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BOR - Papers in Press, published online ahead of print January 19, 2005.
Biol Reprod 2005, 10.1095/biolreprod.104.037689
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BIOLOGY OF REPRODUCTION 72, 1151–1160 (2005)
DOI: 10.1095/biolreprod.104.037689
© 2005 by the Society for the Study of Reproduction, Inc.

Androgen Regulation of Stage-Dependent Cyclin D2 Expression in Sertoli Cells Suggests a Role in Modulating Androgen Action on Spermatogenesis1

K.A.L. Tan 4, K.J. Turner 3 4, P.T.K. Saunders 4, G. Verhoeven 5, K. De Gendt 5, N. Atanassova 6, and R.M. Sharpe 2 4

MRC Human Reproductive Sciences Unit,4 Centre for Reproductive Biology, University of Edinburgh, Edinburgh EH16 4SB, Scotland, United Kingdom Laboratory for Experimental Medicine and Endocrinology,5 Department of Developmental Biology, Catholic University of Leuven, B-3000 Leuven, Belgium Institute of Experimental Morphology and Anthropology,6 Bulgarian Academy of Science, 1113 Sofia, Bulgaria

Regulation of spermatogenesis involves stage-dependent androgen action on Sertoli cells, but the pathways involved are unclear. We assessed if cyclin D2 could play a role. In rats, Sertoli cell nuclear, stage-dependent immunoexpression of cyclin D2 switched on after Day 10 and persisted through Day 35, but disappeared by adulthood. However, ethane dimethane sulfonate (EDS)-induced testosterone withdrawal in adult rats for 6 days induced stage-dependent cyclin D2 immunoexpression in Sertoli cells, with highest expression at stages IX-XII and nondetectable at stages VI–VIII (opposite that for androgen receptor [AR] immunoexpression). In EDS-treated rats, a single injection of testosterone but not of estrogen reversed this change in 4 h, and testosterone administration from the time of EDS treatment prevented expression of cyclin D2 in Sertoli cells. The EDS-induced changes in cyclin D2 immunoexpression were matched by changes in expression of Ccnd2 (cyclin D2) mRNA in isolated stage-dissected tubules. Treatment of adult rats with flutamide induced stage-dependent cyclin D2 immunoexpression in Sertoli cells within 18 h, and confocal microscopy revealed that immunoexpression of AR and cyclin D2 were mutually exclusive within individual seminiferous tubules in these animals. Sertoli cell-selective ablation of the AR in mice using Cre/loxP technology also resulted in stage-dependent Sertoli cell cyclin D2 immunoexpression. Downstream from cyclin D2 action is retinoblastoma 1 (RB1), a tumor suppressor protein, immunoexpression of which paralleled stage-dependent AR expression in Sertoli cells; RB1 stage specificity disappeared after EDS treatment. These results point to a non-cell cycle role for cyclin D2 and RB1 in mature Sertoli cells in the stage-dependent mechanisms regulated by AR expression and androgen action.

Sertoli cells, spermatogenesis, testosterone

1 Supported by MRC, Wellcome Trust, Concerted Research Action (Research Fund Catholic University of Leuven) and Fund for Scientific Research-Flanders (Belgium).

2 Correspondence. FAX: 44 131 242 6231; r.sharpe{at}hrsu.mrc.ac.uk

3 Current address: MRL Department of Safety Assessment, Merck & Co Inc, WP45-120 West Point, PA 19486




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