【摘要】 利用免疫组织化学及RT PCR的方法 ,研究了细胞周期蛋白D3(CyclinD3)在兔早期妊娠子宫和着床前胚胎中的表达情况 ,以揭示CyclinD3在兔胚胎着床过程中的可能作用。结果显示 :( 1)在兔妊娠第 3～ 8天的子宫近肌层的腺上皮中有CyclinD3免疫染色 ,并且其表达强度在第 7天以后呈现下降的趋势 ,着床的胚胎中未见CyclinD3免疫染色 ;( 2 )在第 2～ 5天的假孕兔子宫的腺上皮中有较强的CyclinD3免疫染色 ;( 3)在发情周期的兔子宫中未见其有表达 ;( 4 )在切除卵巢的兔中 ,注射雌激素后子宫中未见CyclinD3免疫染色 ,注射孕酮后在子宫腺上皮中有CyclinD3免疫染色 ,在孕酮和雌激素共同处理后的子宫腺上皮中有较强的CyclinD3免疫染色 ;( 5 )利用RT PCR的方法在早期胚胎中均能检测到CyclinD3,但从胚泡期开始表达上升 ,到扩展胚泡时其表达最强。上述结果表明 ,CyclinD3的表达可能对兔胚泡的着床具有一定的调节作用。更多还原

【Abstract】 Cyclin D3, an important factor in the regulation of the G1/S transition of the cell cycle in various cell types cultured in vitro , can maintain the balance of cell growth and development. Although cyclin D3 is expressed in mouse decidualized stromal cells after implantation, cyclin D3 expression in the uterus of other species has not been reported. The aim of this study was to investigate the expression and regulation of cyclin D3 in rabbit uterus and preimplantation embryos during early pregnancy by immunohistochemistry and reverse transcription PCR (RT PCR). Cyclin D3 expression was also examined in pseudopregnant and steroid treated rabbit uterus. There was no detectable cyclin D3 immunostaining in the uteri on day 1 and 2 of pregnancy. In the uteri on day 3 to 6 of pregnancy, a strong immunostaining of cyclin D3 was seen in the glandular epithelium, while no immunostaining was observed in the luminal epithelium. Cyclin D3 immunostaining was detected in the glandular epithelium at implantation site on day 7 of pregnancy, being stronger on the anti mesometrial side than that on the mesometrial side. There was detectable cyclin D3 immunostaining in the glandular epithelium of the inter implantation area on day 7 of pregnancy. On day 8 of pregnancy, a week immunostaining of cyclin D3 was seen only in the glandular epithelium on the anti mesometrial side. Cyclin D3 mRNA expression was detected in the uterine endometrium from days 1 to 8 of pregnancy by RT PCR analysis. A high level of cyclin D3 mRNA was only seen in the endometrium from days 4 to 8 of pregnancy. In pseudopregnant uteri, no cyclin D3 immunostaining was seen on days 1 and 6, while a strong immunostaining of cyclin D3 was present in the glandular epithelium on days 2-5. In rabbit uteri at estrus and diestrus, cyclin D3 immunostaining was undetectable. There was no cyclin D3 immunostaining in the ovariectomized uterus. Cyclin D3 was induced in the glandular epithelium after progesterone treatment, but no immunostaining was detected after estrogen treatment. A combination of estrogen and progesterone also stimulated cyclin D3 immunostaining in the glandular epithelium of ovariectomized rabbit. By RT PCR analysis, cyclin D3 mRNA was detected in oocytes and all the stages of preimplantation embryos, and was highly expressed in the expanded and hatched blastocysts. These results suggest that the expression of cyclin D3 may play an important role during rabbit blastocyst implantation.更多还原