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BOR - Papers in Press, published online ahead of print September 10, 2008.
Biol Reprod 2008, 10.1095/biolreprod.108.068346
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BIOLOGY OF REPRODUCTION 80, 13–23 (2009)
DOI: 10.1095/biolreprod.108.068346
© 2009 by the Society for the Study of Reproduction, Inc.

Stimulating Effects of Dopamine on Chloride Transport Across the Rat Caudal Epididymal Epithelium in Culture1

Jian-Yang Du 3, Wu-Lin Zuo 3, Ye-Chun Ruan , Zi-Huan Yang , Min-Hui Chen , Si-Liang Chen , Sheng Li , Zhong-Luan Wu , Hui Xiang , and Wen-Liang Zhou  2

School of Life Science, Sun Yat-sen University, Guangzhou 510275, China

ABSTRACT

The present study investigated the effects of dopamine on chloride transport across cultured rat caudal epididymal epithelium. The results showed that dopamine induced a biphasic short-circuit current (Isc) in a concentration-dependent manner. The dopamine-induced response consisted of an initial rapid spike followed by a sustained phase. The alpha and beta adrenoreceptor inhibitors, phentolamine and propranolol, inhibited the initial spike and the sustained phase, respectively, suggesting a contribution of adrenergic receptors. The response was almost abolished by removing the extracellular Cl–, suggesting that the dopamine-induced short-circuit current is primarily a Cl– current. The response was inhibited by the apical Cl– channel blocker, diphenylamine-dicarboxylic acid, and the Ca2+-activated Cl– channel blocker, disulfonic acid stilbene, indicating that Cl– may pass through two types of Cl– channels on the apical side. Preloading monolayers with the intracellular Ca2+ chelator BAPTA/AM abolished the initial spike and greatly reduced the second phase in the Isc response to dopamine. Pretreating the monolayers with an adenylate cyclase inhibitor, MDL12330A, inhibited all of the second Isc response and part of the initial spike. Also, characteristics of the Cl– currents induced by dopamine were observed in whole-cell patch-clamp recording. The increases of intracellular cAMP and Ca2+ induced by dopamine were also measured. The results suggest that extracellular dopamine activates Ca2+-dependent and cAMP-dependent regulatory pathways, leading to activation of both Ca2+-dependent and cAMP-dependent Cl– conductances in epididymal epithelial cells.

calcium, cyclic adenosine monophosphate, dopamine, epididymis, male reproductive tract

FOOTNOTES

1Supported by the National Natural Science Foundation of China (no. 30770817), National 973 project foundation (no. 2006CB504002), and the South China National Research Center for Integrated Biosciences in collaboration with Sun Yat-sen University.

Correspondence: 2FAX: 86 20 84110060; e-mail: lsszwl{at}yahoo.com.cn

3 These authors contributed equally to this work.






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Copyright © 2009 by the Society for the Study of Reproduction.