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

1.电刺激M受体诱发的Ca~(2+)&Gt~(2+)波的成像用钙化胆碱(CCH)刺激Fura-2负载的大鼠腮腺腺泡细胞。在腺泡细胞的顶端开始增加，然后以钙波的形式迅速向基外区扩散。笼子1P3闪光光解引起的皂苷通透性腮腺腺泡细胞中的钙波。用膜结合的钙指示剂钙绿C 18&gt；标记通透性的腺泡细胞，在笼状IP光解的刺激下，细胞内的钙离子从细胞顶端开始释放，并向基底区扩散。C^&lt；2+&gt；从细胞内释放C^&lt；2+&gt；储存在通透性腮腺腺泡细胞中的成像。用荧光指示剂Mag-Fura-2和钙绿C_(18&gt；)监测细胞内Ca~(2+)的释放。IP_3的应用引起Ca~(2+)的快速释放，但环状ADP-核糖、咖啡因和兰诺定不能刺激Ca~(2+)的释放。异丙肾上腺素对细胞内钙离子分布的影响施加产生最大淀粉酶释放浓度的1-米索丙肾上腺素，没有引起胞内Ca~(2+)&gt；分布的任何变化。CCH对胞浆CI分布的影响CCH引起细胞内CF浓度迅速下降。这一变化与[Ca^&lt；2+&gt；]的升高有关。

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+>].