Saluja A K, Dawra R K, Lerch M M, Steer M L
Harvard Medical School, Harvard Digestive Diseases Center, Boston, Massachusetts.
J Biol Chem. 1992 Jun 5;267(16):11202-7.
In pancreatic acinar cells cholecystokinin and its analogs, caerulein and CCK-JMV-180, stimulate an increase in intracellular free [Ca2+] by releasing Ca2+ from non-mitochondrial intracellular pools. It is generally believed that the caerulein-induced release of Ca2+ is mediated by phospholipase C-catalyzed production of 1,4,5-inositol triphosphate (IP3). In this study we have investigated the source and mechanism of Ca2+ release induced by CCK-JMV-180 using streptolysin O-permeabilized pancreatic acinar cells. Caerulein-stimulated release of Ca2+ was completely blocked by either neomycin, an inhibitor of phospholipase C, or by heparin, an IP3 receptor antagonist. These observations are compatible with the conclusion that caerulein releases Ca2+ from an IP3-sensitive pool. In contrast to caerulein, however, CCK-JMV-180-stimulated release of Ca2+ was not blocked by either neomycin or by heparin, indicating that CCK-JMV-180 releases Ca2+ by mechanisms which do not involve the generation or action of IP3. CCK-JMV-180 stimulated the release of Ca2+ even after the IP3-sensitive pool had been completely emptied by prior exposure to a supramaximally stimulating concentration of IP3 (40 microM). Prestimulation of permeabilized acini with 20 mM caffeine did not abolish the CCK-JMV-180-induced Ca2+ release. These results indicate that CCK-JMV-180 stimulates release of Ca2+ from a hitherto uncharacterized intracellular storage pool which is insensitive to either IP3 or caffeine.
在胰腺腺泡细胞中,胆囊收缩素及其类似物蛙皮素和CCK-JMV-180,通过从非线粒体细胞内钙库释放Ca2+,刺激细胞内游离[Ca2+]增加。一般认为,蛙皮素诱导的Ca2+释放是由磷脂酶C催化生成1,4,5-肌醇三磷酸(IP3)介导的。在本研究中,我们使用链球菌溶血素O通透的胰腺腺泡细胞,研究了CCK-JMV-180诱导的Ca2+释放的来源和机制。磷脂酶C抑制剂新霉素或IP3受体拮抗剂肝素,均可完全阻断蛙皮素刺激的Ca2+释放。这些观察结果与蛙皮素从IP3敏感钙库释放Ca2+的结论相符。然而,与蛙皮素不同,新霉素或肝素均不能阻断CCK-JMV-180刺激的Ca2+释放,这表明CCK-JMV-180通过不涉及IP3生成或作用的机制释放Ca2+。即使在IP3敏感钙库因预先暴露于超最大刺激浓度的IP3(40 microM)而完全排空后,CCK-JMV-180仍能刺激Ca2+释放。用20 mM咖啡因对通透腺泡进行预刺激,并未消除CCK-JMV-180诱导的Ca2+释放。这些结果表明,CCK-JMV-180刺激从一个迄今未被描述的细胞内储存钙库释放Ca2+,该钙库对IP3或咖啡因均不敏感。