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The fate choice of neural progenitors could be dictated by local cellular environment of adult CNS. The aim of our study was to investigate the effect of hippocampal tissue on the differentiation and maturation of the oligodendrocyte NG2 precursors. Methods: The hippocampal slice culture was established from the brains of 7-day old rats. The NG2 precursors, obtained from a 12-day old mixed primary culture of neonatal rat hemispheres, were labeled with CMFDA and seeded on hippocampal slices. After 7ñ14 days in co-culture, the cells were stained with neural markers. Results: The NG2 cells differentiated predominantly into oligodendrocytes, presenting various stages of maturation: progenitors NG2+, O4+ and fi nally mature Galc-positive cells. However, except for a few cells with astrocyte-specifi c S100β staining, a considerable number of these cells differentiated into TUJ+ and MAP-2+ neurons. Moreover, a certain population of these cells preserved proliferative properties of primary precursors, as revealed by the Ki67 expression. Conclusions: Neuronal microenvironment provided by the culture of hippocampal slices is potent to induce neurogenesis from oligodendrocyte NG2+/PDGFRα+/CNP+ progenitors and promotes their differentiation not only into macroglia but also into neurons. It also sustains their proliferative capacity. The results indicate a crucial role of the local cellular environment in fatedecision of primary NG2+ multipotent neural progenitors, which may affect their behavior after transplantation into CNS. Supported by MSHE grant N40101832 /0296
The effect of 2-aminobicyclo[2.2.1]heptan-2-carboxylic acid (BCH), an L-leucine nonmetabolizable analogue and an allosteric activator of glutamate dehydrogenase, on glucose and glutamine synthesis was studied in rabbit renal tubules incubated with alanine, aspartate or proline in the presence of glycerol and octanoate, i.e. under conditions of efficient glucose formation. With alanine+glycerol+octanoate the addition of BCH resulted in a stimulation of alanine and glycerol consumption, accompanied by an increased glucose, lactate and glutamine synthesis. In contrast, when alanine was substituted by either aspartate or proline, BCH altered neither glucose formation nor glutamine and glutamate synthesis, while an accelerated glycerol utilization was accompanied by a small increase in lactate production. In view of the BCH-induced changes in intracellular metabolite levels the acceleration of gluconeogenesis by BCH in the presence of alanine+glycerol+octanoate is probably due to (i) increased uptake of alanine via alanine aminotransferase, (ii) stimulation of phosphoenolpyruvate carboxykinase, a key-enzyme of gluconeogenesis, (iii) rise of glucose-6-phosphatase activity, as well as (iv)activation of the malate-aspartate shuttle resulting in an augmented glycerol utilization for lactate and glucose synthesis.
In isolated rabbit renal kidney-cortex tubules 2 mM glycerol, which is a poor gluconeogenic substrate, does not induce glucose formation in the presence of alanine, while it activates gluconeogenesis on substitution of alanine by aspartate, glutamate or proline. The addition of either 5 mM 3-hydroxybutyrate or 5 mM acetoacetate to renal tubules incubated with alanine + glycerol causes a marked induction of glucose production associated with inhibition of glutamine synthesis. In contrast, the rate of the latter process is not altered by ketones in the presence of glycerol and either aspartate, glutamine or proline despite the stimulation of glucose formation. Acceleration of gluconeogenesis by ketone bodies in the presence of amino acids and glycerol is probably due to (i) stimulation of pyruvate carboxylase activity, (ii) activation of malate-aspartate shuttle as concluded from elevated intracellular levels of malate, aspartate and glutamate, as well as (iii) diminished supply of ammonium for glutamine synthesis from alanine resulting from a decrease in glutamate dehydrogenase activity.
Neural stem cells (NSC) are potentially a promising treatment strategy for neurological diseases. There is a proof that intracerebral grafting of NSC can have restorative effects. Nevertheless, a major obstacle is the induction of a host-derived immune response followed by graft injection. The goal of the study was to analyze the fate of HUCB-NSC transplanted in injured rat brain. Methods: 2×104 HUCB-NSC were tx into corpus callosum of a focal brain injury induced by OUA injection (1 μl/50 nmol) into striatum of adult Wistar rats. After 1, 3, 7 and 14 days brains were removed and analyzed immunocytochemically. Results. One day after HUCB tx, most cells remained in the injection site with few cells migrated to the lesion area. Concomitantly, infi ltration of ED1+ and CD15+ cells with occasional appearance of CD5+ and CD45+ cells was seen. At 3rd day some HUCB-NSC in graft core expressed neuronal (NF-200) or astrocytic (GFAP) markers. Migration of tx cells into the lesion was observed. At 7th day HUCB-NSC were found close to injured area. By 14 days, no viable HUCB-NSC have been noticed with cellular debris in graft core. Limited number of ED1+, CD15+, CD5+ and CD45+ cells were found most likely due to HUCB-NSC rejection. Conclusions. Massive loss of transplanted HUCB-NSC was probably due to post traumatic infl ammation and acute immune reaction of the host in term of cross-species grafting paradigm. Supported by MSHE grant N401 014235 and Fondation Jerome Lejeune scientifi c grant.
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