Kit/stem cell factor receptor-induced activation of phosphatidylinositol 3′-kinase is essential for male fertility

Abstract

The c-kit-encoded transmembrane tyrosine kinase receptor for stem cell factor (Kit/SCF-R) is required for normal haematopoiesis, melanogenesis and gametogenesis1,2,3. However, the roles of individual Kit/SCF-R-induced signalling pathways in the control of developmental processes in the intact animal are completely unknown. To examine the function of SCF-induced phosphatidylinositol (PI) 3′-kinase activation in vivo, we employed the Cre-loxP system4 to mutate the codon for Tyr719, the PI 3′-kinase binding site in Kit/SCF-R, to Phe in the genome of mice by homologous recombination. Homozygous (Y719F/Y719F) mutant mice are viable. The mutation completely disrupted PI 3′-kinase binding to Kit/SCF-R and reduced SCF-induced PI 3′-kinase-dependent activation of Akt by 90%. The mutation induced a gender- and tissue-specific defect. Although there are no haematopoietic or pigmentation defects in homozygous mutant mice, males are sterile due to a block in spermatogenesis, with initially decreased proliferation and subsequent extensive apoptosis occurring at the spermatogonial stem-cell level. In contrast, female homozygotes are fully fertile. This is the first report so far demonstrating the role of an individual signalling pathway downstream of Kit/SCF-R in the intact animal. It provides the first in vivo model for male sterility caused by a discrete signalling pathway defect affecting early germ cells.

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Figure 1: Disruption of SCF-induced PI 3′-kinase signalling in mice by mutation of the PI 3′-kinase-binding site in Kit/SCF-R.
Figure 2: Testes, ovaries and hair follicles of adult mice.
Figure 3: Primordial germ cells in genital ridges and cell proliferation, apoptosis, and Kit positive cells in parallel sections of testes from P8 pups.
Figure 4: Cell proliferation, apoptosis, and Kit-positive cells in parallel sections of testes from P10 pups.

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Acknowledgements

We thank E.A. Barcarse for help with ES cells; S. Kwoh and B.A. Dominguez for assistance with breeding and blastocyst injections, respectively; E. Saez, Y. Dayn, W. Lin, C. Bentley, S. Crone, J. Morris and J. DeFratis and the staff at the Transgenic Facility at Salk; R. Rottapel for advice on mast cell isolation; D. Lee for advice on total blood cell counts, and C. Barlow for advice on testicular development and histology; T. Chan and I. Gout for antibodies; and N. Carter and M. Latterich for discussion. This work was supported by NIH grants from the NCI to T.H., from HD to K.-F.L., and from GM to S.O'G. P.B.-J. is a Special Fellow of the Leukemia Society of America, K.-F.L. a Pew Scholar, and T.H. a Frank and Else Schilling American Cancer Society Professor.

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Correspondence to Peter Blume-Jensen or Tony Hunter.

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Blume-Jensen, P., Jiang, G., Hyman, R. et al. Kit/stem cell factor receptor-induced activation of phosphatidylinositol 3′-kinase is essential for male fertility. Nat Genet 24, 157–162 (2000) doi:10.1038/72814

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