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MessagePosté le: Mar 30 Sep - 09:05 (2014) Sujet du message: On this way a macroneuron population regulates the number o Répondre en citant

 This was confirmed utilizing supplier INNO-406 MEFs from Evc2 null mice exactly where Evc is present but does not localize to the base of main cilia. Co dependent loca lization of Evc and Evc2 could describe why mutations in either gene consequence in an indistinguishable patient pheno sort. Given that each Evc and Evc2 possess transmem brane domains, observing them about the cilium led us to question whether or not they span the cilia membrane and, if so, their orientation in the membrane. We addressed this by comparison of immunofluorescence on permea bilized and non permeabilized cells with antibodies gen erated against peptides N terminal and C terminal with the transmembrane domain of Evc2. These confirmed that Evc2 spans the cilia membrane and that the N terminal portion is extracellular.

Evc includes a predicted signal anchor sequence supplier Lapatinib and comparison of immuno fluorescence signal on permeabilized and non permeabi lized cells demonstrated that Evc is intracellular. In addition, the regions of Evc and Evc2 that we have demonstrated to interact are each intracellular. A query that arises when a Hh signalling defect outcomes from reduction of the cilia protein is whether this defect is due to a cilia abnormality or as a result of distinct modulation of Hh signal transduction. Ultrastructural analysis of cilia of Evc mutant cells doesn't uncovered any abnormalities suggesting that the mutant phenotype won't end result from defective cilia but rather that Evc is actually a distinct modi fier of Hh signal transduction. Moreover on the cilia localization, we detected Evc2 in MEF nuclear extracts by Western blotting.

We detected Evc in cytoplasmic but not in nuclear extracts suggesting that Evc won't enter the nucleus, though we are unable to exclude the likelihood that Evc is present at undetectable ranges in the Lonafarnib 価格 nucleus. Evc and Evc2 were not observed within the nucleus by immunostaining, but a diffuse nuclear distribution may very well be tough to distin guish from background staining. Also, overexpressed proteins weren't observed within the nucleus when expressed alone or when co expressed main us to conclude that more variables are required to trans port Evc2 into the nucleus, a method that could rely on activation with the Hh pathway. The presence of Evc2 both during the ciliary membrane and in the nucleus is intriguing.

There exists a precedent for proteolytic processing and translocation for the nucleus of your C terminal region of the cilia membrane protein in polycystin one. Following cleavage, the C terminal peptide of PC1 moves to your nucleus in which it associates with Stat6 and p100 to activate gene tran scription. The nuclear localization and also the N terminal extracellular portion of Evc2 may indicate an analogous position for Evc2 from the regulation of Hh target gene tran scription in response to extracellular signals. Among the a lot of latest surprises regarding Hh signal ling was the getting the Gli transcription aspects localize to cilia also as the nucleus. Even more experi ments are necessary to determine no matter whether Ellis van Cre veld proteins move in conjunction with Gli proteins or by way of an independent pathway. Conclusions We now have previously shown that Evc is a good modu lator of Ihh signalling with the growth plate acting at or downstream of Smo.


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MessagePosté le: Mar 30 Sep - 09:05 (2014) Sujet du message: Publicité

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