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Functional inhibition of native volume‐sensitive outwardly rectifying anion channels in muscle cells and Xenopus oocytes by anti‐ClC‐3 antibody
Author(s) -
Duan Dayue,
Zhong Juming,
Hermoso Marcela,
Satterwhite Christina M.,
Rossow Charles F.,
Hatton William J.,
Horowitz Burton,
Hume Joseph R.
Publication year - 2001
Publication title -
the journal of physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.802
H-Index - 240
eISSN - 1469-7793
pISSN - 0022-3751
DOI - 10.1111/j.1469-7793.2001.0437i.x
Subject(s) - xenopus , intracellular , polyclonal antibodies , epitope , endogeny , chemistry , microbiology and biotechnology , cardiac muscle , antibody , biology , biochemistry , anatomy , immunology , gene
1 Intracellular dialysis of NIH/3T3 cells with a commercially available anti‐ClC‐3 polyclonal antibody (Ab) for ≈30 min completely inhibited expressed guinea‐pig ClC‐3 currents ( I gpClC‐3 ), while intracellular dialysis with antigen‐preabsorbed anti‐ClC‐3 Ab failed to affect I gpClC‐3 . 2 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. 3 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.