Two distinct mechanisms underlie the stimulation of neurotransmitter release by phorbol esters in clonal rat pheochromocytoma PC12 cells

被引:40
作者
Iwasaki, S
Kataoka, M
Sekiguchi, M
Shimazaki, Y
Sato, K
Takahashi, M [1 ]
机构
[1] Mitsubishi Kasei Inst Life Sci, Tokyo 1948511, Japan
[2] Univ Tokyo, Grad Sch Arts & Sci, Dept Life Sci Biol, Meguro Ku, Tokyo 1538902, Japan
关键词
neurotransmitter release; phorbol ester; phosphorylation; PKC; SNAP-25;
D O I
10.1093/oxfordjournals.jbchem.a022768
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Phorbol ester treatment induces the phosphorylation of SNAP-25 at Ser(187) and the potentiation. of Ca2+-induced dopamine (DA) and acetylcholine (Ach) release from PC12 cells. In order to evaluate the functional consequences of phosphorylation, quantitative analysis was carried out using an. anti-phosphopeptide antibody that specifically recognizes SNAP-25 phosphorylated at Ser(187), DA and ACh release, assayed in low-K+ as well as high-K+ solution, increased by treating the cells with phorbol-12-myristate-13-acetate (PMA); however, the stimulation of high-K+-dependent release occurred at lower concentrations and with shorter exposures to PMA than that of the basal release in low-K+-solution, The PMA-induced phosphorylation of SNAP-25 did not correlate with the potentiation of high-K+-dependent neurotransmitter release. The potentiation of high-K+-dependent DA release by phorbol 12,13-diacetate (PDA), a water soluble phorbol ester, almost completely disappeared within 1 min after washing PDA in the presence of okadaic acid, conditions under which the phosphorylation of SNAP-25 persisted for at least 15 min, PMA-induced phosphorylation of SNAP-25 was inhibited by staurosporine, however, the potentiation of high-K+-dependent PA release was suppressed only partially. These results indicate that protein kinase activation does not account for a large fraction of the phorbol ester-induced potentiation of depolarization-dependent neurotransmitter release from PC12 cells.
引用
收藏
页码:407 / 414
页数:8
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