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.
Hayar
Abdallah
A
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
Charlesworth
Amanda
A
Garcia-Rill
Edgar
E
D000818:Animals; D002462:Cell Membrane / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D017630:Connexins / Q000235:genetics / Q000378:metabolism; D004558:Electric Stimulation; D004594:Electrophysiology / Q000295:instrumentation / Q000379:methods*; D005260:Female; D017629:Gap Junctions / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D005822:Genetic Vectors; D008564:Membrane Potentials / Q000235:genetics; D009446:Neurobiology / Q000295:instrumentation / Q000379:methods*; D009865:Oocytes / Q000378:metabolism* / Q000648:ultrastructure; D018408:Patch-Clamp Techniques / Q000295:instrumentation / Q000379:methods; D051381:Rats; D017207:Rats, Sprague-Dawley; D014162:Transfection; D014982:Xenopus laevis; D000097004:Gap Junction delta-2 Protein
D017630:Connexins; D000097004:Gap Junction delta-2 Protein