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Endocrinology Vol. 140, No. 5 2343-2352
Copyright © 1999 by The Endocrine Society

ARTICLES

Action of Retinoids on Embryonic and Early Postnatal Testis Development

Andrea S. Cupp, Jannette M. Dufour, Grace Kim, Michael K. Skinner and Kwan Hee Kim

Center for Reproductive Biology, Department of Genetics and Cell Biology and Department of Biochemistry and Biophysics, Washington State University, Pullman, Washington 99164-4231

Address all correspondence and requests for reprints to: Michael K. Skinner, Center for Reproductive Biology, Department of Genetics and Cell Biology, Washington State University, Pullman, Washington 99164-4231. E-mail: skinner{at}mail.wsu.edu

The current study investigates the hypothesis that retinoids have a role in embryonic testis development. The action of retinoids on testis development and the expression of retinoic acid receptors (RAR{alpha}, RARß, RAR{gamma}) were examined. In embryonic day 13 (E13; plug date = E0) testis organ cultures an RAR-selective agonist and all-trans retinoic acid completely inhibited seminiferous cord formation. In contrast, an RAR{alpha}-selective antagonist had no effect. RT-PCR demonstrated that RAR{alpha} messenger RNA (mRNA) was expressed at all developmental time points evaluated, which included embryonic day 14 (E14) through postnatal day 30 (P30). Expression of RARß mRNA was present at E15 through P2, whereas RAR{gamma} mRNA was expressed at E18 through P2. Cellular localization of receptors by immunohistochemistry indicated that RAR{alpha} was localized to the interstitium at E18 and to the seminiferous cords by P0. RARß and RAR{gamma} were detected in both interstitium and cords at E16 and by E18 were mainly expressed in the cords. At P0 RARß and RAR{gamma} were localized to the germ cell populations. To examine retinoid actions, the growth of P0 testis cultures were investigated. Interestingly, retinol and retinoic acid did not inhibit growth of P0 testis cultures but did inhibit the action of growth stimulators. Retinoic acid inhibited FSH, EGF, and 10% calf serum stimulated growth in P0 testis cultures. The hypothesis tested was that the inhibitory effects of retinoids on P0 testis growth may be mediated through the growth inhibitor, transforming growth factor-ß (TGFß). The action of retinoids on TGFß mRNA expression was examined in P0 testis cultures. Retinoic acid stimulated TGFß3 mRNA expression within 24 h and increased expression of TGFß1 and TGFß2 after 72 h. Retinol increased expression of TGFß1 and TGFß2 but not TGFß3 after 72 h of treatment. These observations indicate that retinoic acid can influence seminiferous cord formation and testis growth. The inhibitory actions of retinoids may in part be mediated through increased expression of TGFß isoforms.




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