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The aim of the study was the immunohistochemical evaluation of the impact of the age of animals treated with the sodium salt of glutamic acid on the behaviour of astrocytes of the infundibular nucleus (IN). Immunohistochemical peroxidase-antiperoxidase reactions were conducted on brain sections of 10-day-old (I) and 63-day-old (II) rats treated s.c with monosodium glutamate (MSG) in a dose of 4 g/kg b.w. for three consecutive days. The staining was performed using specific antibodies against glial fibrillary acidic protein (GFAP), S-100β protein and Ki-67 antigen. Cells immunopositive for the proteins under investigation were assessed morphologically and morphometrically in an Olympus BX51 light microscope with the Cell ^ D program. Statistically significant differences were tested using ANOVA and the non-parametric Kruskal-Wallis test. In the infundibular nucleus of 10-day-old rats treated with MSG, there was an increase in the number of GFAP, S-100β and Ki-67 immunopositive astrocytes without any changes in their morphology, which was typical of immature glia. In adult rats treated with MSG, a decrease in the number of cells expressing GFAP and S-100β was found. Most astrocytes had thick and weakly branched processes, in contrast to those observed in control animals. The results of our study showed a diverse behaviour of astroglia of IN in young and adult rats treated with MSG. In 10-day-old rats, hyperplasia of glia occurred, whereas in 63-day-old individuals there was a loss and hypertrophy of astrocytes, which may indicate a late stage of their reactivity. This information may contribute to targeting the therapy of diseases of the nervous system induced by the excitotoxic effects of glutamate.
The structure and functions of the oligodendrocytes in different areas of the brain in the normal process of aging in mammals is poorly known. The purpose of this study was to investigate three types of oligodendrocytes, their ultrastructure as well as morphology of myelin sheaths of nerve fibers in the central gray matter (substantia grisea centralis-SGC). The study was conducted on adult rats, 25-week-old and140-week-old rats. The animals were perfused with fixative through the left ventricle and the midbrain containing SGC were collected. Ultrathin sections were observed and photographed in an electron microscope. In both tested age groups oligodendrocytes were usually arranged in pairs. In adult rats, the SGC survey revealed a clear advantage and medium oligodenrocytów, and, rarely, dark cells with normal ultrastructure. In old rats, oligodendrocytes were dominated by medium and dark cytoplasm and less commonly with clear cells. The cytoplasm of the few bright and medium oligodendrocytes expressed the morphological changes manifested by the presence of varying electron densities and size of inclusions and the insulation of nerve fibers were also changed. The presence of the few bright oligodendrocytes in the SGC in old rats suggests that a few of their young forms of progenitor cells may arise, as in other areas of the brain normally aging in mammals. These new cells may participate in remielinization nerve fibers affecting the proper connections SGC with other brain areas.
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.
A thorough understanding of the mechanisms leading to the interaction between the sperm and the ovum in the process of fertilization in birds can facilitate more effective programming and control of the reproduction of these animals in breeding farms. In addition, it may allow the introduction of extracorporeal fertilization techniques, which may be important in the creation of transgenic animals and the reproduction of endangered species. In birds, the process of fertilization is not well known. It is conditioned by a series of interactions between mature reproductive cells. Oocytes are formed in the ovarian follicles of the left ovary. After ovulation, an ovum in the metaphase of the second meiotic division enters the oviduct along with the inner perivitelline layer (IPVL). It gets fertilized in this infundibulum. Male gametes are formed in paired testes located in the abdominal cavity. Sperm cells in the female reproductive tract do not require capacitation and are already fully capable of fertilization. As a result of internal insemination, male reproductive cells enter the oviduct. In this organ, they are selected and stored in the primary and secondary sperm storage tubules of the mucous membrane. They are released in batches shortly before ovulation. After reaching the oocyte, the sperm binds to the IPVL. This induces an acrosomal reaction that allows the male reproductive cells to penetrate to the surface of the oocyte, especially at the germinal pole. Next, as a result of physiological polyspermy, many sperm cells reach the ooplasm where they form haploid male pronucleus. This phenomenon is necessary to activate an polylecithal egg and produce a haploid female pronucleus. In the final stage, the female pronucleus merges with the single male pronucleus, which leads to the formation of a diploid zygote. The excess male pronuclei present in ooplasm are broken down by endonucleases (DNases). Understanding the mechanisms leading to the interaction between sperm and oocyte in birds may allow for more accurate programming and breeding of these animals in poultry farms and the introduction of extracorporeal fertilization techniques. In addition, it could be useful for the reproduction of endangered bird species.
Currently the structure and function of oligodendrocytes in the central nervous system (cns) in young individuals is known; however, there is no information about their morphology in the central gray matter (substantia grisea centralis-SGC) in old animals. The aim of this study was to investigate the morphology of oligodendrocytes in the semithin SGC sections of 6-month (5 pcs) and 3-year-old (5 pcs) male Wistar rats. The animals were anaesthetized with ketamine, perfused with fixative through the left ventricle, and the midbrains containing SGC were taken. Semithin sections were analyzed morphologically and photographs were made in light microscope Axiolab (Zeiss). In the semithin sections of the SGC of 6-month-old adult rats oligodendrocytes with mostly light and medium density cytoplasm were observed. Oligodendrocytes were located by different structures: blood vessels, neurons and astrocytes. Sometimes two cells with dark cytoplasm were side by side or near oligodendrocytes with a medium cytoplasm density. In old male rats oligodendrocytes with dark cytoplasm located at the capillary blood vessels, neurons and astrocytes were dominant in the SGC. Cells with light cytoplasm, which occurred mainly in blood vessels, were rarely encountered. The results indicate that in the SGC of 3-years-old individuals older forms with a dark cytoplasm are dominant, in contrast to 6-month-old rats which have younger oligodendrocytes with light cytoplasm.
The aim of this study was to track the immunoreactivity of parvalbumin and calbindin D28k buffering proteins in neurons of the dorsal raphe nucleus DRN of the adult male chinchilla. The reaction of peroxidase-antiperoxidase (PAP) using specific antibodies against these proteins was performed. In most neurons, with the exception of small oval and medium-sized, stellate neurons, weak immunostaining of parvalbumin was observed. On the contrary, very intense immunostaining for calbindin D28k was shown in almost all neurons of the examined area. The results indicate a slightly different distribution in the nucleus dorsal raphe in the adult male chinchilla than in the rat. This shows that mainly calbindin D28k is included in the regulation of intracellular calcium ion concentration in the nucleus dorsal raphe of chinchilla.
The aim of the study was to investigate acetylcholinesterase-immunoreactive neurons in the CA1 area of the hippocampus and in the striatum (CS) of rats receiving rebaudioside A (RebA) for 15 days. RebA is a steviol glycoside used in the production of sweeteners, and it has been shown that glycosides affect memory and learning processes. RebA was administrated to adult rats for 15 days at 1 mg of glycoside/ml of water (group I) and 2 mg of glycoside/ml of water (group II). An indirect immunohistochemical peroxidase-antiperoxidase reaction was performed on frontal slides containing the hippocampus and CS with the use of a monoclonal antibody against AChE. Neurons immunoreactive for the protein were assessed morphologically and morphometrically in hippocampal area CA1 and in the CS. Microscopical observations did not reveal significant morphological changes in immunopositive neurons, which suggests that the glycoside had no neurotoxic effect of these cells. Morphometric analyses did not show changes in the density of AChE-immunoreactive neurons. On the other hand, a decrease in reaction intensity was demonstrated in hippocampal area CA1 in group I and in the CS in both groups of animals receiving RebA. The results of our preliminary studies suggest that RebA affects cholinergic neurons.
The aim of the study was to determine the localization of estrogen receptors α in the parahippocampal gyrus (gyrus parahippocampalis) following 17β application in ovariectomized rabbits. Studies were carried out using an immunocytochemical method. Estrogen receptors a immunoreactivity in neurons and astrocytes were detected in the cellular layers of parasubiculum, perisubiculum and entorhinalis region. However ERα immunoreactivity was not detected in astrocytes ovariectomized rabbits in the experimental group where there was no E2 application. On the other hand, the neurons showed very weak colorization. The results obtained indicate that the structure of rabbit parahippocampal gyrus is under the influence of estrogen, which causes an increase in ERα expression.
The localisation of carletinin in the midbrains of 10 sexually mature chinchilla males was examined by carrying out the peroxidase-antiperoxidase immunocytochemical reaction using specific monoclonal antibody against calretinin. Intensive immunostaining for the protein was observed in the majority of fusiform, pyramidal, and stellate neurons of dorsal raphe nucleus. A similar calretinin distribution in neurons of this region to those observed in primates and rodents was demonstrated.
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