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: 71/2/613    most recent biolreprod.103.025635v1 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 PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Kirby, J. L. Articles by Hinton, B. T. Search for Related Content PubMed PubMed Citation Articles by Kirby, J. L. Articles by Hinton, B. T. Agricola Articles by Kirby, J. L. Articles by Hinton, B. T.

Characterization of Epididymal Epithelial Cell-Specific Gene Promoters by In Vivo Electroporation¹

Department of Cell Biology³ Department of Internal Medicine,⁴ University of Virginia Health System, Charlottesville, Virginia 22908 Department of Cell Biology and Biochemistry,⁵ Texas Tech University Health Sciences Center, Lubbock, Texas 79430

The mammalian epididymis plays a critical role in sperm maturation, a function dependent on testicular androgens. However, the function of the initial segment, the most proximal part of the epididymis, is also dependent on luminal factors of testicular origin. Efferent duct ligation (EDL), which prevents luminal testicular fluid from reaching the epididymis, results in changes in gene expression within this region. Cystatin-related epididymal spermatogenic (cres) gene and -glutamyl transpeptidase (GGT) mRNA IV are highly expressed in the initial segment and are regulated by luminal testicular factors. EDL results in decreased expression of both genes. To evaluate these promoters in the context of their native physiological state, an in vivo electroporation procedure was used. Significant differences were observed in vivo compared to previous in vitro results. Whereas two C/EBP sites were necessary for transcriptional activity from a 135-base-pair (bp) cres promoter in vitro, only the 5' site displayed functional activity in the in vivo system. A 135-bp GGT promoter IV construct was sufficient for reporter gene expression in vitro. However, in vivo, substantial expression was not observed until the construct was extended to 530 bp. Three polyoma enhancer activator 3 (PEA3) sites were found to be necessary for in vivo reporter gene expression from this construct. A cis-acting negative regulatory element between –530 and –681 bp was also identified that was not previously recognized in the in vitro studies. These studies demonstrate the utility of in vivo electroporation for elucidating promoter elements that may not be identified when traditional in vitro methods are used.

¹ Supported by NIH grants HD32979 to B.T.H. and HD33903 to G.A.C. J.L.K. is supported by a grant from the Medical Scientist Training Program (NIH grant 2T32 GM07267).

² Correspondence: Barry T. Hinton, P.O. Box 800732, University of Virginia Health System, Charlottesville, VA 22908. FAX: 434 982 3912; bth7c{at}virginia.edu

This article has been cited by other articles:

				W. R. Lagor, R. Heller, E. D. De Groh, and G. C. Ness Functional Analysis of the Hepatic HMG-CoA Reductase Promoter by In Vivo Electroporation Experimental Biology and Medicine, March 1, 2007; 232(3): 353 - 361. [Abstract] [Full Text] [PDF]

				L. Yang, S. A. Fox, J. L. Kirby, B. V. Troan, and B. T. Hinton Putative Regulation of Expression of Members of the Ets Variant 4 Transcription Factor Family and Their Downstream Targets in the Rat Epididymis Biol Reprod, April 1, 2006; 74(4): 714 - 720. [Abstract] [Full Text] [PDF]

				E. V. Zubkova and B. Robaire Editorial Commentary J Androl, September 1, 2005; 26(5): 638 - 640. [Full Text] [PDF]