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Calretinin (CR) as a buffer and sensor protein plays an important role in regulatory processes of Ca²⁺ and anty-apoptotic cellular protection. In the present study, immunohistochemical peroxidase- antiperoxidase (PAP) method was used in order to determine the numbers, morphology, morphometry and distribution pattern of CR in neurons of the chinchilla’s claustrum (Cl) and endopiriform nucleus (EN). In Cl and EN the presence of several classes of neurons with different immunoreactivity to CR was found. In Cl, CR-immunoreactive (IR) neurons were predominantly found in close vicinity to insular border while CR-IR neurons were evenly scattered throughout EN. In general, immunoreaction to CR was observed in neuronal cytoplasm, nucleus and in fibres-like nerve extensions. Statistical analysis showed the differences between average large diameter as well as cross-sectional area of CR-IR neurons present in Cl and EN. It is suggested, that CR acting as a calcium binding protein may play a role in neuronal network. Further co-localization studies are necessary to fully elucidate neurophysiology and neuropathology of the chinchilla’s Cl and EN neurons.
Pituitary adenylate cyclase activating polypeptide (PACAP) is a neuropeptide existing in two variant forms (of either 27 or 38 residues), widely present in numerous organs and evoking multiple effects both in the central and peripheral nervous systems. The present study was undertaken to evaluate the distribution pattern of PACAP-27 expression in the ovine pancreas. Using double immunohistochemical stainings co-localizations of PACAP-27 with galanin, SP or CRF were studied in intrapancreatic neurons. In intrapancreatic ganglia, immunoreactivty to PACAP-27 was found in 87.6 ± 5.4% of PGP 9.5-positive intrapancreatic neurons but not in intraganglionic nerve fibres. Numerous PACAP-27-im-munoreactive nerve terminals were also observed between pancreatic acini and around small arterioles. No immunoreactivity to PACAP-27 was found in the endocrine pancreas. In 42.9 ± 6.2% of PACAP-27-immunoreactive intrapancreatic neurons the expression of galanin was also found. Statistically lower subpopulation (12.4 ± 4.0%) of intrapancreatic neurons exhibited simultaneously the immunoreactivity to PACAP-27 and SP. The expression of CRF was detected in the relatively smallest group (3.2 ± 1.4%) of PACAP-27-positive intrapancreatic neurons. The present results suggest that in the ovine pancreas PACAP-27 may play an important role as mediator of pancreatic functions. In PACAP-related pancreatic activities, a modulatory role of galanin, SP and to a lower extend of CRF is also likely.
Cocaine- and amphetamine-regulated transcript (CART) is a recently discovered neuropeptide thought to mainly act in most laboratory mammals and humans as anorexigenic factor. The expression of CART in wild living animals is barely known. In the present study immunohistochemical stainings were applied to identify CART-immunoreactive (IR) structures in the pancreas of European bison. Antibodies against neuronal marker Hu C/D were used to visualize intrapancreatic neurons. The expression of CART was detected in approx. 75% of Hu C/D-IR intrapancreatic neurons which may thus also act as interneurons. Additionally, in most intrapancreatic ganglia single CART-IR non-varicose nerve fibers running between neurons were found. Pancreatic blood vessels as well as intralobular ducts were sparsely innervated with CART-IR nerve fibers. Moderately numerous CART-IR nerve terminals were found to innervate the pancreatic endocrine and exocrine tissue. None of islet endocrine cells showed the expression of CART. No presence of CART-IR neuronal elements were found in external connective tissue capsule and septa penetrating inside to the organ. Our study is the first to outline the presence of some differences in CART-ergic innervation pattern of the pancreas between domestic and wild mammals. The lack of CART-IR endocrine islet cells in the pancreas of European bison is an interesting finding, nevertheless its significance is largely unknown at the moment and needs to be further investigated.
αCaMKII, widely occurring in the central nervous system, plays a significant role in cognitive processes. It is well known that diabetes is a risk factor that may trigger brain atrophy, cognitive dysfunction and finally lead to memory loss. Antioxidants richly present in bilberry fruits are believed to have significant effects on diabetes-related brain dysfunctions mainly due to their abilities to modulate neurotransmitter release that lead to reduction of the negative impact of free radicals on cognitive processes. The aim of the present research was to immunohistochemically investigate the expression patterns of αCaMKII in hippocampal neurons from non-diabetic, diabetic and diabetic rats fed with an extract of bilberry fruit. The obtained results show that in comparison to the control group, in diabetic rats hippocampal neurons immunoreactive (ir) to αCaMKII were swollen and the lengths of the neuronal fibres were reduced. Further study shows that in diabetic rats fed with bilberry fruit, αCaMKII-positive nerve fibres were significantly longer when compared to the groups of diabetic and control rats. Additionally, we observed statistically significant changes in the average larger diameter of αCaMKII-ir hippocampal neurons between groups of diabetic rats (with vs. Without supplement of bilberry fruit). The results of the present work suggest that antioxidants present in bilberry fruits influence the morphology of and possibly exhibit beneficial and neuroprotective effects on hippocampal neurons during diabetes. It is likely that changes in the appearance of αCaMKII-expressed hippocampal neurons may reflect the diabetes-evoked rise in Ca²⁺ level in the cerebral nerve terminals. The present research extends our knowledge of preventive mechanisms for cognitive dysfunctions occurring in the brain during diabetes.
Neuroendocrine cells (NEC) are a cell population in the gastrointestinal tract that plays a role in the regulation of the digestion process, satiety and nutrient homeostasis. NE cells express a variety of bioactive hormones that can undergo changes in response to different luminal stimuli, including multiple components, which are present in the diet. In recent years, a modern (hybrid) type of rye grain has been introduced to feed industry. The goal of the present study was to determine immunohistochemically whether the feeding of the pigs with population and hybrid rye grains may evoke adverse changes in the small and large intestines in terms of the expression of serotonin, glucagon-like peptide 1 (GLP1) and somatostatin. Feeding animals with population and hybrid rye grains resulted in a slight increase in serotonin-positive NE cells in the small intestine (but not in the large intestine). After feeding animals with population rye (but not with hybrid rye) grains, there was a decrease in the small intestine GLP1-immunoreactive NE cells was found. No changes in the expression of GLP1 were found in the large intestine of experimental animals. The numbers of somatostatin-IR NEC in the small and large intestines were not affected by feeding with either population or hybrid rye grains. In conclusion, we found that feeding pigs with hybrid and population rye grains started adaptive changes in NEC. However, those changes were not profound, which allows us to speculate that adverse effects of these rye grains have a minor (if any) impact on the gut hormone balance (and indirectly on the health status) of animals.
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