HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: 290/6/C1560    most recent 00468.2005v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Web of Science (6) Citing Articles via Google Scholar Google Scholar Articles by Carlin, R. W. Articles by Schultz, B. D. Search for Related Content PubMed PubMed Citation Articles by Carlin, R. W. Articles by Schultz, B. D.

MEMBRANE TRANSPORTERS, ION CHANNELS, AND PUMPS

PVD9902, a porcine vas deferens epithelial cell line that exhibits neurotransmitter-stimulated anion secretion and expresses numerous HCO₃^– transporters

¹Department of Anatomy and Physiology and ²Department of Animal Science and Industry, Kansas State University, Manhattan, Kansas

Submitted 20 September 2005 ; accepted in final form 6 January 2006

Epithelial ion transport disorders, including cystic fibrosis, adversely affect male reproductive function by nonobstructive mechanisms and by obstruction of the distal duct. Continuous cell lines that could be used to define ion transport mechanisms in this tissue are not readily available. In the present study, porcine vas deferens epithelial cells were isolated by standard techniques, and the cells spontaneously immortalized to form a porcine vas deferens epithelial cell line that we have titled PVD9902. Cells were maintained in continuous culture for >4 yr and 200 passages in a typical growth medium. Frozen stocks were generated, and thawed cells exhibited growth characteristics indistinguishable from their nonfrozen counterparts. Molecular and immunocytochemical studies confirmed the origin and epithelial nature of these cells. When seeded on permeable supports, PVD9902 cells grew as electrically tight (>6,000 ·cm²), confluent monolayers that responded to forskolin with an increase in short-circuit current (I_sc; 8 ± 1 µA/cm²) that required Cl^–, HCO₃^–, and Na⁺, and was partially sensitive to bumetanide. mRNA was expressed for a number of anion transporters, including CFTR, electrogenic Na⁺-HCO₃^– cotransporter 1b (NBCe1b), downregulated in adenoma, pendrin, and Cl^–/formate exchanger. Both forskolin and isoproterenol caused an increase in cellular cAMP levels. In addition, PVD9902 cell monolayers responded to physiological (i.e., adenosine, norepinephrine) and pharmacological [i.e., 5'-(N-ethylcarboxamido)adenosine, isoproterenol] agonists with increases in I_sc. Unlike their freshly isolated counterparts, however, PVD9902 cells did not respond to glucocorticoid exposure with an increase in amiloride-sensitive I_sc. RT-PCR analysis revealed the presence of both glucocorticoid and mineralocorticoid receptor mRNA as well as mRNA for the - and -subunits of the epithelia Na⁺ channels (- and -ENaC), but not -ENaC. Nonetheless, PVD9902 cells recapitulated most observations in freshly isolated cells and thus represent a powerful new tool to characterize mechanisms that contribute to male reproductive function.

male reproductive tract; cystic fibrosis; epithelial Na⁺ channel expression; glucocorticoid receptor; adrenergic; vasopressin

This article has been cited by other articles:

				F. Pierucci-Alves, C. L. Duncan, and B. D. Schultz Testosterone Upregulates Anion Secretion Across Porcine Vas Deferens Epithelia In Vitro Biol Reprod, October 1, 2009; 81(4): 628 - 635. [Abstract] [Full Text] [PDF]

				H. C. Chan, Y. C. Ruan, Q. He, M. H. Chen, H. Chen, W. M. Xu, W. Y. Chen, C. Xie, X. H. Zhang, and Z. Zhou The cystic fibrosis transmembrane conductance regulator in reproductive health and disease J. Physiol., May 15, 2009; 587(10): 2187 - 2195. [Abstract] [Full Text] [PDF]

				F. Pierucci-Alves and B. D. Schultz Bradykinin-Stimulated Cyclooxygenase Activity Stimulates Vas Deferens Epithelial Anion Secretion In Vitro in Swine and Humans Biol Reprod, September 1, 2008; 79(3): 501 - 509. [Abstract] [Full Text] [PDF]

				T. M Hagedorn, R. W Carlin, and B. D Schultz Oxytocin and Vasopressin Stimulate Anion Secretion by Human and Porcine Vas Deferens Epithelia Biol Reprod, September 1, 2007; 77(3): 416 - 424. [Abstract] [Full Text] [PDF]

				K. Nakaya, D. G. Harbidge, P. Wangemann, B. D. Schultz, E. D. Green, S. M. Wall, and D. C. Marcus Lack of pendrin HCO3- transport elevates vestibular endolymphatic [Ca2+] by inhibition of acid-sensitive TRPV5 and TRPV6 channels Am J Physiol Renal Physiol, May 1, 2007; 292(5): F1314 - F1321. [Abstract] [Full Text] [PDF]