Cloning and characterization of a calcium-activated chloride channel in rat uterus.
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.
Song
Junfang
J
Institute of Genetics and Developmental Biology and State Key Laboratory of Brain and Cognitive Science, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China.
Zhang
Xiaojuan
X
Qi
Zhi
Z
Sun
Gang
G
Chi
Shaopeng
S
Zhu
Zhaohui
Z
Ren
Jiangong
J
Qiu
Zhaojun
Z
Liu
Ka
K
Myatt
Leslie
L
Ma
Runlin Z
RZ
D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D018118:Chloride Channels / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D003001:Cloning, Molecular; D006224:Cricetinae; D003412:Cricetulus; D005260:Female; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D008969:Molecular Sequence Data; D009215:Myometrium / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D011247:Pregnancy; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017386:Sequence Homology, Amino Acid; D014590:Uterine Contraction / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D014599:Uterus / Q000378:metabolism*
D018118:Chloride Channels; C541949:Clca4 protein, rat; D012333:RNA, Messenger