Functional inhibition of native volume-sensitive outwardly rectifying anion channels in muscle cells and Xenopus oocytes by anti-ClC-3 antibody.
Intracellular dialysis of NIH/3T3 cells with a commercially available anti-ClC-3 polyclonal antibody (Ab) for approximately 30 min completely inhibited expressed guinea-pig ClC-3 currents (IgpClC-3), while intracellular dialysis with antigen-preabsorbed anti-ClC-3 Ab failed to affect IgpClC-3. Anti-ClC-3 Ab was used as a selective probe to examine the relationship between endogenous ClC-3 expression and native volume-sensitive outwardly rectifying anion channels (VSOACs) in guinea-pig cardiac cells, canine pulmonary arterial smooth muscle cells (PASMCs) and Xenopus laevis oocytes. Intracellular dialysis or injection of anti-ClC-3 Ab abolished native VSOAC function in cardiac cells and PASMCs and significantly reduced VSOACs in oocytes. In contrast, native VSOAC function was unaltered by antigen-preabsorbed anti-ClC-3 Ab. It is suggested that endogenous ClC-3 represents a major molecular entity responsible for native VSOACs in cardiac and smooth muscle cells and Xenopus oocytes. Anti-ClC-3 Ab should be a useful experimental tool to directly test the relationship between endogenous ClC-3 expression and native VSOAC function, and help resolve existing controversies related to the regulation and physiological role of native VSOACs in a wide variety of different cells.
Duan
D
D
Centre of Biomedical Research Excellence, Department of Physiology and Cell Biology, University of Nevada School of Medicine, Reno, NV 89557-0046, USA.
Zhong
J
J
Hermoso
M
M
Satterwhite
C M
CM
Rossow
C F
CF
Hatton
W J
WJ
Yamboliev
I
I
Horowitz
B
B
Hume
J R
JR
D016475:3T3 Cells; D000818:Animals; D000838:Anions / Q000378:metabolism*; D000906:Antibodies / Q000494:pharmacology*; D018118:Chloride Channels / Q000276:immunology*; D005260:Female; D007267:Injections; D007473:Ion Channels / Q000037:antagonists & inhibitors*; D051379:Mice; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism*; D009206:Myocardium / Q000166:cytology / Q000378:metabolism; D009865:Oocytes / Q000378:metabolism*; D014982:Xenopus laevis
D000838:Anions; D000906:Antibodies; D018118:Chloride Channels; C115376:ClC-3 channel; D007473:Ion Channels