Department of Obstetrics and Gynecology, Feinberg School of Medicine, Northwestern University, Chicago, IL, USA.
Fertil Steril. 2013 Apr;99(5):1453-1459.e1. doi: 10.1016/j.fertnstert.2012.12.009. Epub 2013 Jan 8.
To determine whether the follicle environment modulates oocyte-specific gene transcript levels in cultured oocytes and polar bodies (PBs).
Animal study.
Large academic research center.
ANIMAL(S): CD1 mice.
INTERVENTION(S): In vitro growth of secondary mouse follicles in 0.25% or 1.5% alginate (ALG) in a three-dimensional culture system.
MAIN OUTCOME MEASURE(S): Relative transcript levels of Gdf9, Bmp15, Nlrp5, Tcl1, and Zp3 were measured by real-time quantitative reverse transcriptase-polymerase chain reaction in oocytes during in vitro follicle development and oocyte maturation and in their first PBs after removal from metaphase II (MII) eggs.
RESULT(S): All transcripts decreased earlier in oocytes cultured in 1.5% ALG compared with 0.25% ALG. Transcript levels were lower in MII eggs cultured in 1.5% ALG compared with in 0.25% ALG. All genes were expressed in PBs, and transcript levels were lower in PBs cultured in 1.5% ALG compared with in 0.25% ALG. Abundance of all transcripts was lower in PBs than in their sibling oocytes.
CONCLUSION(S): Local follicle environment modulates oocyte-specific gene expression in the oocyte and first PB. There is a significant difference in the transcript levels of oocyte-specific genes in PBs of 1.5% versus 0.25% ALG that correlates with ovarian environment-related decreases in oocyte competence.
确定卵泡环境是否调节培养的卵母细胞和极体中的卵母细胞特异性基因转录水平。
动物研究。
大型学术研究中心。
CD1 小鼠。
在三维培养系统中,0.25%或 1.5%藻酸盐(ALG)中二次小鼠卵泡的体外生长。
通过实时定量逆转录聚合酶链反应,测量卵母细胞在体外卵泡发育和卵母细胞成熟过程中以及从中期 II(MII)卵取出后的第一个极体中 Gdf9、Bmp15、Nlrp5、Tcl1 和 Zp3 的相对转录水平。
与 0.25%ALG 相比,1.5%ALG 培养的卵母细胞中所有转录物更早下降。与 0.25%ALG 相比,在 1.5%ALG 中培养的 MII 卵中的转录物水平较低。所有基因均在极体中表达,与在 0.25%ALG 中相比,在 1.5%ALG 中培养的极体中的转录物水平较低。与它们的姊妹卵母细胞相比,所有转录物在极体中的丰度均较低。
局部卵泡环境调节卵母细胞和第一极体中的卵母细胞特异性基因表达。在 1.5%与 0.25%ALG 相比,极体中卵母细胞特异性基因的转录水平存在显著差异,这与与卵巢环境相关的卵母细胞能力下降相关。