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Human DAZL, DAZ and BOULE genes modulate primordial germ-cell and haploid gamete formation

Nature volume 462, pages 222–225 (2009)Cite this article

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Abstract

The leading cause of infertility in men and women is quantitative and qualitative defects in human germ-cell (oocyte and sperm) development. Yet, it has not been possible to examine the unique developmental genetics of human germ-cell formation and differentiation owing to inaccessibility of germ cells during fetal development. Although several studies have shown that germ cells can be differentiated from mouse and human embryonic stem cells, human germ cells differentiated in these studies generally did not develop beyond the earliest stages1,2,3,4,5,6,7,8. Here we used a germ-cell reporter to quantify and isolate primordial germ cells derived from both male and female human embryonic stem cells. By silencing and overexpressing genes that encode germ-cell-specific cytoplasmic RNA-binding proteins (not transcription factors), we modulated human germ-cell formation and developmental progression. We observed that human DAZL (deleted in azoospermia-like) functions in primordial germ-cell formation, whereas closely related genes DAZ and BOULE (also called BOLL) promote later stages of meiosis and development of haploid gametes. These results are significant to the generation of gametes for future basic science and potential clinical applications.

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Figure 1: Enrichment of human germ cells by BMPs and VASA–GFP reporter.
Figure 2: Germ-cell properties of VASA–GFP + cells.
Figure 3: Silencing of DAZ family members and germ-cell numbers.
Figure 4: Overexpression of DAZL, DAZ and BOULE induces meiotic progression and haploid formation.

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  • 12 November 2009

    A definition for the scale bars in the legend for Fig. 2b was added on 12 November 2009.

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Acknowledgements

We thank D. Suter for the p2k7 vectors, B. Behr for procurement of clinical samples, C. Nicholas for assistance with SCP3 staining, and S. Jiang and P. Lovelace for FACS expertise. K.K. was supported by a California TRDRP postdoctoral fellowship; V.T.A. was supported by an NIH fellowship. This work was supported by the National Institutes of Health (RO1HD047721, U54HD055764 as part of the Specialized Cooperative Centers Program in Reproduction and Infertility Research), the California TRDRP (14RT0159) and the California Institute for Regenerative Medicine (RC1-00137) to R.A.R.P.

Author Contributions K.K. carried out the majority of experiments with assistance with imprint analysis and gene expression analysis by V.T.A., silencing of BOULE and DAZ by M.F., and 5mC and FISH staining by H.N.N.; K.K. and R.A.R.P. designed experiments and wrote the manuscript.

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Authors and Affiliations

  1. Department of Obstetrics and Gynecology, Center for Human Embryonic Stem Cell Research and Education, Institute for Stem Cell Biology & Regenerative Medicine, Stanford University School of Medicine, Stanford University, Palo Alto, California 94305, USA,

    Kehkooi Kee, Vanessa T. Angeles, Martha Flores, Ha Nam Nguyen & Renee A. Reijo Pera

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  1. Kehkooi Kee
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  2. Vanessa T. Angeles
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Correspondence to Renee A. Reijo Pera.

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Kee, K., Angeles, V., Flores, M. et al. Human DAZL, DAZ and BOULE genes modulate primordial germ-cell and haploid gamete formation . Nature 462, 222–225 (2009). https://doi.org/10.1038/nature08562

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