Wu W X, Verbalis J G, Hoffman G E, Derks J B, Nathanielsz P W
Department of Physiology, Cornell University College of Veterinary Medicine, Ithaca, New York 14853-6401, USA.
Endocrinology. 1996 Feb;137(2):722-8. doi: 10.1210/endo.137.2.8593823.
At the end of pregnancy, the myometrium becomes extremely sensitive to oxytocin (OT) as result of a dramatic increase in the number of OT receptors (OTR), indicating an important role for OTR in the process of labor. There are no studies in sheep in which the physical properties and histological distribution of OTR are evaluated in relation to parturition. Also, no studies have been performed in any species to simultaneously examine the distribution of OTR at the messenger RNA (mRNA) as well as the protein levels in the same tissues and correlate those changes with the patterns of myometrial activity that occur at labor. In the present studies, we have used a polyclonal anti-OTR antibody and Western blot analysis to determine the apparent molecular mass of ovine OTR in late pregnant sheep myometrium and endometrium. We also examined the distribution of OTR mRNA and protein expression in the intact myometrium and endometrium and in individual cultured cells using in situ hybridization and immunocytochemistry. The expression of OTR and its mRNA has been correlated with the patterns of activity observed in the pregnant sheep myometrium. Western blot analysis of myometrial and endometrial extracts revealed a major form of OTR with an approximate molecular mass of 66 kDa. Both immunocytochemistry and in situ hybridization localized OTR and its mRNA in myometrial cells and glandular cells of the endometrium. Increased OTR and its mRNA expression in the myometrium and endometrium were correlated with the occurrence of myometrial contractions. OTR was also demonstrated by immunocytochemistry in the smooth muscle of myometrial blood vessels. Localization of OTR and its mRNA in pregnant sheep myometrial cells is consistent with the hypothesis that OTR plays an important role in regulating myometrial contractility. Positive staining of OTR in endometrial glandular cells supports the view that OT is involved in PG production by the endometrium in late pregnancy. Increased expression of OTR and its mRNA in the myometrium during labor further indicates that changes in tissue OTR play a significant role in the mechanism of parturition. Increased expression of OTR and its mRNA in endometrium may relate to the role of OT in regulating PG production by the endometrium during labor.
在妊娠末期,由于催产素(OT)受体(OTR)数量急剧增加,子宫肌层对OT变得极其敏感,这表明OTR在分娩过程中起重要作用。目前尚无关于绵羊OTR的物理特性和组织学分布与分娩关系的研究。此外,在任何物种中,均未进行过同时检测同一组织中OTR在信使核糖核酸(mRNA)水平和蛋白质水平的分布,并将这些变化与分娩时子宫肌层活动模式相关联的研究。在本研究中,我们使用多克隆抗OTR抗体和蛋白质印迹分析来确定妊娠晚期绵羊子宫肌层和子宫内膜中绵羊OTR的表观分子量。我们还使用原位杂交和免疫细胞化学方法,检测了完整子宫肌层和子宫内膜以及单个培养细胞中OTR mRNA和蛋白质表达的分布情况。OTR及其mRNA的表达已与妊娠绵羊子宫肌层中观察到的活动模式相关联。子宫肌层和子宫内膜提取物的蛋白质印迹分析显示,主要的OTR形式的分子量约为66 kDa。免疫细胞化学和原位杂交均将OTR及其mRNA定位在子宫肌层细胞和子宫内膜的腺细胞中。子宫肌层和子宫内膜中OTR及其mRNA表达的增加与子宫肌层收缩的发生相关。免疫细胞化学还在子宫肌层血管的平滑肌中证实了OTR的存在。OTR及其mRNA在妊娠绵羊子宫肌层细胞中的定位与OTR在调节子宫肌层收缩性中起重要作用的假说一致。OTR在子宫内膜腺细胞中的阳性染色支持了OT参与妊娠晚期子宫内膜前列腺素(PG)产生的观点。分娩期间子宫肌层中OTR及其mRNA表达的增加进一步表明,组织OTR的变化在分娩机制中起重要作用。子宫内膜中OTR及其mRNA表达的增加可能与OT在分娩期间调节子宫内膜PG产生的作用有关。