Jarvis Sheba, Elliott David J, Morgan Delyth, Winston Robert, Readhead Carol
Institute of Reproductive and Developmental Biology, Imperial College Faculty of Medicine, Hammersmith Campus, Du Cane Road, London W12 ONN, UK.
Hum Reprod. 2005 Jan;20(1):108-16. doi: 10.1093/humrep/deh565. Epub 2004 Nov 11.
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.
Immunohistochemistry, RT-PCR and real-time quantitative RT-PCR (QRT-PCR) were used.
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.
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.
增殖标志物增殖细胞核抗原（PCNA）、支持细胞特异性转录因子GATA-1以及雄性生殖细胞特异性RNA结合基序（RBM），被用于鉴定小鼠睾丸出生后发育过程中的不同细胞群体。
采用免疫组织化学、逆转录聚合酶链反应（RT-PCR）和实时定量逆转录聚合酶链反应（QRT-PCR）。
出生当天，PCNA在支持细胞前体细胞和生殖细胞中表达。减数分裂前生殖细胞和精母细胞在整个出生后发育过程中均表达RBM。RBM阳性细胞计数和RBM的QRT-PCR显示，14至21天的幼年雄性小鼠中，每个细胞的RBM平均水平最高。从42天起，单个减数分裂前和减数分裂生殖细胞中的RBM表达急剧下降。
这些标志物被用于关联发育中小鼠睾丸内的细胞增殖能力、基因表达谱和解剖位置。大多数生殖细胞一旦迁移到基底膜或在此之前就开始活跃增殖。与后续波次相比，RBM在第一波精子发生过程中表达更高，这表明RBM的活性可能发生了变化。
