Imaging of chnages in intracellular ion distribution induced by receptor stimulation in parotid aciner cells

1. 毒蕈碱受体刺激诱导Ca^<2+>波的成像。用calbachol （CCh）刺激负载fura-2的大鼠腮腺腺泡细胞。在腺泡细胞的顶极开始增加，然后以Ca^<2+>波向基底外侧区迅速扩散。闪光光解1P3诱导皂苷渗透腮腺腺泡细胞Ca^<2+>波。通透性腺泡细胞被膜结合的Ca^<2+>指示剂钙绿C_<18>标记，并受到笼状IP光解的刺激，细胞内储存的Ca^<2+>从顶端区域释放，向基底外侧区域扩散。通透性腮腺腺泡细胞内Ca^<2+>储存释放的C^<2+>的成像。使用荧光指示剂magu -fura-2和钙绿C_<18>监测细胞内Ca^<2+>库的Ca^<2+>释放。IP_3的应用引起Ca^<2+>的快速释放，但环adp核糖、咖啡因和ryanodine均不刺激Ca^<2+>的释放。异丙肾上腺素对胞浆Ca^<2+>分布的影响。使用能产生最大淀粉酶释放的1 -甲基异丙肾上腺素浓度，没有引起胞浆Ca^<2+>分布的任何变化。CCh对细胞质CI分布的影响。CCh引起胞浆内CF浓度迅速下降。这一变化与[Ca^<2+>]的增加有关。

1. Imaging of Ca^<2+> waves induced by muscarinic receptor stimulation. The fura-2 loaded rat parotid acinar cells were stimulated with calbachol (CCh). The increase in was initiated in the apical pole of the acinar cells and then rapidly spread as a Ca^<2+> wave towards the basolateral region.2. The Ca^<2+> wave in saponin-permeabilized parotid acinar cells evoked by flash photolysis of caged 1P3. The permeabilized acinar cells were labelled with the membrane-bound Ca^<2+> indicator Calcium Green C_<18> and stimulated by the photolysis of caged IP The Ca^<2+> release from intracellular stores started at the apical region and spread towards the basolateral region.3. Imaging of C^<2+> release from intracellular Ca^<2+> stores in permeabilized parotid acinar cells. The Ca^<2+> release from intracellular Ca^<2+> stores was monitored using the fluorescent indicators Mag-fura-2 and Calcium Green C_<18> App1ication of IP_3 caused a rapid release of Ca^<2+>, but none of cyclic ADP-ribose, caffeine or ryanodine stimulated Ca^<2+> release.4. Effect of isoproterenol on cytosolic Ca^<2+> distribution. Application of 1 muMisoproterenol, a concentration producing the maximum amylase release, did not cause any change in cytosolic Ca^<2+> distribution.5. Effect of CCh on cytosolic CI distribution. CCh caused a rapid dcrease in cytosolic CF concentration. This change was corelated with the increase in [Ca^<2+>].