XB-ART-41298J Neurosci Methods May 15, 2010; 188 (2): 280-6.
Oocyte triplet pairing for electrophysiological investigation of gap junctional coupling.
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.
PubMed ID: 20230857
PMC ID: PMC2859429
Article link: J Neurosci Methods
Grant support: ,RR020146 NCRR NIH HHS , DC07123 NIDCD NIH HHS , NS020246 NINDS NIH HHS , P20 RR020146-06 NCRR NIH HHS , R01 DC007123-06 NIDCD NIH HHS , R01 NS020246-23 NINDS NIH HHS , R01 NS020246-25 NINDS NIH HHS , P20 RR020146 NCRR NIH HHS , R01 DC007123 NIDCD NIH HHS , R01 NS020246 NINDS NIH HHS
Genes referenced: gjd2