Measurement of [Ca2+]i in smooth muscle strips using front-surface fluorimetry.
Measurement of [Ca2+]i in smooth muscle strips using front-surface fluorimetry.
复制标题
使用前表面荧光测定法测量平滑肌条中的[Ca2]i。
DOI：
10.1007/978-1-62703-086-1_12
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发表时间：
1999
影响因子：
--
通讯作者：
H. Kanaide
中科院分区：
文献类型：
--
作者：
H. Kanaide
胞质Ca2+浓度（[Ca2+]i）的变化在平滑肌细胞收缩的调节中起主要作用。然而，在收缩或松弛的时间过程中，[Ca2+]陆地张力之间的关系表现出时间变化。在给定的[Ca2+]i变化的张力发展的程度也取决于收缩和松弛的类型。因此，为了确定平滑肌收缩和舒张调节的分子和细胞机制，同时测量[Ca2+]陆地张力是必要的。本章提供了前表面荧光法技术的基本原理，以及使用fura-2或Fura-PE3作为荧光Ca2+指示剂同时测量平滑肌组织中[Ca2+]离子张力的方案和提示。在平滑肌中加载足量的Ca2+指示剂对于以最小的光学伪影成功测量[Ca2+]至关重要。该方案给出了我们在各种平滑肌组织中Ca2+指标加载的实践。
Changes in the cytosolic Ca2+concentrations ([Ca2+]i) play a primary role in the regulation of the contraction of smooth muscle cells. However, the relationship between [Ca2+]iand tension exhibits a temporal change during the time course of contraction or relaxation. The extent of the tension development for a given change in [Ca2+]ialso varies depending on the type of contraction and relaxation. Therefore, it is essential to measure [Ca2+]iand tension simultaneously in order to determine the molecular and cellular mechanisms in both the regulation of contraction and relaxation of smooth muscle. This chapter provides the basic principles of the technique of front-surface fluorimetry as well as the protocols and tips for the simultaneous measurement of [Ca2+]iand tension in the smooth muscle tissues with use of fura-2 or Fura-PE3 as fluorescent Ca2+indicators. The loading of sufficient amount of the Ca2+indicators in smooth muscles is essential for the successful measurement of [Ca2+]iwith minimum optical artifacts. The protocol gives our practice for the loading of the Ca2+indicators in various smooth muscle tissues.