Amperometric analysis of exocytosis at chromaffin cells from genetically distinct mice
Amperometric analysis of exocytosis at chromaffin cells from genetically distinct mice
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DOI：
10.1016/s0165-0270(00)00359-9
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发表时间：
2001-01-30
影响因子：
3
通讯作者：
Ewing, AG
中科院分区：
文献类型：
--
作者：
Colliver, TL;Hess, EJ;Ewing, AG
安培法是一种非常强大的技术，可以在单细胞水平上研究(S)特定蛋白质在胞吐中所起的作用。在这项研究中，我们使用安培法研究了从缺陷和摇摇欲坠的突变小鼠中分离的嗜铬细胞胞吐功能的可能变化。假瘤小鼠具有包含突触前蛋白SNAP-25基因的缺失突变，摇摇欲坠的小鼠携带α(1A)亚单位基因突变，该亚单位基因编码P/Q型钙通道的成孔区。虽然从摇摇欲坠的细胞和缺陷性细胞测量的安培数据与野生型对照细胞测量的数据没有显著差异，但在室温和37℃下对野生型嗜铬细胞组进行分析时，发现了显著的差异。由于安培数据固有的巨大变异性，可能无法检测到某些遗传差异导致的释放变化。为了充分利用使用小鼠嗜铬细胞的技术优势，描述了实验指南，这些指南应该最大限度地改变由于遗传差异而导致的释放，并增加检测到安培数据变化的可能性。(C)2001 Elsevier Science B.V.保留所有权利。
Amperometry is a very powerful technique for investigating the role(s) specific proteins play in exocytosis at the single-cell level. In this study, amperometry has been used to investigate possible changes in exocytosis at chromaffin cells isolated from coloboma and tottering mutant mice. Coloboma mice possess a deletion mutation that encompasses the gene for the presynaptic protein SNAP-25 and tottering mice carry a mutation of the alpha (1A) subunit gene, which encodes the pore-forming region of P/Q-type calcium channels. Although amperometric data measured from tottering and coloboma cells are not significantly different from that measured at wild-type control cells, significant differences are found when groups of wild-type chromaffin cells an analyzed at room temperature and at 37 degreesC. Due to the large variability inherent to amperometric data, it is possible that changes in release resulting from some genetic differences cannot be detected. To fully exploit the technical advantages of using mouse chromaffin cells, experimental guidelines are described which should maximize changes in release resulting from genetic differences and increase the likelihood of detecting a change in amperometric data. (C) 2001 Elsevier Science B.V. All rights reserved.