Modulation of histamine-induced Ca2+ release by protein kinase C -: Effects on cytosolic and mitochondrial [Ca2+] peaks
Modulation of histamine-induced Ca2+ release by protein kinase C -: Effects on cytosolic and mitochondrial [Ca2+] peaks
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DOI：
10.1074/jbc.m308378200
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发表时间：
2003-12-12
影响因子：
4.8
通讯作者：
Alvarez, J
中科院分区：
文献类型：
--
作者：
Montero, M;Lobatón, CD;Alvarez, J
在HeLa细胞中，组胺诱导肌醇1，4，5-三磷酸（InsP（3））的产生和内质网（ER）中Ca 2+的释放。Ca 2+释放通常是双相的，具有快速和短暂的初始阶段，随后是慢得多且延长的阶段。在蛋白激酶C（PKC）抑制剂（包括星形孢菌素和特异性抑制剂GF 109203 X和Ro-31-8220）的存在下，快速期持续直到ER完全排空。相反，佛波醇12，13-二丁酸酯处理抑制Ca 2+释放。Staurosporine对InsP（3）诱导的透化细胞中Ca 2+释放没有影响，也没有改变组胺诱导的InsP（3）产生。这些数据表明，组胺诱导Ca ~（2+）释放，并在短的滞后激活PKC下调它。同样，组胺诱导的Ca ~（2+）振荡的幅度增加，频率降低，在存在PKC抑制剂。我们还表明，线粒体[Ca 2 +]比胞浆[Ca 2 +]对PKC调节诱导的ER-Ca 2+释放的变化更敏感。PKC抑制剂使组胺诱导的线粒体[Ca 2 +]峰增加4倍，但仅使胞质[Ca 2 +]峰增加20%。相反，PKC激活抑制线粒体[Ca ~（2+）]峰的90%和胞浆的一个只有50%。类似地，PKC抑制剂与线粒体Ca 2+单向转运体激活剂SB 202190的组合导致线粒体[Ca 2 +]峰的急剧增加，对胞质的影响很小。这表明PKC抑制剂激活ER-Ca 2+释放可能参与星形孢菌素诱导的细胞凋亡。此外，这些机制允许在细胞刺激期间灵活和独立地调节胞质和线粒体[Ca 2 +]。
In HeLa cells, histamine induces production of inositol 1,4,5-trisphosphate (InsP(3)) and release of Ca2+ from the endoplasmic reticulum (ER). Ca2+ release is typically biphasic, with a fast and brief initial phase, followed by a much slower and prolonged one. In the presence of inhibitors of protein kinase C (PKC), including staurosporine and the specific inhibitors GF109203X and Ro-31-8220, the fast phase continued until the ER became fully empty. On the contrary, treatment with phorbol 12,13-dibutyrate inhibited Ca2+ release. Staurosporine had no effect on InsP(3)-induced Ca2+ release in permeabilized cells and did not modify either histamine-induced InsP(3) production. These data suggest that histamine induces Ca2+ release and with a short lag activates PKC to down-regulate it. Consistently, Ca2+ oscillations induced by histamine were increased in amplitude and decreased in frequency in the presence of PKC inhibitors. We show also that mitochondrial [Ca2+] was much more sensitive to changes in ER-Ca2+ release induced by PKC modulation than cytosolic [Ca2+]. PKC inhibitors increased the histamine-induced mitochondrial [Ca2+] peak by 4-fold but increased the cytosolic [Ca2+] peak only by 20%. On the contrary, PKC activation inhibited the mitochondrial [Ca2+] peak by 90% and the cytosolic one by only 50%. Similarly, the combination of PKC inhibitors with the mitochondrial Ca2+ uniporter activator SB202190 led to dramatic increases in mitochondrial [Ca2+] peaks, with little effect on cytosolic ones. This suggests that activation of ER-Ca2+ release by PKC inhibitors could be involved in apoptosis induced by staurosporine. In addition, these mechanisms allow flexible and independent regulation of cytosolic and mitochondrial [Ca2+] during cell stimulation.