial activation and degeneration are important outcomes in the pathophysiology of acute brain and spinal cord injury (SCI). Our main goal was to investigate the pattern of glial activation and degeneration during secondary degeneration in both gray matter (GM) and white matter (WM) following SCI. Adult rats were deeply anesthetized and injected with 20 nmol of N-methyl-D-aspartate (NMDA) into the ventral horn of rat spinal cord (SC) on T7. Animals were perfused after survival times of 1, 3, and 7 days. Ten-micrometer sections were submitted to immunocytochemistry for activated macrophages/microglia, astrocytes, oligodendrocytes, and myelin. Astrocyte activation was more intense in the vacuolated white matter than in gray matter and was first noticed in this former region. Microglial activation was more intense in the gray matter and was clear by 24 h following NMDA injection. Both astrocytosis and microglial activation were more intense in the later survival times. Conspicuous WM vacuolation was present mainly at the 3-day survival time and decreased by 7 days after the primary damage. Quantitative analysis revealed an increase in the number of pyknotic bodies mainly at the 7-day survival time in both ventral and lateral white matter. These pyknotic bodies were frequently found inside white matter vacuoles like for degenerating oligodendrocytes. These results suggest a differential pattern of astrocytosis and microglia activation for white and gray matter following SCI. This phenomenon can be related to the different pathological outcomes for this two SC regions following acute injury.
Assuntos
Gliose/patologia , Microglia/metabolismo , Oligodendroglia/patologia , Traumatismos da Medula Espinal/patologia , Traumatismos da Medula Espinal/fisiopatologia , Animais , Imuno-Histoquímica , Ativação de Macrófagos/fisiologia , Masculino , Neurônios/patologia , Ratos , Medula Espinal/patologia , Medula Espinal/fisiopatologia , Fatores de TempoRESUMO
Nitric oxide (NO) has been implicated in neurotoxicity and cerebral blood flow changes in chronic neurodegeneration, but its activity in the mammalian prion diseases has not been studied in detail. Nicotine adenine dinucleotide phosphate (NADPH)-diaphorase (NADPH-d) histochemistry is a simple and robust histochemical procedure that allows localization of the tissue distribution of NO synthases. The aim of the present study is to assess whether NADPH-d histochemical activity is altered in the hippocampus in the ME7 model of prion disease in C57BL/6J mice. At early and late stages after the initiation of the disease we assessed features of the NADPH-d positive cells and the neuropil histochemical activity in CA1 and dentate gyrus using densitometric analysis. In C57BL/6J mice 13 weeks postinjection of the prion agent ME7, when behavioural changes first become apparent, neuropil NADPH-d histochemical staining increases, whereas at late stages it decreases dramatically. Both type I and type II NADPH-d positive cells were found to survive throughout the hippocampal formation into the late stages of the disease, but diaphorase activity was reduced in dendritic branches and abnormal varicosities were present in both dendritic and axonal processes of NADPH-d positive type I cells. The pathophysiological implications of the results remain to be investigated but both blood flow alteration and NO neurotoxicity may be features of the disease.
Assuntos
Hipocampo/patologia , NADPH Desidrogenase/metabolismo , Neurônios/patologia , Neurópilo/patologia , Doenças Priônicas/patologia , Algoritmos , Animais , Contagem de Células , Dendritos/metabolismo , Dendritos/patologia , Densitometria , Hipocampo/metabolismo , Imuno-Histoquímica , Camundongos , Camundongos Endogâmicos C57BL , Neurônios/metabolismo , Neurópilo/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase/biossíntese , Óxido Nítrico Sintase Tipo I , Doenças Priônicas/metabolismo , Fluxo Sanguíneo Regional/fisiologia , Sinapses/fisiologia , Fixação de TecidosRESUMO
We have estimated photoreceptor convergence to M and P retinal ganglion cells of two closely related nocturnal (owl monkey, Aotus) and diurnal (capuchin monkey, Cebus) anthropoids. Rod convergence is higher in the owl monkey retina while cone convergence to both M and P cells are very similar in the retinas of the owl monkey and the capuchin monkey. These results indicate that during evolution, the owl monkey retina has undergone changes compatible with a more nocturnal lifestyle, but kept a cone to ganglion cell relation similar to that found in diurnal primates.
Assuntos
Gânglios Sensitivos/anatomia & histologia , Células Fotorreceptoras de Vertebrados/citologia , Retina/anatomia & histologia , Animais , Aotus trivirgatus , Cebus , Contagem de Células , Feminino , Masculino , Vias Visuais/anatomia & histologiaRESUMO
Scrapie is a prion disease which occurs naturally in sheep and which can be transmitted experimentally to rodents. After intracerebral injection of ME7 into mouse, an atypical inflammatory response, characterized by T-lymphocytes and activated microglia is present early in the course of the disease. In the present work, we have investigated the relationship between this inflammatory response, astrocytosis and neuronal loss along the visual pathway after intraocular injection (intraocular) of ME7 in C57BL/6J mice. We have demonstrated that microglia activation and T-lymphocyte recruitment accompanies the spread of prion pathology along the visual pathway and in the early stages of the disease is restricted to the subcortical visual pathway. Inflammation was also present in non-visual areas in association with PrPsc deposition at late stages of the disease, possibily indicating that diffusion of the scrapie agent also contributes to the spread of the disease. After intraocular injection of the prion agent, the disease is believed to be transported into the brain via axons of retinal ganglion cells (RGCs). Despite the high levels of infectivity reported to be present in the retina early in the disease after intraocular injection of ME7, retinal pathology has not been extensively investigated. We have studied the RGCs response in whole mount retinas after intraocular injection of ME7. We have shown that RGCs degenerate after intraocular injection of ME7 whereas amacrine cells, retinal interneurones, are more resistant. Our results suggest that two distinct population of neurones, exposed in vivo at the same time to the same agent scrapie strain, show different susceptibility to the toxic effects of PrPsc.
Assuntos
Príons , Degeneração Retiniana/patologia , Células Ganglionares da Retina/patologia , Scrapie/patologia , Animais , Contagem de Células , Olho , Feminino , Injeções , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Retina/patologia , Análise de Sobrevida , Fatores de TempoRESUMO
Male Cebus monkeys are all dichromats, but about two thirds of the females are trichromats. M and P retinal ganglion cells were studied in the male Cebus monkey to investigate the relationship of their morphology to retinal eccentricity. Retinal ganglion cells were retrogradely labeled after optic nerve deposits of biocytin to reveal their entire dendritic tree. Cebus M and P ganglion cell morphology revealed by biocytin retrograde filling is similar to that described for macaque and human M and P ganglion cells obtained by in vitro intracellular injection of HRP and neurobiotin. We measured 264 and 441 M and P ganglion cells, respectively. M ganglion cells have larger dendritic field and cell body size than P ganglion cells at any comparable temporal or nasal eccentricity. Dendritic trees of both M and P ganglion cells are smaller in the nasal than in the temporal region at eccentricities greater than 5 mm and 2 mm for M and P ganglion cells, respectively. The depth of terminal dendrites allows identification of both inner and outer subclasses of M and P ganglion cells. The difference in dendritic tree size between inner and outer cells is small or absent. Comparison between Cebus and Macaca shows that M and P ganglion cells have similar sizes in the central retinal region. The results support the view that M and P pathways are similarly organized in diurnal dichromat and trichromat primates.
Assuntos
Percepção de Cores/fisiologia , Dendritos/fisiologia , Células Ganglionares da Retina/citologia , Animais , Cebus , Contagem de Células , Lisina/análogos & derivados , Masculino , Células Ganglionares da Retina/fisiologia , Visão Ocular/fisiologiaRESUMO
After partial transection of one optic nerve in adult cats the majority of beta retinal ganglion cells degenerate and die 1 week after axotomy, whilst other cell classes degenerate slowly and survive for a long period after the lesion. We have investigated the effects of intravitreal and intraperitoneal injections of MK-801, a NMDA-glutamate receptor antagonist, on the early degeneration of retinal ganglion cells after partial optic nerve section. Control animals received saline intravitreal injections. Retinal flat mounts were retrogradely labelled with horseradish peroxidase and counterstained with Cresyl Violet. We evaluated the ganglion cell loss in the three experimental groups 1 week after lesion and compared them with normal uninjured controls and injured untreated retinae. In untreated retinae 49% of ganglion cells die 1 week after the lesion. Systemic MK-801 or saline prolonged survival of 41% of retinal ganglion cells that would die without treatment. Intravitreal MK-801 or saline prolonged survival of 71% of retinal ganglion cells that would die without treatment, but the results of saline administration had a larger range of variability. In untreated retinae many pyknotic cells were observed. They decreased in number after systemic MK-801 treatment and in some retinae treated with intravitreal injections of saline solution. There were no pyknotic cells after local, intravitreal MK-801 treatment. These results support the hypothesis that NMDA-receptor mediated neurotoxicity plays an important role in the early retinal ganglion cell death after retrobulbar axotomy. They also support the existence of an endogenous source of neurotrophins whose release is triggered by eyeball injury. We conclude that the early death of beta retinal ganglion cells after axotomy occurs by a mechanism that can be controlled by neurotrophins and antagonists to NMDA-glutamate receptors.
Assuntos
Maleato de Dizocilpina/farmacologia , Traumatismos do Nervo Óptico , Receptores de N-Metil-D-Aspartato/antagonistas & inibidores , Células Ganglionares da Retina/efeitos dos fármacos , Animais , Gatos , Sobrevivência Celular/efeitos dos fármacos , Maleato de Dizocilpina/administração & dosagem , Feminino , Histocitoquímica , Peroxidase do Rábano Silvestre , Masculino , Degeneração Neural , Células Ganglionares da Retina/citologiaRESUMO
The topography of M ganglion cell distribution was studied in the retinae of two New World monkey species, the diurnal capuchin monkey Cebus apella and the nocturnal owl monkey Aotus azarae. Retinal whole mounts were stained by the neurofibrillar method of Gros-Schultze. As occurs with other diurnal primates, the Cebus M-ganglion cell density peaks in the foveal slope and declines towards the periphery. In the Aotus retina, the M ganglion cell density peaks in the area centralis and declines toward the periphery. In both species the cell density in the temporal, dorsal, and ventral meridians are similar for equivalent eccentricities. The cell density in the nasal meridian is higher than in the other meridians. The naso-temporal density ratio ranges between 1.2 and 4.3 in the Cebus and 1.6 and 2.2 in the Aotus. The total number of M-ganglion cells was 140,300 and 74,000 in the Cebus and Aotus retinae, respectively, corresponding to about 10% and 15.4% of the total retinal ganglion cell population in these species. The results indicate that M ganglion cells are similarly organized in both diurnal and nocturnal simians, but may be proportionally more important for the nocturnal species.
Assuntos
Aotus trivirgatus/anatomia & histologia , Cebus/anatomia & histologia , Ritmo Circadiano/fisiologia , Células Ganglionares da Retina/citologia , Animais , Aotus trivirgatus/fisiologia , Cebus/fisiologia , Contagem de Células , Tamanho Celular , Feminino , Masculino , Células Ganglionares da Retina/classificaçãoRESUMO
M and P retinal ganglion cell morphology revealed by biocytin retrograde labelling was compared in two closely related New-World monkeys, Cebus and Aotus, to investigate whether nocturnal and diurnal species of primates have similar cell classes. Monkey and cat ganglion cells from regions of matching cell class densities were also compared. Cat alpha, cat beta, Aotus M, and Cebus M cells were similar in many aspects, but Cebus M cells had higher branching density. Cebus and Aotus P cells formed a distinct group and represent a primate specialization common to diurnal and nocturnal simians.
Assuntos
Aotus trivirgatus/anatomia & histologia , Cebus/anatomia & histologia , Células Ganglionares da Retina/ultraestrutura , Animais , Aotus trivirgatus/metabolismo , Gatos/anatomia & histologia , Gatos/metabolismo , Cebus/metabolismo , Dendritos/metabolismo , Dendritos/ultraestrutura , Células Fotorreceptoras/metabolismo , Células Ganglionares da Retina/metabolismo , Especificidade da EspécieRESUMO
The early responses of cat retinal ganglion cells to axotomy have been examined using neurofibrillar and Nissl-stained wholemounts. We were interested to learn whether the enhanced neurofilament expression, seen in a number of neuronal systems, was also present in different neuronal populations of the cat retina and could be used to study the distribution of these cells. We found that beta ganglion cells degenerate very rapidly after axotomy with the nuclei becoming pyknotic within a few days. Few beta cells showed increased neurofibrillar staining of the dendrites. The cell body degenerated prior to any visible degenerative changes in the axon. A proportion of the alpha and gamma ganglion cells degenerated in the first two to three weeks after axotomy. The alpha cells underwent markedly enhanced neurofibrillar staining of their dendrites prior to degeneration. The Nissl material of the cell bodies diminished as the cells degenerated but we have not observed pyknotic nuclei. The dendritic trees of some axotomised gamma cells were also revealed by the neurofibrillar stain three weeks after axotomy. These results show that retinal ganglion cells do not degenerate by a dying back process. We suggest that the rapid degeneration of the beta ganglion cell population comes about by excitotoxic cell death, a consequence of their large glutamatergic input from bipolar cells. The degenerating beta ganglion cells have the morphological appearance of cells undergoing apoptosis.
Assuntos
Neurofibrilas/patologia , Traumatismos do Nervo Óptico , Células Ganglionares da Retina/patologia , Animais , Gatos , Feminino , Masculino , Coloração pela PrataRESUMO
The distribution of ganglion cells and displaced amacrine cells was determined in whole-mounted Aotus retinae. In contrast to diurnal simians, Aotus has only a rudimentary fovea. Ganglion cell density decreases towards the periphery at approximately the same rate along all meridians, but is 1.2-1.8 times higher in the nasal periphery when compared to temporal region at the same eccentricities. The total number of ganglion cells varied from 421,500 to 508,700. Ganglion cell density peaked at 15,000/mm2 at 0.25 mm dorsal to the fovea. The displaced amacrine cells have a shallow density gradient, their peak density in the central region is about 1500-2000/mm2 and their total number varied from 315,900 to 482,800. Comparison between ganglion cell density and areal cortical magnification factor for the primary visual cortex, area 17, shows that there is not a simple proportional representation of the ganglion cell distribution. There is an overrepresentation of the central 10 deg of the visual field in the visual cortex. The present results for Aotus and the results of a similar analysis of data from other primates indicate that the overrepresentation of the central visual field is a general feature of the visual system of primates.