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Expression of substance-P in human neurons of trigeminal ganglia has been investigated by immunohistochemistry and morphometry. These neurons constituted 12.8% to 32.6% of the total neuronal population in the trigeminal ganglia. Substance-P positive granulations were concentrated around the nucleus, distributed focally in neuroplasm or dispersed over the neuroplasm. Morphometric analysis has indicated the presence of three populations of SP-positive cells: small, medium-sized and large. The results suggest a functional differentiation on the level of the first neurons of the afferent path of the stomatognatic system. Substance-P is likely to play a role in the transmission not only of nociceptive impulses but also of those involved in the mechano-functional stimulation of system activities.
The differences between pluripotent and differentiated cells include stage specific chromatin structure and transcriptional hierarchy which are both regulated by and orchestrated with the epigenetic events. Such events include alterations in DNA methylation, histone modifications, polycomb gene group and noncoding RNA expression. In this lecture the overview will be given on chromatin dynamics and epigenetic modification status during neural stem cell development. Examples of regulatory machineries responsible for gene repression at each stage of neural stem cell development will be indicated. Neural stem cells are characterized by their ability to give rise to multiple neural lineages, including neurons, astrocytes, and oligodendrocytes. Previously we have obtained neural stem cells from human cord blood (HUCB-NSC) which has been investigated by our group for their ability to be reprogrammed or differentiated using combination of small molecules as epigenetic modulators. It was demonstrated that the influence of small chemicals: histone deacetylases (Trichostatin A -TSA) and methyltransferases (RG-108) on the expression of Oct4, Sox2, Rex1 and Nanog genes depended on developmental stages of HUCB-NSC. Incubation for 5 days in reprogramming conditions followed by short time culture (3 days) in ESCM (Embryonic Stem Cell Medium) on Matrigel resulted with only partial stimulation of the investigated pluripotency markers. Nevertheless, the differences in expression pattern between tested treatment conditions were observed. Cells grown under Serum Free culture conditions treated with a combination of epigenetic inhibitors as well as recombinant proteins after longer incubation in ESCM on Matrigel were able to gain full iPS morphology and showed continuous expression of pluripotent genes. None of the mentioned above factors were alone sufficient to reprogram NSC to stable pluripotency state. Additionally the mechanism of regulation DNMTs and HDACs genes (namely DNMT 3B and HDAC1) by methyltransferases and histone deacetylases inhibitors and their role in reprogramming and differentiation process of HUCB-NSC have been tested. The present study demonstated that small molecules such as TSA and RG108 together with reprogramming proteins in lowered oxygen conditions can change epigenetic status of cells and activate and sustain pluripotent state in HUCB-NSC. In conclusion it is evident that the developmental stage of the cells and epigenetic modulation play an important role in the induction of pluripotency genes expression. Sponsored by grant from Polish Ministry of Scientific Research and Higher Education Nr 5978/B/PO1/2010/38
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In this study we investigated the potential neuroprotective effect of 2-oxoglutarate (2-OG) on the hippocampus in the transient vessel occlusion ischemia model in the Mongolian gerbil. The morphological and biochemical studies were performed at 7 days after occlusion of carotid arteries. The acute reduction of NeuN-positive neurons in the CA1 pyramidal layer of the hippocampus was accompanied by increased staining intensity for GFAP-positive astrocytes, indicative of glial reaction. The neuron death in the CA1 area coincided with a strong 2.4 fold decrease in the membrane forms of neuronal cell adhesion molecules and elevated levels of astrocyte-specific proteins (soluble GFAP to 2,6 times; filament GFAP to 1,5 times; calcium-binding protein S-100b to 1,6 times). Treatment with 2-oxoglutarate (2.28 g/l drinking water) for between 7 and 21 days attenuated the neuronal death and reactive astrogliosis in this model of experimental ischemia by 20-50%. Our results suggest that 2-OG may prevent the disturbances of neural cells that usually take place during ischemic pathology.
Because of their potential for self-renewal and the ability for generating many differentiated cell types, progenitor cells are a key player in regenerative and repair processes. In the central nervous system, pools of these cells have been identified in two regions: the subgranular zone of hippocampal gyrus dentatus and the subventricular zone. Neural stem cells that reside in these regions are subject to a specific neurogenesis-stimulating and -regulating environment called ‘niche’. Our model of surgical brain injury (SBI) opens the avenues for studying the mechanisms of repair and reconstruction of brain cortex and enables demonstrating the presence of possible vascular niches in the peri-lesion zone. The present studies were aimed at characterizing of the immune phenotype of the cells that populate this region. The peri-lesion area of the brain cortex showed the presence of dying neurons and glial cells since the first postlesion day. Simultaneously, activated microglial cells and astrocytes appeared, and part of the latter formed a scar on the surface of the damaged cortex. Another fractions of the cells that appeared following the SBI in both the lumen and the vicinity of blood vessels expressed either the macrophagal/monocytic marker CD14, or the marker of hematopoietic progenitor cells and small vessel endothelium CD34. Beginning on the first post-SBI day, the peri-lesion area showed also the presence and accumulation of a variety of cells with immature phenotypes. These included immature endothelial cells building new blood vessels (angiogenesis) and cells with phenotypes of other brain parenchyma-forming cell subpopulations: (1) nestin-positive astroglial and non-glial cells, (2) cells expressing the marker of juvenile astrocytes vimentin-positive, and (3) cells showing doublecortin immunoreactivity (the marker of early differentiated neurons). These results clearly indicate that during the early post-SBI period the peri-lesion zone is being populated by a heterogenic pool of morphologically immature cells that most likely herald the advent of reconstruction and/or repair of the injured brain region. Supported by the Polish Ministry of Science and Higher Education grant No N N404 522838
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