Molecular markers for the assessment of postnatal male germ cell development in the mouse.
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.
Jarvis
Sheba
S
Institute of Reproductive and Developmental Biology, Imperial College Faculty of Medicine, Hammersmith Campus, Du Cane Road, London W12 ONN, UK.
Elliott
David J
DJ
Morgan
Delyth
D
Winston
Robert
R
Readhead
Carol
C
D000818:Animals; D000831:Animals, Newborn; D001483:Base Sequence; D015415:Biomarkers / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics; D004268:DNA-Binding Proteins / Q000378:metabolism; D050981:Erythroid-Specific DNA-Binding Factors; D050982:GATA1 Transcription Factor; D018507:Gene Expression Regulation, Developmental; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009687:Nuclear Proteins; D018809:Proliferating Cell Nuclear Antigen / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012708:Sertoli Cells / Q000378:metabolism; D013091:Spermatogenesis / Q000235:genetics / Q000502:physiology; D013094:Spermatozoa / Q000166:cytology / Q000378:metabolism*; D013737:Testis / Q000166:cytology / Q000254:growth & development* / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism
D015415:Biomarkers; D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D050981:Erythroid-Specific DNA-Binding Factors; D050982:GATA1 Transcription Factor; C494689:Gata1 protein, mouse; D009687:Nuclear Proteins; D018809:Proliferating Cell Nuclear Antigen; D016601:RNA-Binding Proteins; C501500:Rbmy protein, mouse; D014157:Transcription Factors