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.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

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.


  1. 1

    Capel, B. The battle of the sexes. Mech. Dev. 92, 89–103 (2000)

    CAS  Article  Google Scholar 

  2. 2

    Capel, B. et al. Deletion of Y chromosome sequences located outside the testis determining region can cause XY female sex reversal. Nature Genet. 5, 301–307 (1993)

    CAS  Article  Google Scholar 

  3. 3

    Lee, C. H. & Taketo, T. Normal onset, but prolonged expression, of Sry gene in the B6.YDOM sex-reversed mouse gonad. Dev. Biol. 165, 442–452 (1994)

    CAS  Article  Google Scholar 

  4. 4

    Washburn, L. L., Albrecht, K. H. & Eicher, E. M. C57BL/6J-T-associated sex reversal in mice is caused by reduced expression of a Mus domesticus Sry allele. Genetics 158, 1675–1681 (2001)

    CAS  PubMed  PubMed Central  Google Scholar 

  5. 5

    Tevosian, S. G. et al. Gonadal differentiation, sex determination and normal Sry expression in mice require direct interaction between transcription partners GATA4 and FOG2. Development 129, 4627–4634 (2002)

    CAS  PubMed  Google Scholar 

  6. 6

    Nagamine, C. M., Morohashi, K., Carlisle, C. & Chang, D. K. Sex reversal caused by Mus musculus domesticus Y chromosomes linked to variant expression of the testis-determining gene. Sry. Dev. Biol. 216, 182–194 (1999)

    CAS  Article  Google Scholar 

  7. 7

    Hammes, A. et al. Two splice variants of the Wilms' tumor 1 gene have distinct functions during sex determination and nephron formation. Cell 106, 319–329 (2001)

    CAS  Article  Google Scholar 

  8. 8

    Albrecht, K. H., Young, M., Washburn, L. L. & Eicher, E. M. Sry expression level and protein isoform differences play a role in abnormal testis development in C57BL/6J mice carrying certain Sry alleles. Genetics 164, 277–288 (2003)

    CAS  PubMed  PubMed Central  Google Scholar 

  9. 9

    Accili, D. et al. Early neonatal death in mice homozygous for a null allele of the insulin receptor gene. Nature Genet. 12, 106–109 (1996)

    CAS  Article  Google Scholar 

  10. 10

    Joshi, R. L. et al. Targeted disruption of the insulin receptor gene in the mouse results in neonatal lethality. EMBO J. 15, 1542–1547 (1996)

    CAS  Article  Google Scholar 

  11. 11

    Liu, J. P., Baker, J., Perkins, A. S., Robertson, E. J. & Efstratiadis, A. Mice carrying null mutations of the genes encoding insulin-like growth factor I (Igf-1) and type 1 IGF receptor (Igf1r). Cell 75, 59–72 (1993)

    CAS  Google Scholar 

  12. 12

    Louvi, A., Accili, D. & Efstratiadis, A. Growth-promoting interaction of IGF-II with the insulin receptor during mouse embryonic development. Dev. Biol. 189, 33–48 (1997)

    CAS  Article  Google Scholar 

  13. 13

    Kitamura, T. et al. Preserved pancreatic β-cell development and function in mice lacking the insulin receptor-related receptor. Mol. Cell. Biol. 21, 5624–5630 (2001)

    CAS  Article  Google Scholar 

  14. 14

    Schlessinger, J. Cell signaling by receptor tyrosine kinases. Cell 103, 211–225 (2000)

    CAS  Article  Google Scholar 

  15. 15

    Fernandez, A. M. et al. Functional inactivation of the IGF-I and insulin receptors in skeletal muscle causes type 2 diabetes. Genes Dev. 15, 1926–1934 (2001)

    CAS  Article  Google Scholar 

  16. 16

    Nef, S. & Parada, L. F. Cryptorchidism in mice mutant for Insl3. Nature Genet. 22, 295–299 (1999)

    CAS  Article  Google Scholar 

  17. 17

    Swain, A. & Lovell-Badge, R. Mammalian sex determination: a molecular drama. Genes Dev. 13, 755–767 (1999)

    CAS  Article  Google Scholar 

  18. 18

    Vainio, S., Heikkila, M., Kispert, A., Chin, N. & McMahon, A. P. Female development in mammals is regulated by Wnt-4 signalling. Nature 397, 405–409 (1999)

    ADS  CAS  Article  Google Scholar 

  19. 19

    Soyal, S. M., Amleh, A. & Dean, J. FIGα, a germ cell-specific transcription factor required for ovarian follicle formation. Development 127, 4645–4654 (2000)

    CAS  PubMed  Google Scholar 

  20. 20

    Parr, B. A. & McMahon, A. P. Sexually dimorphic development of the mammalian reproductive tract requires Wnt-7a. Nature 395, 707–710 (1998)

    ADS  CAS  Article  Google Scholar 

  21. 21

    Clarkson, M. J. & Harley, V. R. Sex with two SOX on: SRY and SOX9 in testis development. Trends Endocrinol. Metab. 13, 106–111 (2002)

    CAS  Article  Google Scholar 

  22. 22

    Schmahl, J. & Capel, B. Cell proliferation is necessary for the determination of male fate in the gonad. Dev. Biol. 258, 264–276 (2003)

    CAS  Article  Google Scholar 

  23. 23

    Burgoyne, P. & Palmer, S. in Gonadal Development and Function (ed. Hiller, S. G.) 17–29 (Raven, New York, 1993)

    Google Scholar 

  24. 24

    Colvin, J. S., Green, R. P., Schmahl, J., Capel, B. & Ornitz, D. M. Male-to-female sex reversal in mice lacking fibroblast growth factor 9. Cell 104, 875–889 (2001)

    CAS  Article  Google Scholar 

  25. 25

    Scherer, G. Introduction: vertebrate sex determination and gonadal differentiation. Cell. Mol. Life Sci. 55, 991–909 (1999)

    Article  Google Scholar 

  26. 26

    Jeske, Y. W., Bowles, J., Greenfield, A. & Koopman, P. Expression of a linear Sry transcript in the mouse genital ridge. Nature Genet. 10, 480–482 (1995)

    CAS  Article  Google Scholar 

  27. 27

    Kernie, S. G., Erwin, T. M. & Parada, L. F. Brain remodeling due to neuronal and astrocytic proliferation after controlled cortical injury in mice. J. Neurosci Res. 66, 317–326 (2001)

    CAS  Article  Google Scholar 

Download references


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.

Author information



Corresponding author

Correspondence to Luis F. Parada.

Ethics declarations

Competing interests

The authors declare that they have no competing financial interests.

Supplementary information

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Nef, S., Verma-Kurvari, S., Merenmies, J. et al. Testis determination requires insulin receptor family function in mice. Nature 426, 291–295 (2003). https://doi.org/10.1038/nature02059

Download citation

Further reading


By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.


Sign up for the Nature Briefing newsletter for a daily update on COVID-19 science.
Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing