Reproductive Biology Research Unit, Division of Animal Sciences, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305-8602, Japan.
Reprod Biol Endocrinol. 2011 Mar 15;9:33. doi: 10.1186/1477-7827-9-33.
It has been reported that calf oocytes are less developmentally competent than oocytes obtained from adult cows. Bone morphogenetic protein 15 (BMP15) and growth and differentiation factor 9 (GDF9) play critical roles in folliculogenesis, follicular development and ovulation in mammalian ovaries. In the present study, we attempted to compare the expression patterns of BMP15 and GDF9 in the cells of calf and cow ovaries to determine a relationship between the level of these genes and the low developmental competence of calf oocytes.
Bovine tissues were collected from 9-11 months-old calves and from 4-6 years-old cows. We characterized the gene expression of BMP15 and GDF9 in calf and adult bovine oocytes and cumulus cells using quantitative real-time reverse transcriptase polymerase chain reaction (QPCR) and in situ hybridization. Immunohistochemical analysis was also performed.
The expression of BMP15 and GDF9 in cumulus cells of adult ovaries was significantly higher than that in calf ovaries, as revealed by QPCR. GDF9 expression in the oocytes of calf ovaries was significantly higher than in those of the adult ovaries. In contrast, BMP15 expression in the oocytes of calf and adult ovaries was not significantly different. The localization of gene expression and protein were ascertained by histochemistry.
Our result showed for the first time BMP15 and GDF9 expression in bovine cumulus cells. BMP15 and GDF9 mRNA expression in oocytes and cumulus cells was different in calves and cows.
据报道,小牛卵母细胞的发育能力不如成年牛获得的卵母细胞。骨形态发生蛋白 15(BMP15)和生长分化因子 9(GDF9)在哺乳动物卵巢的卵泡发生、卵泡发育和排卵中发挥着关键作用。在本研究中,我们试图比较小牛和奶牛卵巢中 BMP15 和 GDF9 细胞的表达模式,以确定这些基因的水平与小牛卵母细胞发育能力低之间的关系。
从小牛(9-11 月龄)和成年奶牛(4-6 岁)中收集牛组织。我们使用定量实时逆转录聚合酶链反应(QPCR)和原位杂交技术,对小牛和成年牛卵母细胞和卵丘细胞中 BMP15 和 GDF9 的基因表达进行了表征。还进行了免疫组织化学分析。
QPCR 结果显示,成年卵巢中卵丘细胞的 BMP15 和 GDF9 表达明显高于小牛卵巢。小牛卵巢卵母细胞的 GDF9 表达明显高于成年卵巢。相比之下,小牛和成年卵巢卵母细胞的 BMP15 表达没有显著差异。通过组织化学确定了基因表达和蛋白质的定位。
本研究首次显示了 BMP15 和 GDF9 在牛卵丘细胞中的表达。小牛和奶牛的卵母细胞和卵丘细胞中的 BMP15 和 GDF9 mRNA 表达不同。