title: Mullerian inhibiting substance is required for germ cell proliferation during early gonadal differentiation in medaka (Oryzias latipes).
abstract: Müllerian inhibiting substance (MIS) is a glycoprotein belonging to the TGF-beta superfamily. In mammals, MIS is responsible for the regression of Müllerian ducts in the male fetus. However, the role of MIS in gonadal sex differentiation of teleost fish, which have no Müllerian ducts, has yet to be clarified. In the present study, we examined the expression pattern of mis and mis type 2 receptor (misr2) mRNAs and the function of MIS signaling in early gonadal differentiation in medaka (teleost, Oryzias latipes). In situ hybridization showed that both mis and misr2 mRNAs were expressed in the somatic cells surrounding the germ cells of both sexes during early sex differentiation. Loss-of-function of either MIS or MIS type II receptor (MISRII) in medaka resulted in suppression of germ cell proliferation during sex differentiation. These results were supported by cell proliferation assay using 5-bromo-2'-deoxyuridine labeling analysis. Treatment of tissue fragments containing germ cells with recombinant eel MIS significantly induced germ cell proliferation in both sexes compared with the untreated control. On the other hand, culture of tissue fragments from the MIS- or MISRII-defective embryos inhibited proliferation of germ cells in both sexes. Moreover, treatment with recombinant eel MIS in the MIS-defective embryos dose-dependently increased germ cell number in both sexes, whereas in the MISRII-defective embryos, it did not permit proliferation of germ cells. These results suggest that in medaka, MIS indirectly stimulates germ cell proliferation through MISRII, expressed in the somatic cells immediately after they reach the gonadal primordium.
authors.affiliation: Graduate School of Science and Technology, Kumamoto University, 2-39-1 Kurokami, Kumamoto, Japan.
authors.forename: Eri
authors.initials: E
authors.lastname: Shiraishi
authors.forename: Norifumi
authors.initials: N
authors.lastname: Yoshinaga
authors.forename: Takeshi
authors.initials: T
authors.lastname: Miura
authors.forename: Hayato
authors.initials: H
authors.lastname: Yokoi
authors.forename: Yuko
authors.initials: Y
authors.lastname: Wakamatsu
authors.forename: Shin-Ichi
authors.initials: S
authors.lastname: Abe
authors.forename: Takeshi
authors.initials: T
authors.lastname: Kitano
mesh_terms: D000818:Animals; D054304:Anti-Mullerian Hormone / Q000502:physiology*; D049109:Cell Proliferation; D005260:Female; D005854:Germ Cells / Q000166:cytology*; D008297:Male; D009990:Oryzias; D018000:Receptors, Peptide / Q000502:physiology*; D018125:Receptors, Transforming Growth Factor beta / Q000502:physiology*; D011994:Recombinant Proteins / Q000494:pharmacology; D012733:Sex Differentiation
chemical_list: D018000:Receptors, Peptide; D018125:Receptors, Transforming Growth Factor beta; D011994:Recombinant Proteins; C084621:anti-Mullerian hormone receptor; D054304:Anti-Mullerian Hormone