Amperometric analysis of exocytosis at chromaffin cells from genetically distinct mice

Thomas L. Collivera, Ellen J. Hess, Andrew G. Ewing

Research output: Contribution to journalArticlepeer-review

Abstract

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 α1A 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 are analyzed at room temperature and at 37°C. 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.

Original languageEnglish (US)
Pages (from-to)95-103
Number of pages9
JournalJournal of Neuroscience Methods
Volume105
Issue number1
DOIs
StatePublished - Jan 30 2001

All Science Journal Classification (ASJC) codes

  • Neuroscience(all)

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