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Intracerebral haemorrhage is a devastating neurological disease with high mortality rate and poor prognosis. The most prominent manifestation of the disease are the movement disorders, but many patients also suffer from cognitive impairment. Taking into account vulnerability of the neurons located within the hilus of the dentate gyrus (HDG) to many brain insults we decided to study the effect of experimentally induced intracerebral haematoma on density of neurons expressing NogoA protein in HDG. In addition, we studied how administration of valproic acid and minocycline, the two drugs generally believed to be neuroprotective agents, influences the density of these neurons. Our study revealed that 4 weeks after intracerebral haematoma induction, minocycline and valproic acid treatment increased the densities of NogoA-ir neurons in the hilus of contralateral dentate gyrus once the data were compared to ipsilateral hemispheres within the same group. The analysis of contralateral hemisphere data, however, revealed increased densities of NogoA-positive neurons in haematoma and valproic acid treated animals when compared to contralateral hemispheres of control animals. The administration of minocycline was, however, able to alleviate this increase. These changes may influence the haematoma-induced reorganisation of neuronal circuitries in the dentate gyrus. (Folia Morphol 2014; 73, 3: 279–285)
The present study investigates the development of microglial and astroglial cells in the postnatal rat striatum, using immunohistochemical methods with panel antibodies that recognize macrophage antigens of unknown function - ED l, complement type 3 receptor- OX-42 (for microglia) and glial fibrillary acidic protein (for astrocytes). On the day of birth, EDl/OX-42- immunoreactive microglial cells present in the striatum represent ameboid microglia. Between PO and P10 we could observe the migration of ameboid microglial cells from neuroepithelial ventricular zone through internal and external capsules into the striatum. During the second postnatal week (PIO, P14) a considerable decline of ameboid EDl-immunoreactive microglial cells and an increase of the number of OX-42 positive ramified cells were observed. At P21 only OX-42 positive ramified cells were observed in the whole striatum. On the day of birth, only a few GFAP positive cells resembling radial glia were observed in the striatium. During the first postnatal week, the number of GFAP-positive cells increased significantly; they showed typical morphology of the astrocytes present in the adult animals. After P22 the final striatal population of astroglia was formed.
Intracerebral hemorrhage (IH) is a devastating form of cerebrovascular disease. Several studies points at glial response as one of the key factors in processes related to brain damage and repair after IH. The aim of the present study was to evaluate the influence of administration of minocycline and valproic acid on glial response after IH. 56 adult male Wistar rats (290 - 410 g) were used for the study. In 48 rats IH was induced by injection of 200 µl of autologous blood into the temporal lobe structures. Next animals were divided into three groups that received either minocycline (3 × 200 mg/kg), or valproic acid (2 × 45 mg/kg) or 0.09% saline (2 ml) for 7 consecutive days. 8 sham-operated animals served as controls. Animals from each group were successively sacrificed at 2, 4, 24 and 48 weeks after the hematoma induction. The intensity of glial response was based on qualitative and semiquantitative evaluations of immunostained sections. We observed that during the course of IH glial cells exhibit time-dependent changes in morphology and intensity of staining. 2 and 4 weeks after IH induction activated forms of astro- and microglia were observed near the border of hematoma as well as neighboring structures, while 24 and 48 weeks later they were present mainly around the glial scar and in the degenerating white matter. White matter structures also contained NOGO-A-immunoreactive oligodendrocytes. Administration of minocycline or valproic acid decreased the number of activated astro- and microglial cells in the white matter of hemisphere contralateral to site of injury at all time points. The number of oligodendrocytes was influenced only by minocycline. The obtained results indicate that minocycline and valproic acid administration modifies the number of glial cells in the white matter of hemisphere contralateral to the site injury. The study was financially supported by Polish Ministry of Science and Higher Education grant nr 3419/B/P01/2008/35.
It is postulated, that brain-derived neurotrophic factor (BDNF) have been implicated in the neurobiological mechanisms underlying brain plasticity after chronic stress. The objective of this study was to evaluate infl uence of chronic stress on brain plasticity measured by BDNF immunoreactivity in brain structures of young (P28) and adult (P360) rats. 26 male Wistar rats were exposed to 15 min daily open fi eld (OF) or forced swim test (FS) during three weeks. Fluorescent immunohistochemistry was used to localize BDNF positive cells in hypothalamic areas connected with stress response: both parvo- and magnocellular divisions of the paraventricular nucleus (PVp and PVm) and the supraoptic nucleus (SO). In animals aged P28 chronic OF i FS stress caused a statistically signifi cant (P<0.001) decline in the number of BDNF-ir cells in both parts of the PV and SO. In contrast, in rats P360 was not observed any change in the number of BDNF-ir cells after chronic OF stimulation compared to control in PVp and SO. In summary: age of rats subjected to chronic stimulation OF FS or stress had an impact on changes in the number of BDNF-ir cells in the tested hypothalamic nuclei.
Immunohistochemical study of the cholinergic innervation of the hippocampal calretinin-containing cells was conducted on 28 rat brains of postnatal ages: P0, P4, P7, P14, P21, P30 and P60. Sections with double immunostaining for vesicular acetylcholine transporter (VAChT; the marker of cholinergic cells, fibres and terminals) and calretinin were analysed using confocal laser-scanning microscope. Obtained data demonstrate that during development as well as in adult species calretinin-containing neurones in the rat hippocampus form sparse synaptic contact with VAChT-ir terminals. It seems probable that cholinergic innervation is not crucial for the functioning of CR-ir cells — probably they remain under the greater influence of a system other than the cholinergic system.
Fluoride alters the expression and post-translational modifications of extracellular matrix proteins in dentin. The aim of our study was to determine the effects of fluoride on type I collagen expression during the early stages of tooth germ development in rats. Pregnant dams were divided into three groups and fed a standard diet. From the fifth day of pregnancy the three groups received tap water with, respectively, trace amounts of fluoride (C), a low fluoride concentration (FL) or and a high fluoride concentration (FH). Changes in type I collagen expression and distribution were evaluated. The expression of type I collagen was restricted to the extracellular spaces of cells of mesenchymal origin. In the youngest animals the most intense immunoreactivity for type I collagen was detected in predentin of the FL group. Although the intensity of immunostaining increased in proportion to the age of the animals, the largest increase in the groups investigated was detected in the FL group. We concluded that a low concentration of fluoride can act as a stimulator of type I collagen deposition in the extracellular matrix of dentin, while high concentrations of fluoride have an opposite effect, acting as an inhibitor of type I collagen formation in dentin.
In the present study we wanted to check whether the expression of the c-Fos protein (the marker of cellular activity) appears in cells containing calcium-binding proteins (CaBPs) in animals exposed to the open field test. Eight adult Wistar rats were examined. In the first step the open field test was applied throughout 10 minutes. After perfusional fixation brains were frozen and cut on the cryostat in the coronal plane and stained with the standard immunohistochemical method. Sections were double stained for c-Fos and CaBPs: parvalbumin (PV), calbindin (CB), calretinin (CR). c-Fos positive cells were localized predominantly in layers II and III of the piriform cortex (PC). The double labeling study showed that neurons containing CaBPs are rarely c-Fos-immunoreactive. Often PV-positive and CB-positive fibers surround c-Fos-positive neurons in layers II and III in a form of a basket. It seems that cells containing CaBPs are not directly involved in the response to aversive stimuli but cells containing those calcium-binding proteins might influence directly c-Fos positive neurons of PC.
Immunohistochemical study of the cholinergic innervation of the hippocampal cells containing glutamic acid decarboxylase (GAD) and calcium binding proteins: parvalbumin (PV), calbindin D28k (CB) and calretinin (CR) was conducted on 5 adult rat brains. Analysis of sections with double immunostaining for vesicular acetylcholine transporter (VAChT; the marker of cholinergic cells, fibres and terminals) and respectively either GAD or PV, CB, CR, using confocal laser-scanning microscope shows that the intensive cholinergic innervations receive GAD, PV and CB-positive hippocampal cells. Cholinergic afferentations of the CR-positive neurones are considerably fewer.
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