Fogarty K E, Kidd J F, Tuft D A, Thorn P
Department of Pharmacology, Tennis Court Road, Cambridge CB2 1QJ, UK.
J Physiol. 2000 Aug 1;526 Pt 3(Pt 3):515-26. doi: 10.1111/j.1469-7793.2000.t01-1-00515.x.
In secretory epithelial cells, complex patterns of Ca2+ signals regulate physiological processes. How these patterns are generated is still not fully understood. In particular, the basis of global Ca2+ waves is not clear. We have studied regional differences in InsP3-evoked Ca2+ release in single mouse pancreatic acinar cells, using high-speed (approximately 90 frames s-1), high-sensitivity Ca2+ imaging combined with rapid (10 ms) spot photolysis (2 micrometer diameter) of caged InsP3. Within a single region we measured Ca2+ response latency and rate of rise to construct an InsP3 dose-response relationship. Spot InsP3 liberation in the secretory pole region consistently elicited a dose-dependent, rapid release of Ca2+. Spot InsP3 liberation in the basal pole region of approximately 50% of cells elicited a similar dose-response relationship but with a lower apparent InsP3 affinity than in the secretory pole. In the other cells, basal pole InsP3 liberation did not elicit active Ca2+ release, even at the highest stimulus intensities we employed, although these same cells did respond when the stimulus spot was moved to different regions. We conclude that in the basal pole active sites of rapid Ca2+ release have a lower functional affinity for InsP3 than those in the secretory pole and are spread out in discrete sites across the basal pole. These properties explain the propagation of Ca2+ waves across the basal pole that are only observed at higher stimulus levels.
在分泌上皮细胞中,复杂的Ca2+信号模式调节着生理过程。这些模式是如何产生的仍未完全清楚。特别是,全局Ca2+波的基础尚不清楚。我们利用高速(约90帧/秒)、高灵敏度的Ca2+成像技术,结合对笼锁肌醇三磷酸(InsP3)进行快速(10毫秒)光斑光解(直径2微米),研究了单个小鼠胰腺腺泡细胞中InsP3诱发的Ca2+释放的区域差异。在单个区域内,我们测量了Ca2+反应潜伏期和上升速率,以构建InsP3剂量反应关系。在分泌极区域进行光斑InsP3释放始终能引发剂量依赖性的Ca2+快速释放。在约50%的细胞的基底极区域进行光斑InsP3释放引发了类似的剂量反应关系,但与分泌极相比,其表观InsP3亲和力较低。在其他细胞中,即使在我们采用的最高刺激强度下,基底极InsP3释放也不会引发Ca2+的主动释放,尽管当刺激光斑移至不同区域时,这些相同的细胞确实会产生反应。我们得出结论,在基底极,快速Ca2+释放的活性位点对InsP3的功能亲和力低于分泌极的位点,并且在基底极以离散位点分布。这些特性解释了仅在较高刺激水平下才观察到的Ca2+波在基底极的传播。