title: Molecular cloning and characterization of anti-Müllerian hormone (AMH) from the Japanese wrinkled frog, Rana rugosa.
abstract: The role of anti-Müllerian hormone (AMH) during gonad development has been studied extensively in many species of mammal, bird, reptile, and fish but remains unresolved in amphibians. In male mammalian embryos, Sox9 activates AMH expression, which initiates regression of the Müllerian ducts. However, Sox9 (Sry-related HMG box 9) is unlikely to initiate AMH in chicken, because AMH precedes Sox9 expression in this species. To clarify whether AMH is involved in testicular differentiation in amphibians, we cloned the full-length AMH cDNA from the Japanese wrinkled frog, Rana rugosa. The AMH gene, which appears to be autosomal, is exclusively expressed in the testis of adult frog among 8 different tissues examined; Sertoli cells are probably responsible for its expression. AMH expression was found in the undifferentiated gonad of both male and female tadpoles, increasing in the differentiating testis. Moreover, we observed consensus binding sites for Sox9 in the 5'-flanking region of the AMH gene. Sox9 stimulated statistically significant AMH expression in luciferase reporter assays when coexpressed in Xenopus kidney-derived A6 cells. However, Sox9 expression showed no sexual dimorphism when AMH expression was up-regulated in the developing testis. These results, taken together, suggest that AMH is probably involved in testicular differentiation in R. rugosa, although an additional, perhaps tissue-specific, transcription factor may be required for the regulation of AMH transcription.
authors.affiliation: Department of Biology (M.K., M.S., D.S., T.I., Y.Matsuo, S.M., M.N.), Faculty of Education and Integrated Arts and Sciences, Waseda University, Shinjuku-ku, Tokyo 162-8480, Japan; Laboratory of Animal Genetics (Y.U., Y.Matsud.), Department of Applied Molecular Biosciences, Graduate School of Bioagricultural Science, Nagoya University, Chikusa-ku, Nagoya, 464-8601, Japan; Department of Science Education (Y.N.), Faculty of Education, Ehime University, Matsuyama, Ehime 790-8577, Japan; and Laboratory of Reproductive and Developmental Biology (Y.K.), Faculty of Science, Hokkaido University, Sapporo 060-0810, Japan.
authors.forename: Maho
authors.initials: M
authors.lastname: Kodama
authors.forename: Mari
authors.initials: M
authors.lastname: Suda
authors.forename: Daiki
authors.initials: D
authors.lastname: Sakamoto
authors.forename: Takehiro
authors.initials: T
authors.lastname: Iwasaki
authors.forename: Yasuki
authors.initials: Y
authors.lastname: Matsuo
authors.forename: Yoshinobu
authors.initials: Y
authors.lastname: Uno
authors.forename: Yoichi
authors.initials: Y
authors.lastname: Matsuda
authors.forename: Yoriko
authors.initials: Y
authors.lastname: Nakamura
authors.forename: Shun
authors.initials: S
authors.lastname: Maekawa
authors.forename: Yoshinao
authors.initials: Y
authors.lastname: Katsu
authors.forename: Masahisa
authors.initials: M
authors.lastname: Nakamura
mesh_terms: D000818:Animals; D054304:Anti-Mullerian Hormone / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary; D018507:Gene Expression Regulation, Developmental; D007814:Larva; D008297:Male; D012333:RNA, Messenger / Q000378:metabolism*; D011898:Ranidae; D020133:Reverse Transcriptase Polymerase Chain Reaction; D055758:SOX9 Transcription Factor / Q000235:genetics* / Q000378:metabolism; D013737:Testis / Q000378:metabolism*
chemical_list: D018076:DNA, Complementary; D012333:RNA, Messenger; D055758:SOX9 Transcription Factor; D054304:Anti-Mullerian Hormone