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Testis determination requires insulin receptor family function in mice


In mice, gonads are formed shortly before embryonic day 10.5 by the thickening of the mesonephros and consist of somatic cells and migratory primordial germ cells1. The male sex-determining process is set in motion by the sex-determining region of the Y chromosome (Sry), which triggers differentiation of the Sertoli cell lineage. In turn, Sertoli cells function as organizing centres and direct differentiation of the testis. In the absence of Sry expression, neither XX nor XY gonads develop testes2, and alterations in Sry expression are often associated with abnormal sexual differentiation3,4,5,6,7,8. The molecular signalling mechanisms by which Sry specifies the male pathway and models the undifferentiated gonad are unknown. Here we show that the insulin receptor tyrosine kinase family, comprising Ir, Igf1r and Irr, is required for the appearance of male gonads and thus for male sexual differentiation. XY mice that are mutant for all three receptors develop ovaries and show a completely female phenotype. Reduced expression of both Sry and the early testis-specific marker Sox9 indicates that the insulin signalling pathway is required for male sex determination.

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We thank R. Menon, Y.-J. Li, S. McKinnon, Z. Jorai, J. Kargel,T. Shipman, J.-L. Pitetti and the genomic platform of the Frontiers in Genetics at the National Center of Competence in Research for technical assistance; all members of the Parada laboratory for discussions; A. McMahon, R. Behringer, K. Parker, R. Lovell-Badge and J. Dean for probes; and J. Wilson, K. Parker, J. Graff, J. Goldstein and G. Karsenty for critically reading the manuscript. This work was funded by an Excellence in Education Endowment to L.F.P.

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Correspondence to Luis F. Parada.

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The authors declare that they have no competing financial interests.

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Supplementary Figure 1 (JPG 76 kb)

Supplementary Figure 2a (JPG 27 kb)

Supplementary Figure 2b (JPG 31 kb)

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Supplementary Figure 4 part 1 (JPG 96 kb)

Supplementary Figure 4 part 2 (JPG 145 kb)

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Supplementary Figure and Table Legends (DOC 258 kb)

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Figure 1: Mice lacking Ir, Irr and Igf1r show complete gonadal sex reversal at the histological level.
Figure 2: Mice lacking Ir, Irr and Igf1r show complete gonadal sex reversal at the molecular level.
Figure 3: Insulin family signalling is required for early male gonad differentiation.
Figure 4: Male-to-female sex reversal phenotype is variable in Ir Igf1r double mutant and in compound heterozygous Ir+/-Igf1r-/-Irr-/- mutant mice.


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