Abstract
Spermatogonial stem cells (SSCs) produce haploid sperm via mitosis and meiosis in vivo. Although the technique to culture mouse SSCs has been well established, induction of meiosis in vitro has remained a challenge. Retinoic acid (RA) is required for meiosis in vivo; however, RA alone is not sufficient to induce meiosis in vitro. Here, we describe a method in which nutrient restriction and RA synergistically induce meiotic initiation into meiotic prophase I in cultured mouse SSCs.
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References
Kubota H, Brinster RL (2008) Culture of rodent spermatogonial stem cells, male germline stem cells of the postnatal animal. Methods Cell Biol 86:59–84
Hayashi K et al (2011) Reconstitution of the mouse germ cell specification pathway in culture by pluripotent stem cells. Cell 146(4):519–532
Nagano M et al (1998) Culture of mouse spermatogonial stem cells. Tissue Cell 30(4):389–397
Kanatsu-Shinohara M et al (2003) Long-term proliferation in culture and germline transmission of mouse male germline stem cells. Biol Reprod 69:612–616
Kanatsu-Shinohara M et al (2014) Improved serum- and feeder-free culture of mouse germline stem cells. Biol Reprod 91(4):88
Handel MA, Schimenti JC (2010) Genetics of mammalian meiosis: regulation, dynamics and impact on fertility. Nat Rev Genet 11:124–136
Handel MA, Eppig JJ, Schimenti JC (2014) Applying “gold standards” to in-vitro-derived germ cells. Cell 157(6):1257–1261
Hogarth CA, Griswold MD (2010) The key role of vitamin A in spermatogenesis. J Clin Invest 120(4):956–962
Amory JK et al (2011) Suppression of spermatogenesis by bisdichloroacetyldiamines is mediated by inhibition of testicular retinoic acid biosynthesis. J Androl 32(1):111–119
Zhang X, Gunewardena S, Wang N (2021) Nutrient restriction synergizes with retinoic acid to induce mammalian meiotic initiation in vitro. Nat Commun 12(1):1758
Velte EK et al (2019) Differential RA responsiveness directs formation of functionally distinct spermatogonial populations at the initiation of spermatogenesis in the mouse. Development 146(12)
Ferder IC et al (2019) Meiotic gatekeeper STRA8 suppresses autophagy by repressing Nr1d1 expression during spermatogenesis in mice. PLoS Genet 15(5):e1008084
Mizushima N, Levine B (2010) Autophagy in mammalian development and differentiation. Nature Publishing Group, p 12
Efeyan A, Comb WC, Sabatini DM (2015) Nutrient-sensing mechanisms and pathways. Nature 517(7534):302–310
Broach JR (2012) Nutritional control of growth and development in yeast. Genetics 192(1):73–105
Harigaya Y, Yamamoto M (2007) Molecular mechanisms underlying the mitosis-meiosis decision. Chromosome Res 15(5):523–537
Dym M et al (1995) Expression of c-kit receptor and its autophosphorylation in immature rat type A spermatogonia. Biol Reprod 52(1):8–19
Hofmann MC, Braydich-Stolle L, Dym M (2005) Isolation of male germ-line stem cells; influence of GDNF. Dev Biol 279(1):114–124
Wu X et al (2011) Spermatogonial stem cell self-renewal requires ETV5-mediated downstream activation of Brachyury in mice. Biol Reprod 85(6):1114–1123
Ishii K et al (2012) FGF2 mediates mouse spermatogonial stem cell self-renewal via upregulation of Etv5 and Bcl6b through MAP2K1 activation. Development 139(10):1734–1743
Zhang Y et al (2012) Endogenously produced FGF2 is essential for the survival and proliferation of cultured mouse spermatogonial stem cells. Cell Res 22(4):773–776
Boekhout M et al (2019) REC114 Partner ANKRD31 controls number, timing, and location of meiotic DNA breaks. Mol Cell 74(5):1053–1068 e8
Acknowledgments
This work was supported by National Institutes of Health (NIH) grant R01-HD103888 to N.W. X.Z. is supported by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences (NIGMS) of the NIH under grant number P20 GM103418 and KUMC Research Institute. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institute of General Medical Sciences, the National Institutes of Health, or KUMC Research Institute.
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Zhang, X., Wang, N. (2024). Induction of Meiotic Initiation in Long-Term Mouse Spermatogonial Stem Cells Under Retinoid Acid and Nutrient Restriction Conditions. In: Barchi, M., De Felici, M. (eds) Germ Cell Development. Methods in Molecular Biology, vol 2770. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3698-5_9
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DOI: https://doi.org/10.1007/978-1-0716-3698-5_9
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Publisher Name: Humana, New York, NY
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Online ISBN: 978-1-0716-3698-5
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