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J Neurosci Methods. 2010 May 15;188(2):280-6. doi: 10.1016/j.jneumeth.2010.03.006. Epub 2010 Mar 15.

Oocyte triplet pairing for electrophysiological investigation of gap junctional coupling.

Author information

1
Dept of Neurobiology and Developmental Sciences, University of Arkansas for Medical Sciences, Center for Translational Neuroscience, Little Rock, AR 72205, United States. abdallah@hayar.net

Abstract

Gap junctions formed by expressing connexin subunits in Xenopus oocytes provide a valuable tool for revealing the gating properties of intercellular gap junctions in electrically coupled cells. We describe a new method that consists of simultaneous triple recordings from 3 apposed oocytes expressing exogenous connexins. The advantages of this method are that in one single experiment, 1 oocyte serves as control while a pair of oocytes, which have been manipulated differently, may be tested for different gap junctional properties. Moreover, we can study simultaneously the gap junctional coupling of 3 different pairs of oocytes in the same preparation. If the experiment consists of testing the effect of a single drug, this approach will reduce the time required, as background coupling in control pairs of oocytes does not need to be measured separately as with the conventional 2 oocyte pairing. The triplet approach also increases confidence that any changes seen in junctional communication are due to the experimental treatment and not variation in the preparation of oocytes or execution of the experiment. In this study, we show the example of testing the gap junctional properties among 3 oocytes, 2 of which are expressing rat connexin36.

PMID:
20230857
PMCID:
PMC2859429
DOI:
10.1016/j.jneumeth.2010.03.006
[Indexed for MEDLINE]
Free PMC Article

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