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BOR - Papers in Press, published online ahead of print June 7, 2006.
Biol Reprod 2006, 10.1095/biolreprod.106.052316
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biolreprod.106.052316v1
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BIOLOGY OF REPRODUCTION 75, 407–413 (2006)
DOI: 10.1095/biolreprod.106.052316
© 2006 by the Society for the Study of Reproduction, Inc.

Research Article

Cellular Mechanisms of Carbachol-Stimulated Cl– Secretion in Rat Epididymal Epithelium1

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

School of Life Science, Sun Yat-sen University, Guangzhou 510275, People's Republic of China

ABSTRACT

Neurotransmitter-controlled Cl– secretions play an important role in maintenance of the epididymal microenvironment for sperm maturation. This study was carried out to investigate the effect of carbachol (CCH) on the cultured rat epididymal epithelium and the signal transduction mechanisms of this response. In normal K-H solution, CCH added basolaterally elicited a biphasic Isc response consisting of a transient spike followed by a second sustained response. Ca2+ activated Cl– channel blocker disulfonic acid stilbene (DIDS, 300 µM) only inhibited part of the CCH-induced Isc response, while nonselective Cl– channel blocker diphenylamine-dicarboxylic acid (DPC, 1 mM) reduced all, indicating the involvement of different conductance pathways. Both peaks of the CCH-induced Isc response could be significantly inhibited by pretreatment with an adenylate cyclase inhibitor, MDL12330A (50 µM). An increase in intracellular cAMP content upon stimulation of CCH was measured. All of the initial peak and part of the second peak could be inhibited by pretreatment with Ca2+-chelating agent BAPTA/AM (50 µM) and an endoplasmic reticulum Ca2+ pump inhibitor, Thapsigagin (Tg, 1 µM). In a whole-cell patch clamp experiment, CCH induced an inward current in the single cell. Two different profiles of currents were found; the first component current exhibited an outward rectifying I-V relationship in a time and voltage-dependent manner, and the current followed showed a linear I-V relationship. The carbachol-induced current was found to be partially blockable by DIDS and could be completely blocked by DPC. The above results indicate that the CCH-induced Cl– secretion could be mediated by Ca2+ and cAMP-dependent regulatory pathways.

calcium, epididymis, male reproductive tract, neurotransmitters, signal transduction

FOOTNOTES

1 Supported by National Natural Science Foundation of China (contract grant numbers: 30370540 and 30570229).

2 Correspondence: FAX: 86 20 8411 0060; wenliangzhou{at}yahoo.com






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Copyright © 2006 by the Society for the Study of Reproduction.