Abstract
Germ cells make two major decisions when they move from an indeterminate state to their final stage of gamete production. One decision is sexual commitment for sperm or egg production, and the other is to maintain mitotic division or entry into meiosis. It is unclear whether the two decisions are made as a single event or separate events, because there has been no evidence for the presence of germ cell sex prior to meiosis. Here we report direct evidence in the fish rainbow trout that gonia have distinct sexuality. We show that dazl expression occurs in both male and female gonia but exhibits differential intracellular distribution. More strikingly, we show that boule is highly expressed in male gonia but absent in female gonia. Therefore, mitotic gonia possess sex, sperm/egg decision and mitosis/meiosis decision are two independent events, and sperm/egg decision precedes mitosis/meiosis decision in rainbow trout, making this organism a unique vertebrate model for mechanistic understanding of germ cell sex differentiation and relationship between the two decisions.
Keywords: boule, dazl, germ cell sex, gonia, meiosis, trout
Footnotes
These authors contributed equally to this work.
References
- Baron D., Houlgatte R., Fostier A., Guiguen Y. Largescale temporal gene expression profiling during gonadal differentiation and early gametogenesis in rainbow trout. Biol Reprod. 2005;73:959–966. doi: 10.1095/biolreprod.105.041830. [DOI] [PubMed] [Google Scholar]
- Campbell K.H., Fisher P., Chen W.C., Choi I., Kelly R.D., Lee J.H., Xhu J. Somatic cell nuclear transfer: Past, present and future perspectives. Theriogenology. 2007;68:S214–S231. doi: 10.1016/j.theriogenology.2007.05.059. [DOI] [PubMed] [Google Scholar]
- Herpin A., Rohr S., Riedel D., Kluever N., Raz E., Schartl M. Specification of primordial germ cells in medaka (Oryzias latipes) BMC Dev Biol. 2007;7:3. doi: 10.1186/1471-213X-7-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hong Y., Liu T., Zhao H., Xu H., Wang W., Liu R., Chen T., Deng J., Gui J. Establishment of a normal medakafish spermatogonial cell line capable of sperm production in vitro. Proc Natl Acad Sci U S A. 2004;101:8011–8016. doi: 10.1073/pnas.0308668101. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hong Y., Winkler C., Schartl M. Pluripotency and differentiation of embryonic stem cell lines from the medakafish (Oryzias latipes) Mech Dev. 1996;60:33–44. doi: 10.1016/S0925-4773(96)00596-5. [DOI] [PubMed] [Google Scholar]
- Hong Y., Winkler C., Schartl M. Production of medakafish chimeras from a stable embryonic stem cell line. Proc Natl Acad Sci U S A. 1998;95:3679–3684. doi: 10.1073/pnas.95.7.3679. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kimble J., Page D.C. The mysteries of sexual identity. The germ cell’s perspective. Science. 2007;316:400–401. doi: 10.1126/science.1142109. [DOI] [PubMed] [Google Scholar]
- Kocer A., Reichmann J., Best D., Adams I.R. Germ cell sex determination in mammals. Mol Hum Reprod. 2009;15:205–213. doi: 10.1093/molehr/gap008. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kondo M., Nanda I., Hornung U., Asakawa S., Shimizu N., Mitani H., Schmid M., Shima A., Schartl M. Absence of the candidate male sex-determining gene dmrt1b(Y) of medaka from other fish species. Curr Biol. 2003;13:416–420. doi: 10.1016/S0960-9822(03)00125-8. [DOI] [PubMed] [Google Scholar]
- Kurokawa H., Saito D., Nakamura S., Katoh-Fukui Y., Ohta K., Baba T., Morohashi K., Tanaka M. Germ cells are essential for sexual dimorphism in the medaka gonad. Proc Natl Acad Sci U S A. 2007;104:16958–16963. doi: 10.1073/pnas.0609932104. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Li M., Hong N., Xu H., Yi M., Li C., Gui J., Hong Y. Medaka vasa is required for migration but not survival of primordial germ cells. Mech Dev. 2009;126:366–381. doi: 10.1016/j.mod.2009.02.004. [DOI] [PubMed] [Google Scholar]
- Li, M., Shen, Q., Xu, H., Wong, F.M., Cui, J., Li, Z., Hong, N., Wang, L., Zhao, H., Ma, B., et al. (2011). Differential conservation and divergence of fertility genes boule and dazl in the rainbow trout. PloS ONE. (In press) [DOI] [PMC free article] [PubMed]
- Liu L., Hong N., Xu H., Li M., Yan Y., Purwanti Y., Yi M., Li Z., Wang L., Hong Y. Medaka dead end encodes a cytoplasmic protein and identifies embryonic and adult germ cells. Gene Expr Patterns. 2009;9:541–548. doi: 10.1016/j.gep.2009.06.008. [DOI] [PubMed] [Google Scholar]
- Matsuda M., Nagahama Y., Shinomiya A., Sato T., Matsuda C., Kobayashi T., Morrey C.E., Shibata N., Asakawa S., Shimizu N., et al. DMY is a Y-specific DM-domain gene required for male development in the medaka fish. Nature. 2002;417:559–563. doi: 10.1038/nature751. [DOI] [PubMed] [Google Scholar]
- Nakamoto M., Suzuki A., Matsuda M., Nagahama Y., Shibata N. Testicular type Sox9 is not involved in sex determination but might be in the development of testicular structures in the medaka, Oryzias latipes. Biochem Biophys Res Commun. 2005;333:729–736. doi: 10.1016/j.bbrc.2005.05.158. [DOI] [PubMed] [Google Scholar]
- Nanda I., Kondo M., Hornung U., Asakawa S., Winkler C., Shimizu A., Shan Z., Haaf T., Shimizu N., Shima A., et al. A duplicated copy of DMRT1 in the sex-determining region of the Y chromosome of the medaka, Oryzias latipes. Proc Natl Acad Sci U S A. 2002;99:11778–11783. doi: 10.1073/pnas.182314699. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Okutsu T., Shikina S., Kanno M., Takeuchi Y., Yoshizaki G. Production of trout offspring from triploid salmon parents. Science. 2007;317:1517. doi: 10.1126/science.1145626. [DOI] [PubMed] [Google Scholar]
- Okutsu T., Suzuki K., Takeuchi Y., Takeuchi T., Yoshizaki G. Testicular germ cells can colonize sexually undifferentiated embryonic gonad and produce functional eggs in fish. Proc Natl Acad Sci U S A. 2006;103:2725–2729. doi: 10.1073/pnas.0509218103. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Uhlenhaut N.H., Jakob S., Anlag K., Eisenberger T., Sekido R., Kress J., Treier A.C., Klugmann C., Klasen C., Holter N.I., et al. Somatic sex reprogramming of adult ovaries to testes by FOXL2 ablation. Cell. 2009;139:1130–1142. doi: 10.1016/j.cell.2009.11.021. [DOI] [PubMed] [Google Scholar]
- Vizziano D., Randuineau G., Baron D., Cauty C., Guiguen Y. Characterization of early molecular sex differentiation in rainbow trout, Oncorhynchus mykiss. Dev Dyn. 2007;236:2198–2206. doi: 10.1002/dvdy.21212. [DOI] [PubMed] [Google Scholar]
- Volff J.N., Nanda I., Schmid M., Schartl M. Governing sex determination in fish: regulatory putsches and ephemeral dictators. Sex Dev. 2007;1:85–99. doi: 10.1159/000100030. [DOI] [PubMed] [Google Scholar]
- Wylie C. Germ cells. Cell. 1999;96:165–174. doi: 10.1016/S0092-8674(00)80557-7. [DOI] [PubMed] [Google Scholar]
- Xu H., Gui J., Hong Y. Differential expression of vasa RNA and protein during spermatogenesis and oogenesis in the gibel carp (Carassius auratus gibelio), a bisexually and gynogenetically reproducing vertebrate. Dev Dyn. 2005;233:872–882. doi: 10.1002/dvdy.20410. [DOI] [PubMed] [Google Scholar]
- Xu H., Li M., Gui J., Hong Y. Cloning and expression of medaka dazl during embryogenesis and gametogenesis. Gene Expr Patterns. 2007;7:332–338. doi: 10.1016/j.modgep.2006.08.001. [DOI] [PubMed] [Google Scholar]
- Xu H., Li M., Gui J., Hong Y. Fish germ cells. Sci China Life Sci. 2010;53:435–446. doi: 10.1007/s11427-010-0058-8. [DOI] [PubMed] [Google Scholar]
- Xu H., Li Z., Li M., Wang L., Hong Y. Boule is present in fish and bisexually expressed in adult and embryonic germ cells of medaka. PLoS One. 2009;4:e6097. doi: 10.1371/journal.pone.0006097. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yi M., Hong N., Hong Y. Generation of medaka fish haploid embryonic stem cells. Science. 2009;326:430–433. doi: 10.1126/science.1175151. [DOI] [PubMed] [Google Scholar]
- Yoshizaki G., Ichikawa M., Hayashi M., Iwasaki Y., Miwa M., Shikina S., Okutsu T. Sexual plasticity of ovarian germ cells in rainbow trout. Development. 2010;137:1227–1230. doi: 10.1242/dev.044982. [DOI] [PubMed] [Google Scholar]
