Amperometric analysis of exocytosis at chromaffin cells from genetically distinct mice

被引:29
作者
Colliver, TL
Hess, EJ
Ewing, AG [1 ]
机构
[1] Penn State Univ, Dept Chem, Davey Lab 152, University Pk, PA 16802 USA
[2] Penn State Univ, Coll Med, Dept Anat & Neurosci, Hershey, PA 17033 USA
基金
美国国家卫生研究院;
关键词
amperometry; single cell; exocytosis; mouse chromaffin cells; tottering; coloboma;
D O I
10.1016/S0165-0270(00)00359-9
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:95 / 103
页数:9
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