通过笼锁Ca2+的闪光光解刺激单个大鼠垂体细胞分泌的毫秒级研究。
Millisecond studies of secretion in single rat pituitary cells stimulated by flash photolysis of caged Ca2+.
作者信息
Thomas P, Wong J G, Almers W
机构信息
Department of Physiology and Biophysics, University of Washington, Seattle 98195.
出版信息
EMBO J. 1993 Jan;12(1):303-6. doi: 10.1002/j.1460-2075.1993.tb05657.x.
Abstract
To study the final steps in the secretory pathway of rat pituitary melanotrophs, we have monitored changes in cell surface area due to exocytosis after flash photolysis of caged Ca2+. A step increase in cytosolic [Ca2+] to 45-125 microM triggers three phases of exocytic secretion. A small cohort of a few hundred vesicles is exocytosed in 40 ms in a secretory burst with a peak rate of 17,000 vesicles/s. Next, 2700 more vesicles are released in a slower phase that is complete within 400-1000 ms. Finally, vesicles continue to be released slowly (500 vesicles/s) for > 8s. The approach described provides a way to identify and monitor the final steps in the secretory pathway at millisecond resolution. That a small portion of secretory vesicles can be released much faster than all others suggests that these vesicles are functionally equivalent to those at the presynaptic active zone of a neuron. Their release would be fast enough to be temporally correlated with single action potentials.
摘要
为了研究大鼠垂体黑素细胞分泌途径的最后步骤，我们在笼锁Ca2+的闪光光解后监测了由于胞吐作用导致的细胞表面积变化。胞质[Ca2+]瞬间增加到45 - 125微摩尔引发了三个阶段的胞吐分泌。几百个小囊泡在40毫秒内以17,000个囊泡/秒的峰值速率在分泌爆发中胞吐。接下来，另外2700个囊泡在400 - 1000毫秒内以较慢的阶段释放。最后，囊泡继续缓慢释放（500个囊泡/秒）超过8秒。所描述的方法提供了一种以毫秒分辨率识别和监测分泌途径最后步骤的方法。一小部分分泌囊泡能够比其他所有囊泡释放得快得多，这表明这些囊泡在功能上等同于神经元突触前活动区的囊泡。它们的释放速度足够快，能够在时间上与单个动作电位相关联。
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