dbcAMP
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Cellular differentiation is the process, by which a cell changes from one cell type to another, preferentially to the more specialized one. Calcium fluxes play an important role in this action. Differentiated NG108-15 or PC12 cells serve as models for studying neuronal pathways. NG108-15 cell line is a reliable model of cholinergic neuronal cells. These cells differentiate to a neuronal phenotype due to the dibutyryl cAMP (dbcAMP) treatment. We have shown that a slow sulfide donor - GYY4137 - can also act as a differentiating factor in NG108-15 cell line. Calcium is an unavoidable ion required in NG108-15 cell differentiation by both, dbcAMP and GYY4137, since cultivation in EGTA completely prevented differentiation of these cells. In this work we focused primarily on the role of reticular calcium in the process of NG108-15 cell differentiation. We have found that dbcAMP and also GYY4137 decreased reticular calcium concentration by different mechanisms. GYY4137 caused a rapid decrease in type 2 sarco/endoplasmic calcium ATPase (SERCA2) mRNA and protein, which results in lower calcium levels in the endoplasmic reticulum compared to the control, untreated group. The dbcAMP revealed rapid increase in expression of the type 3 IP3 receptor, which participates in a calcium clearance from the endoplasmic reticulum. These results point to the important role of reticular calcium in a NG108-15 cell differentiation.
- MeSH
- buněčná diferenciace účinky léků MeSH
- dibutyryl cyklický AMP aplikace a dávkování MeSH
- inositol-1,4,5-trisfosfát - receptory metabolismus MeSH
- messenger RNA metabolismus MeSH
- morfoliny aplikace a dávkování MeSH
- myši inbrední BALB C MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- neurony účinky léků fyziologie MeSH
- organothiofosforové sloučeniny aplikace a dávkování MeSH
- sarkoplazmatická Ca2+-ATPáza metabolismus MeSH
- sulfan aplikace a dávkování MeSH
- vápník metabolismus MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
The aim of this study was to investigate the involvement of the serine/threonine protein kinase AKT (also called protein kinase B) in the control of meiosis of porcine denuded oocytes (DOs) matured in vitro. Western blot analysis revealed that the two principal AKT phosphorylation sites, Ser473 and Thr308, are phosphorylated at different stages of meiosis. In freshly isolated germinal vesicle (GV)-stage DOs, Ser473 was already phosphorylated. After the onset of oocyte maturation, the intensity of the Ser473 phosphorylation increased, however, which declined sharply when DOs underwent GV breakdown (GVBD) and remained at low levels in metaphase I- and II-stage (MI- and MII-stage). In contrast, phosphorylation of Thr308 was increased by the time of GVBD and reached maximum at MI-stage. A peak of AKT activity was noticed around GVBD and activity of AKT declined at MI-stage. To assess the role of AKT during meiosis, porcine DOs were cultured in 50 microM SH-6, a specific inhibitor of AKT. In SH-6-treated DOs, GVBD was not inhibited; on the contrary, a significant acceleration of meiosis resumption was observed. The dynamics of the Ser473 phosphorylation was not affected; however, phosphorylation of Thr308 was reduced, AKT activity was diminished at the time of GVBD, and meiotic progression was arrested in early MI-stage. Moreover, the activity of the cyclin-dependent kinase 1 (CDK1) and MAP kinase declined when SH-6-treated DOs underwent GVBD, indicating that AKT activity is involved in the regulation of CDK1 and MAP kinase. These results suggest that activity of AKT is not essential for induction of GVBD in porcine oocytes but plays a substantial role during progression of meiosis to MI/MII-stage.
- MeSH
- aktivace enzymů účinky léků fyziologie MeSH
- buněčné kultury MeSH
- dibutyryl cyklický AMP farmakologie MeSH
- fosfatidylinositoly farmakologie MeSH
- fosforylace účinky léků MeSH
- inhibitory enzymů farmakologie MeSH
- kultivované buňky MeSH
- meióza fyziologie MeSH
- onkogenní protein v-akt antagonisté a inhibitory metabolismus fyziologie MeSH
- oocyty účinky léků metabolismus fyziologie MeSH
- oogeneze účinky léků fyziologie MeSH
- prasata metabolismus MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- chemická stimulace MeSH
- dibutyryl cyklický AMP farmakologie MeSH
- dijodtyrosin metabolismus MeSH
- glukosafosfáty farmakologie MeSH
- glutathion farmakologie MeSH
- izotopy jodu MeSH
- játra metabolismus MeSH
- jod metabolismus MeSH
- krysa rodu rattus MeSH
- kultivační techniky MeSH
- NADP farmakologie MeSH
- prasata MeSH
- radioizotopy jodu MeSH
- štítná žláza metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
