title: In vitro androgen-induced growth and morphogenesis of the Wolffian duct within urogenital ridge.
abstract: Male mouse urogenital ridges (URs) at 15.5 days of gestation (vaginal plug = day 0) containing Wolffian (WDs) and Mullerian ducts were cultured for 4 days with or without gonads in serum-free medium (1:1 mixture of Dulbecco's modified Eagle's medium and Ham's F-12 supplemented with insulin, transferrin, cholera toxin, epidermal growth factor, and BSA). URs without gonads were grown in serum-free medium with testosterone (T, 10(-7) M), 5 alpha-dihydrotestosterone (DHT, 10(-8) M), T (10(-7) M) plus cyproterone acetate (antiandrogen, 10(-5) M), or T (10(-8) M) plus 390 MSD (17 beta-N, N-diisopropylcarbamoyl-4-aza-5 alpha- androstan-3-one, an inhibitor of 5 alpha-reductase, 10(-5) M). After 4 days of culture the number of epididymal curvatures that appeared in the upper portion of WDs were quantified. DNA content of URs grown in serum-free medium was also measured. Both T and DHT increased DNA contents in a dose- (T = 10(-8) to 10(-10) M, DHT = 10(-9) to 10(-11) M) and time-dependent manner. DHT was approximately 10-fold more effective than T in eliciting epididymal coiling and increasing DNA content of URs. The effects of T or DHT were mimicked by coculture with fetal testes. Epididymal coiling and an increase in DNA content occurred in URs grown in the presence of T plus 390 MSD. By contrast, URs cultured without androgens or with T plus cyproterone acetate failed to undergo epididymal coiling and to increase DNA content. The conversion rate per mg protein of [1 beta, 2 beta-3H]T into [3H]DHT was 0.30-fold lower in 15.5 day URs cultured over a 4-day period in comparison to urogenital sinuses whose development is known to be dependent upon DHT. These data suggest that T is an important hormone in the development of the upper portion of WDs, although it is not possible to exclude a role for DHT in the development of the epididymis.
authors.affiliation: Department of Anatomy, University of California, San Francisco 94143.
authors.forename: M
authors.initials: M
authors.lastname: Tsuji
authors.forename: H
authors.initials: H
authors.lastname: Shima
authors.forename: G R
authors.initials: GR
authors.lastname: Cunha
mesh_terms: D000728:Androgens / Q000494:pharmacology*; D000818:Animals; D002772:Cholera Toxin / Q000494:pharmacology; D003534:Cyproterone / Q000031:analogs & derivatives / Q000494:pharmacology; D017373:Cyproterone Acetate; D004247:DNA / Q000378:metabolism; D013196:Dihydrotestosterone / Q000494:pharmacology; D004815:Epidermal Growth Factor / Q000494:pharmacology; D004822:Epididymis / Q000196:embryology; D004848:Epithelium / Q000254:growth & development; D005865:Gestational Age; D007328:Insulin / Q000494:pharmacology; D008297:Male; D008648:Mesoderm / Q000502:physiology; D051379:Mice; D008807:Mice, Inbred BALB C; D009024:Morphogenesis / Q000187:drug effects*; D009924:Organ Culture Techniques; D013739:Testosterone / Q000494:pharmacology; D014168:Transferrin / Q000494:pharmacology; D014928:Wolffian Ducts / Q000187:drug effects / Q000254:growth & development*
chemical_list: D000728:Androgens; D007328:Insulin; D014168:Transferrin; D013196:Dihydrotestosterone; D013739:Testosterone; D017373:Cyproterone Acetate; D004815:Epidermal Growth Factor; D004247:DNA; D002772:Cholera Toxin; D003534:Cyproterone