[Meiosis related gene expression in rat spermatogenesis].
OBJECTIVE
To find the expression of specific genes related to the meiosis of germ cells during spermatogenesis in the rat testis.


METHODS
Segments of seminiferous tubules were obtained from the adult male SD rats, at stages XIII - I of meiosis, and the interstitial cells of the same testis were isolated under the stereomicroscope. The total RNAs of stages XIII - I segments and the testicular interstitial cells were extracted respectively, and mRNA differential display RT-PCR (DDRT-PCR) was conducted. The obtained cDNA fractions were purified and recovered, the reverse dot blot hybridization, sub-clones and screens of blue/white dots performed, and the results of sub-clones were identified by restriction endonuclease EcoR I digestion.


RESULTS
Sixteen differential cDNA fractions were obtained through primary DDRT-PCR, 7 from stages XIII - I segments and 9 from the testicular interstitial cells. Another 11 were selected for further screening by reverse dot blot hybridization, their size ranging from 200 to 500 bp, of which 6 were from the stages XIII - I segments of seminiferous tubules and the other 5 from the rat testicular interstitial cells. All of the 11 cDNA fractions were sub-cloned and screened by blue/white dots.


CONCLUSIONS
Specifically expressed differential cDNA fractions can be obtained and primarily identified from testicular interstitial cells and the seminiferous tubules, which, as the sequence tags of the testicular meiotic expression, deserve further investigation.
Liu
De-Yu
DY
National Research Institute for Population and Family Planning, Beijing 100081, China. liudeyu222@yahoo.com.cn
Wu
Yan-Wan
YW
D000818:Animals; D020224:Expressed Sequence Tags; D018507:Gene Expression Regulation, Developmental; D007985:Leydig Cells / Q000166:cytology; D008297:Male; D008540:Meiosis / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012671:Seminiferous Tubules / Q000166:cytology; D013091:Spermatogenesis / Q000235:genetics*