Open AccessCloning and Characterization of a Calcium-Activated Chloride Channel in Rat Uterus1
Author(s) -
Junfang Song,
Xiaojuan Zhang,
Zhi Qi,
Gang Sun,
Shaopeng Chi,
Ziyan Zhu,
Jiangong Ren,
Zhaojun Qiu,
Liu Ka,
Leslie Myatt,
Z. Runlin
Publication year - 2009
Publication title -
biology of reproduction
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.366
H-Index - 180
eISSN - 1529-7268
pISSN - 0006-3363
DOI - 10.1095/biolreprod.108.071258
Subject(s) - biology , niflumic acid , microbiology and biotechnology , chloride channel , ionomycin , uterus , chinese hamster ovary cell , endocrinology , transfection , complementary dna , medicine , biochemistry , receptor , gene , intracellular
In a search for genes involved in regulation of uterine contractility, we cloned a novel calcium-activated chloride channel gene, named rat Clca4, from pregnant rat uterus. The gene shares approximately 83% and 70% nucleotide homology with mouse Clca6 and human CLCA4, respectively, and was expressed primarily in rat uterus. The transcripts were upregulated at Gestational Day 22 (prior to parturition), implying a functional involvement in parturition. Western blot analysis showed that rat CLCA4 protein was present in uterus, lung, and heart, but not in any other tissues examined. Confocal microscopy revealed that rat CLCA4 is localized in cell membrane and could not be removed by alkaline or PBS washing. Transient transfection of rat CLCA4-enhanced green fluorescent protein in Chinese hamster ovary cells resulted in production of characteristic Cl(-) currents that could be activated by Ca(2+) and ionomycin but inhibited by niflumic acid, a CLCA-channel blocker. The identification and characterization of rat Clca4 help decipher the contribution of Ca(2+)-activated Cl(-) conductance in myometrial contractility.