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The deubiquitinating enzyme UBPy/USP8 interacts with TrkA and inhibits neuronal differentiation in PC12 cells.
In this study we demonstrated that in PC12 cells the TrkA receptor interacts with the deubiquitinating enzyme USP8/UBPy in a NGF-dependent manner, and that it is deubiquitinated in vivo and in vitro by USP8. USP8 overexpression blocked NGF-induced neurites outgrowth while the overexpression of the catalytically inactive mutant USP8/UBPy(C748A) caused a marked increase of cell differentiation. Localization and biochemical experiments have point out that USP8 and TrkA partially co-localize in endosomes after NGF stimulation. Finally we have studied the role played by USP8 on TrkA turnover; using specific siRNA for USP8 we fo...
Source: Experimental Cell Research - February 4, 2015 Category: Cytology Authors: Ceriani M, Amigoni L, D'Aloia A, Berruti G, Martegani E Tags: Exp Cell Res Source Type: research

Molecular mechanism of 2-APB-induced Ca(2+) influx in external acidification in PC12.
In this study, to identify the possible target for the action of 2-APB we examined the pharmacological and molecular properties of [Ca(2+)]i and secretory responses to 2-APB under extracellular low pH conditions. 2-APB dose-dependently induced a [Ca(2+)]i increase and dopamine release, which were greatly enhanced by the external acidification (pH 6.5). [Ca(2+)]i and secretory responses to 2-APB at pH 6.5 were inhibited by the removal of extracellular Ca(2+) and SOC channel blockers such as SK&F96365, La(3+) and Gd(3+). PC12 expressed all SOC channel molecules, Orai 1, Orai 2 and Orai 3. When we used an siRNA system, do...
Source: Experimental Cell Research - March 11, 2014 Category: Cytology Authors: Takahashi K, Yokota M, Ohta T Tags: Exp Cell Res Source Type: research

Ephedrine induced Thioredoxin-1 expression through β-adrenergic receptors/Cyclic AMP/protein kinase A/dopamine- and Cyclic AMP-regulated phosphoprotein signaling pathway.
In this study, we found that Eph induced Trx-1 expression, which was inhibited by propranolol (β-adrenergic receptors inhibitor), but not by phenoxybenzamine (α-adrenergic receptors inhibitor) in rat pheochromocytoma PC12 cells. Moreover, the increase of Trx-1 expression was inhibited by SQ22536 (adenylyl cyclase inhibitor) and H-89 (protein kinase A inhibitor). Interestingly, the effect of Eph on dopamine- and Cyclic AMP-regulated phosphoprotein (DARPP-32) was similar to Trx-1. Thus, the relationship between Trx-1 and DARPP-32 was further studied. The DARPP-32 siRNA significantly reduced Trx-1 expression, but Trx-1 siRN...
Source: Cellular Signalling - February 14, 2013 Category: Cytology Authors: Jia JJ, Zeng XS, Li Y, Ma S, Bai J Tags: Cell Signal Source Type: research

Thioredoxin-1 was Required for CREB Activity by Methamphetamine in Rat Pheochromocytoma Cells.
In this study, we found that Trx-1 was induced by METH in rat pheochromocytoma PC12 cells. Furthermore, PI3K/Akt pathway was involved in METH-induced increase of Trx-1 expression. An increase in phosphorylated cAMP response element-binding protein (CREB) was also observed after exposure of PC12 cells to METH, which was inhibited by a PI3K inhibitor, LY294002. In addition, the siRNA targeted toTrx-1 reduced the level of phosphorylated CREB by METH, suggesting Trx-1 is necessary for increased activity of CREB by METH. The results obtained in this study showed that Trx-1 might play a role in the actions of METH. PMID: 23...
Source: Cellular and Molecular Neurobiology - December 13, 2012 Category: Cytology Authors: Lv T, Wang SD, Bai J Tags: Cell Mol Neurobiol Source Type: research