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The purpose of this study was to trace the immunoreactivity of the calcium binding protein calretinin in the periaqueductal gray matter of the midbrain of chinchillas. For this study the midbrains of five sexually mature male chinchillas were used. The immunoreactivity of this protein in this species has never been investigated up till now. The localization of its activity was examined by carrying out peroxidase-antiperoxidase (PAP) reaction using a mouse specific monoclonal antibody against calretinin. An intensive immunostaining for calretitin was observed in all the neurons in the dorsal and dorso-lateral periaqueductal gray matter. The results of the studies obtained suggest a similarity in the distribution of calretitin as seen in the neurons of periaqueductal gray matter of rats. This indicates that calretinin is involved in the regulation of intracellular calcium ion concentration. In this manner it can influence the proper functions of the neurons of the periaqueductal gray matter of the midbrain of the chinchilla.
The aim of this study was to investigate changes of calretinin immunoreactivity in neurons and neuropil of the dorsal raphe nucleus (DRN) after subcutaneous administration of monosodium glutamate (MSG) to adult rats. Studies were conducted on 60-day-old male rats. The animals were divided into a control group (C) and two other groups receiving MSG at a dose of 2g/kg b.w. (I) and 4g/kg b.w. (II) subcutaneously for 3 consecutive days. Immunohistochemical peroxydese-antiperoxydase reaction was conducted with the use of a specific anti-calretinin (CR) antibody on brain slides containing DRN of 63-day-old rats. The cells and neuropil were morphologically and morphometrically analysed under the light microscope Olympus BX51. Statistically significant differences were studied with ANOVA and nonparametric Kruskal-Wallis test. In 63-day-old rats, in DRN: dorsal (DRNd), ventral (DRNv) and interfascicular (DRNif) parts, in animals receiving MSG (I and II), there was a decrease in CR- immunoreactivity in neurons and neuropil in comparison to control rats. Only in the ventrolateral part (DRNvl) a few intensively stained CR-immunoreactive cells were demonstrated. Light microscope observations were confirmed by morphometric analyses. In the DRNd and DRNv of rats receiving MSG (I and II) a decrease in average CR-immunoreactive neuron density was shown in comparison to the C group. In the DRNvl part, a statistically significant decrease in the analysed parameter was present only in I group of animals. Conversely, in DRNif no statistically significant differences were shown between studied groups of rats. In the DRN of animals receiving MSG (I and II) a decrease in average digital immunostaining intensity for CR occurred in neurons and neuropil. The obtained results demonstrated a decrease in CR immunostaining intensity level in neurons and neuropil and a decrease in density of studied protein immunoreactive cells under the influence of subcutaneous administration of MSG to adult rats. These results suggest that MSG may cause neuronal death as a result of oxidative stress or it can alter a calretinin conformation in cells after binding to calcium ions.
The main aim of the study was to investigate the intracellular localization of the following calcium-binding proteins: parvalbumin, calbindin and calretinin. 15 sexually mature chinchilla males (about 1.5 years old) were used in the examination. The hippocampus was collected from each immediately after the slaughter, fixed and properly prepared for immunohistochemical examinations. Peroxidase-anti-peroxidase (PAP) reaction was carried out using specific antibodies against parvalbumin and calbindin D28k, as well as calretinin. Our own examination results have shown cytoplasmic as well as nuclear reactions in the examined regions of the hippocampal areas (CA1-CA4) and dentate gyrus. Only in the CA2 area was no nuclear reaction observed for the examined proteins, as well as in the CA1 area for calretinin. Intracellular localization of calcium-binding proteins proves that regulatory functions of parvalbumin, calbindin and calretinin lead to neuronal plasticity, i.e. to a change of their activity. Therefore, calcium-binding proteins may be indirectly involved in the regulation of metabolic processes affecting basic vital functions of neurons.
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