Menkhorst E, Selwood L, Cui S
Department of Zoology, The University of Melbourne, Royal Parade, Parkville, VIC 3010, Australia.
Mol Reprod Dev. 2009 Sep;76(9):863-72. doi: 10.1002/mrd.21053.
The marsupial conceptus is surrounded by a uterine-secreted shell coat for 60-80% of gestation. Coat protein 4 (CP4) is the only marsupial shell coat protein characterized and it has only been identified in one species, the Common Brushtail Possum. In this possum, uterine transcription and secretion of cp4 during the oestrous cycle is biphasic and associated with the stage of conceptus development. Here we cloned cp4 (sm-cp4) from a distantly related species, the Stripe-faced Dunnart (Sminthopsis macroura). Transcription of sm-cp4 and secretion of smCP4 were identified by semi-quantitative RT-PCR and immunohistochemistry, respectively. The effect of reproductive hormones on sm-cp4 transcription was investigated in vitro by treatment of uterine explant cultures with oestrogen and/or progesterone. In vivo, uterine expression of smCP4 was biphasic and associated with conceptus development. Uterine smCP4 expression (transcription and secretion) began during the pre-ovulatory period and continued post-ovulation during cleavage stages. Transcription of sm-cp4 continued during the unilaminar blastocyst stage, but smCP4 secretion was reduced during this stage. During the bilaminar blastocyst stage, both transcription of sm-cp4 and secretion of smCP4 were low before they both resumed during the trilaminar blastocyst stage, and continued during the embryo and fetal stages. In vitro uterine transcription of sm-cp4 increased after incubation with progesterone.
有袋类动物的孕体在妊娠期的60%-80%时间里被子宫分泌的壳膜所包裹。壳膜蛋白4(CP4)是唯一已被鉴定的有袋类动物壳膜蛋白,且仅在一种动物——帚尾袋貂中被识别出来。在这种袋貂中,发情周期中cp4的子宫转录和分泌是双相的,且与孕体发育阶段相关。在此,我们从亲缘关系较远的细纹袋鼬(Sminthopsis macroura)中克隆了cp4(sm-cp4)。分别通过半定量RT-PCR和免疫组织化学鉴定了sm-cp4的转录和smCP4的分泌。通过用雌激素和/或孕酮处理子宫外植体培养物,在体外研究了生殖激素对sm-cp4转录的影响。在体内,子宫smCP4的表达是双相的,且与孕体发育相关。子宫smCP4的表达(转录和分泌)在排卵前期开始,并在排卵后卵裂阶段持续。sm-cp4的转录在单层胚泡阶段继续,但在此阶段smCP4的分泌减少。在双层胚泡阶段,sm-cp4的转录和smCP4的分泌在三层胚泡阶段恢复之前均较低,之后在胚胎和胎儿阶段继续。与孕酮孵育后,体外子宫sm-cp4的转录增加。