Abstract
RSPO1 is a newly discovered gene involved in sex differentiation. Two goat BAC clones encompassing the RSPO1 gene (gRSPO1) were injected into mouse oocytes and several transgenic lines derived. Both clones induced gRSPO1 over-expression in various tissues, including male and female gonads, with no obvious phenotype and normal sex-ratios. Introgression of the gRSPO1 transgene into a mouse RSPO1 knockout genotype resulted in the rescue of the fertility and the disappearance of the masculinized gonadic features of the females, demonstrating the functionality of the goat protein in a mouse context. On the contrary, over-expression of gRSPO1 within a mSRY or a gSRY-XX genotypes did not interfere with the SRY-induced male phenotype.
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References
Bernard P, Harley VR (2007) Wnt4 action in gonadal development and sex determination. Int J Biochem Cell Biol 39:31–43. doi:10.1016/j.biocel.2006.06.007
Bernard P, Sim H, Knower K, Vilain E, Harley V (2008) Human SRY inhibits beta-catenin-mediated transcription. Int J Biochem Cell Biol 40:2889–2900. doi:10.1016/j.biocel.2008.06.006
Binnerts ME, Kim KA, Bright JM, Patel SM, Tran K, Zhou M, Leung JM, Liu Y, Lomas WE 3rd, Dixon M, Hazell SA, Wagle M, Nie WS, Tomasevic N, Williams J, Zhan X, Levy MD, Funk WD, Abo A (2007) R-Spondin1 regulates Wnt signaling by inhibiting internalization of LRP6. Proc Natl Acad Sci USA 104:14700–14705. doi:10.1073/pnas.0702305104
Brennan J, Capel B (2004) One tissue, two fates: molecular genetic events that underlie testis versus ovary development. Nat Rev Genet 5:509–521. doi:10.1038/nrg1381
Chassot AA, Ranc F, Gregoire EP, Roepers-Gajadien HL, Taketo MM, Camerino G, de Rooij DG, Schedl A, Chaboissier MC (2008) Activation of beta-catenin signaling by Rspo1 controls differentiation of the mammalian ovary. Hum Mol Genet 17:1264–1277. doi:10.1093/hmg/ddn016
Jeays-Ward K, Hoyle C, Brennan J, Dandonneau M, Alldus G, Capel B, Swain A (2003) Endothelial and steroidogenic cell migration are regulated by WNT4 in the developing mammalian gonad. Development 130:3663–3670. doi:10.1242/dev.00591
Jordan BK, Mohammed M, Ching ST, Delot E, Chen XN, Dewing P, Swain A, Rao PN, Elejalde BR, Vilain E (2001) Up-regulation of WNT-4 signaling and dosage-sensitive sex reversal in humans. Am J Hum Genet 68:1102–1109. doi:10.1086/320125
Kim KA, Kakitani M, Zhao J, Oshima T, Tang T, Binnerts M, Liu Y, Boyle B, Park E, Emtage P, Funk WD, Tomizuka K (2005) Mitogenic influence of human R-spondin1 on the intestinal epithelium. Science 309:1256–1259. doi:10.1126/science.1112521
Kim Y, Kobayashi A, Sekido R, DiNapoli L, Brennan J, Chaboissier MC, Poulat F, Behringer RR, Lovell-Badge R, Capel B (2006) Fgf9 and Wnt4 act as antagonistic signals to regulate mammalian sex determination. PLoS Biol 4:e187. doi:10.1371/journal.pbio.0040187
Kim KA, Wagle M, Tran K, Zhan X, Dixon MA, Liu S, Gros D, Korver W, Yonkovich S, Tomasevic N, Binnerts M, Abo A (2008) R-Spondin family members regulate the Wnt pathway by a common mechanism. Mol Biol Cell 19:2588–2596. doi:10.1091/mbc.E08-02-0187
Kocer A, Pinheiro I, Pannetier M, Renault L, Parma P, Radi O, Kim KA, Camerino G, Pailhoux E (2008) R-spondin1 and FOXL2 act into two distinct cellular types during goat ovarian differentiation. BMC Dev Biol 8:36. doi:10.1186/1471-213X-8-36
Koopman P, Gubbay J, Vivian N, Goodfellow P, Lovell-Badge R (1991) Male development of chromosomally female mice transgenic for Sry. Nature 351:117–121. doi:10.1038/351117a0
Maatouk DM, DiNapoli L, Alvers A, Parker KL, Taketo MM, Capel B (2008) Stabilization of beta-catenin in XY gonads causes male-to-female sex-reversal. Hum Mol Genet 17:2949–2955. doi:10.1093/hmg/ddn193
Mizusaki H, Kawabe K, Mukai T, Ariyoshi E, Kasahara M, Yoshioka H, Swain A, Morohashi K (2003) Dax-1 (dosage-sensitive sex reversal-adrenal hypoplasia congenita critical region on the X chromosome, gene 1) gene transcription is regulated by wnt4 in the female developing gonad. Mol Endocrinol 17:507–519. doi:10.1210/me.2002-0362
Nam JS, Turcotte TJ, Yoon JK (2007) Dynamic expression of R-spondin family genes in mouse development. Gene Expr Patterns 7:306–312. doi:10.1016/j.modgep.2006.08.006
Ottolenghi C, Pelosi E, Tran J, Colombino M, Douglass E, Nedorezov T, Cao A, Forabosco A, Schlessinger D (2007) Loss of Wnt4 and Foxl2 leads to female-to-male sex reversal extending to germ cells. Hum Mol Genet 16:2795–2804. doi:10.1093/hmg/ddm235
Pailhoux E, Vigier B, Chaffaux S, Servel N, Taourit S, Furet JP, Fellous M, Grosclaude F, Cribiu EP, Cotinot C, Vaiman D (2001) A 11.7-kb deletion triggers intersexuality and polledness in goats. Nat Genet 29:453–458. doi:10.1038/ng769
Pannetier M, Tilly G, Kocer A, Hudrisier M, Renault L, Chesnais N, Costa J, Le Provost F, Vaiman D, Vilotte JL, Pailhoux E (2006) Goat SRY induces testis development in XX transgenic mice. FEBS Lett 580:3715–3720. doi:10.1016/j.febslet.2006.05.060
Parma P, Radi O, Vidal V, Chaboissier MC, Dellambra E, Valentini S, Guerra L, Schedl A, Camerino G (2006) R-spondin1 is essential in sex determination, skin differentiation and malignancy. Nat Genet 38:1304–1309. doi:10.1038/ng1907
Schedl A, Larin Z, Montoliu L, Thies E, Kelsey G, Lehrach H, Schutz G (1993) A method for the generation of YAC transgenic mice by pronuclear microinjection. Nucleic Acids Res 21:4783–4787. doi:10.1093/nar/21.20.4783
Schibler L, Vaiman D, Oustry A, Guinec N, Dangy-Caye AL, Billault A, Cribiu EP (1998) Construction and extensive characterization of a goat bacterial artificial chromosome library with threefold genome coverage. Mamm Genome 9:119–124. doi:10.1007/s003359900701
Smith CA, Shoemaker CM, Roeszler KN, Queen J, Crews D, Sinclair AH (2008) Cloning and expression of R-Spondin1 in different vertebrates suggests a conserved role in ovarian development. BMC Dev Biol 8:72. doi:10.1186/1471-213X-8-72
Tomizuka K, Horikoshi K, Kitada R, Sugawara Y, Iba Y, Kojima A, Yoshitome A, Yamawaki K, Amagai M, Inoue A, Oshima T, Kakitani M (2008) R-spondin1 plays an essential role in ovarian development through positively regulating Wnt-4 signaling. Hum Mol Genet 17:1278–1291. doi:10.1093/hmg/ddn036
Uda M, Ottolenghi C, Crisponi L, Garcia JE, Deiana M, Kimber W, Forabosco A, Cao A, Schlessinger D, Pilia G (2004) Foxl2 disruption causes mouse ovarian failure by pervasive blockage of follicle development. Hum Mol Genet 13:1171–1181. doi:10.1093/hmg/ddh124
Vainio S, Heikkila M, Kispert A, Chin N, McMahon AP (1999) Female development in mammals is regulated by Wnt-4 signalling. Nature 397:405–409. doi:10.1038/17068
Wei Q, Yokota C, Semenov MV, Doble B, Woodgett J, He X (2007) R-spondin1 is a high affinity ligand for LRP6 and induces LRP6 phosphorylation and beta-catenin signaling. J Biol Chem 282:15903–15911. doi:10.1074/jbc.M701927200
Whitelaw CB, Springbett AJ, Webster J, Clark J (1993) The majority of G0 transgenic mice are derived from mosaic embryos. Transgenic Res 2:29–32. doi:10.1007/BF01977678
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This work was partially supported by the ANR-06, GenAnimal TEGOD.
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Laurine Buscara and Fatemeh Montazer-Torbati contributed equally to this work.
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Buscara, L., Montazer-Torbati, F., Chadi, S. et al. Goat RSPO1 over-expression rescues sex-reversal in Rspo1-knockout XX mice but does not perturb testis differentiation in XY or sex-reversed XX mice. Transgenic Res 18, 649–654 (2009). https://doi.org/10.1007/s11248-009-9247-2
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DOI: https://doi.org/10.1007/s11248-009-9247-2