Recording of voltage and Ca(2+)-dependent currents in Xenopus oocytes using an intracellular perfusion method.
We describe a method for internal perfusion of Xenopus laevis oocytes that allows control of the composition of intracellular and extracellular solutions, including the possibility of sequential introduction of different substances inside and outside the cell. Using this method, it was possible to record Ca2+ dependent Cl- current and to inhibit it by intracellular perfusion of EGTA-containing solution. With a high BA2+ solution at the external surface of the perfused oocyte, Ba2+ currents through voltage-dependent Ca2+ channels were observed in native and in cardiac RNA-injected oocytes. Finally, a delayed rectifier K+ current was recorded and blocked by internally perfused Cs+ in oocytes injected with mRNA of a cloned (MBK1) K+ channel. The method is expected to be useful for the study of function and modulation of ion channels and transporters in the oocyte, which is an important and widely used model system.
Dascal
N
N
Department of Physiology and Pharmacology, Sackler School of Medicine, Tel Aviv University, Israel.
Chilcott
G
G
Lester
H A
HA
D000818:Animals; D001464:Barium / Q000378:metabolism; D001933:Brain Stem / Q000201:enzymology; D002121:Calcium Channel Blockers / Q000494:pharmacology; D015220:Calcium Channels / Q000378:metabolism*; D003593:Cytoplasm / Q000201:enzymology; D004594:Electrophysiology; D005260:Female; D007124:Immunoenzyme Techniques; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D009206:Myocardium / Q000378:metabolism; D009865:Oocytes / Q000378:metabolism*; D012313:RNA / Q000378:metabolism; D014982:Xenopus laevis
D002121:Calcium Channel Blockers; D015220:Calcium Channels; D001464:Barium; D012313:RNA