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Hum. Reprod. Advance Access published online on November 11, 2004
Human Reproduction, doi:10.1093/humrep/deh565
© 2004 by European Society of Human Reproduction and Embryology
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Received May 15, 2003
Revised July 22, 2004
Accepted September 20, 2004

Article

Molecular markers for the assessment of postnatal male germ cell development in the mouse

Sheba Jarvis 1, David J. Elliott 2, Delyth Morgan 1, Robert Winston 1, and Carol Readhead 3*

1 Institute of Reproductive and Developmental Biology, Imperial College Faculty of Medicine, Hammersmith Campus, Du Cane Road, London W12 ONN, UK
2 Institute of Human Genetics, The International Centre for Life, Central Parkway University of Newcastle-Upon-Tyne, NE1 3BZ, UK
3 The Biological Imaging Center, Beckman Institute 139-74, California Institute of Technology, Pasadena, CA 91125, USA

* To whom correspondence should be addressed.
Carol Readhead, E-mail: readhead{at}.caltech.edu


  Abstract

BACKGROUND: A proliferation marker, proliferating cell nuclear antigen (PCNA), a Sertoli cell specific transcription factor, GATA-1 and the male germ cell specific, RNA binding motif (RBM), were used to identify different cellular populations during postnatal development of the mouse testis. METHODS: Immunohistochemistry, RT-PCR and real-time quantitative RT-PCR (QRT-PCR) were used. RESULTS: PCNA was expressed in pre-Sertoli and germ cells on the day of birth. Both pre-meiotic germ cells and spermatocytes expressed RBM throughout postnatal development. RBM-positive cell counts and QRT-PCR of RBM showed that average level of RBM per cell is highest in juvenile males between 14 and 21 days. From 42 days onward, there was a dramatic decrease in RBM expression in individual pre-meiotic and meiotic germ cells. CONCLUSIONS: These markers were used to correlate cell proliferative capability, gene expression profile and anatomic location within the developing mouse testis. The majority of germ cells start active proliferation once they have migrated to the basement membrane or immediately before. RBM is more highly expressed during the first wave of spermatogenesis versus subsequent waves, suggesting that there may be a change in the activity of RBM.

Keywords: GATA-1; gonocytes; mouse spermatogenesis; proliferating cell nuclear antigen; RNA binding motif.
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