Tinel H, Cancela J M, Mogami H, Gerasimenko J V, Gerasimenko O V, Tepikin A V, Petersen O H
Medical Research Council Secretory Control Research Group, Physiological Laboratory, University of Liverpool, Liverpool L69 3BX, UK.
EMBO J. 1999 Sep 15;18(18):4999-5008. doi: 10.1093/emboj/18.18.4999.
Agonist-evoked cytosolic Ca(2+) spikes in mouse pancreatic acinar cells are specifically initiated in the apical secretory pole and are mostly confined to this region. The role played by mitochondria in this process has been investigated. Using the mitochondria-specific fluorescent dyes MitoTracker Green and Rhodamine 123, these organelles appeared as a bright belt concentrated mainly around the secretory granule area. We tested the effects of two different types of mitochondrial inhibitor on the cytosolic Ca(2+) concentration using simultaneous imaging of Ca(2+)-sensitive fluorescence (Fura 2) and electrophysiology. When carbonyl cyanide m-chlorophenylhydrazone (CCCP) was applied in the presence of the Ca(2+)-releasing messenger inositol 1,4, 5-trisphosphate (IP(3)), the local repetitive Ca(2+) responses in the granule area were transformed into a global rise in the cellular Ca(2+) concentration. In the absence of IP(3), CCCP had no effect on the cytosolic Ca(2+) levels. Antimycin and antimycin + oligomycin had the same effect as CCCP. Active mitochondria, strategically placed around the secretory pole, block Ca(2+) diffusion from the primary Ca(2+) release sites in the granule-rich area in the apical pole to the basal part of the cell containing the nucleus. When mitochondrial function is inhibited, this barrier disappears and the Ca(2+) signals spread all over the cytosol.
激动剂诱发的小鼠胰腺腺泡细胞胞质钙离子峰在顶端分泌极特异性起始,并大多局限于该区域。研究了线粒体在此过程中所起的作用。使用线粒体特异性荧光染料MitoTracker Green和罗丹明123,这些细胞器呈现为一条明亮的带,主要集中在分泌颗粒区域周围。我们通过同时对钙离子敏感荧光(Fura 2)进行成像和电生理学方法,测试了两种不同类型的线粒体抑制剂对胞质钙离子浓度的影响。当在钙离子释放信使肌醇1,4,5-三磷酸(IP(3))存在的情况下应用羰基氰化物间氯苯腙(CCCP)时,颗粒区域的局部重复性钙离子反应转变为细胞内钙离子浓度的整体升高。在没有IP(3)的情况下,CCCP对胞质钙离子水平没有影响。抗霉素以及抗霉素+寡霉素与CCCP具有相同的作用。活跃的线粒体策略性地分布在分泌极周围,阻止钙离子从顶端富含颗粒区域的主要钙离子释放位点扩散到含有细胞核的细胞基部。当线粒体功能受到抑制时,这种屏障消失,钙离子信号扩散到整个细胞质中。