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催乳素相关激素在早期牛孕体中的表达及其对子宫内膜旁分泌作用的可能性。

Expression of prolactin-related hormones in the early bovine conceptus, and potential for paracrine effect on the endometrium.

作者信息

Kessler M A, Duello T M, Schuler L A

机构信息

Department of Comparative Biosciences, University of Wisconsin, Madison 53706.

出版信息

Endocrinology. 1991 Oct;129(4):1885-95. doi: 10.1210/endo-129-4-1885.

Abstract

Hormones related to the pituitary hormones PRL and GH are produced by the utero-placental unit of many species. In the cow, these include bovine placental lactogen (bPL) and a distantly related subfamily including the protein encoded by bovine PRL-related complementary DNA I (bPRCI). In the present studies, we defined the onset of expression of these genes in order to begin to study the regulation of their expression and function before implantation. Messenger RNA levels of both bPL and bPRCI were assessed by dot blot analysis of total RNAs prepared from conceptuses on days 15-25. Specific transcripts were not detectable at day 15 but were readily apparent beginning at day 17. To define the portions of the placenta responsible for expressing these genes, RNA was prepared from chorion (separated into cotyledons and intercotyledonary membrane), allantois, and amnion from day 58 gestation and analyzed by Northern hybridization. Transcripts for both these hormones were confined to the chorion, with intercotyledonary RNA containing at least as much as that prepared from cotyledons. To localize the product of these genes, extraembryonic membranes were immunostained with bPL and bPRCI antisera. Mononucleated trophoblastic cells stained for bPL and bPRCI at day 18; similar staining was apparent at days 23 and 24, as well as in some but not all binucleated cells. Microsomes prepared from endometrium exhibited specific binding of bPL throughout pregnancy as well as during the luteal phase of the cycle. At midgestation, these high affinity, low capacity receptors (dissociation constant of 1.27 x 10(-10) M, maximum binding capacity of 44.2 fmol/mg protein) demonstrated a selective affinity for bPL in preference to bGH and bPRL. Using this radioreceptor assay, we demonstrated the presence of bPL at nanomolar concentrations in washings from the uterine lumen, suggesting that bPL may exert paracrine effects on the endometrium.

摘要

许多物种的子宫 - 胎盘单位会产生与垂体激素催乳素(PRL)和生长激素(GH)相关的激素。在牛中,这些激素包括牛胎盘催乳素(bPL)以及一个与之亲缘关系较远的亚家族,其中包括由牛PRL相关互补DNA I(bPRCI)编码的蛋白质。在本研究中,我们确定了这些基因的表达起始时间,以便在着床前开始研究它们的表达调控和功能。通过对第15 - 25天的孕体所制备的总RNA进行斑点印迹分析,评估了bPL和bPRCI的信使RNA水平。在第15天未检测到特异性转录本,但从第17天开始就很明显了。为了确定负责表达这些基因的胎盘部分,从妊娠第58天的绒毛膜(分为子叶和子叶间膜)、尿囊和羊膜制备RNA,并通过Northern杂交进行分析。这两种激素的转录本都局限于绒毛膜,子叶间RNA的含量至少与从子叶制备的RNA一样多。为了定位这些基因的产物,用bPL和bPRCI抗血清对胚外膜进行免疫染色。在第18天,单核滋养层细胞对bPL和bPRCI染色;在第23天和第24天以及一些但不是所有的双核细胞中也有类似的染色。从子宫内膜制备的微粒体在整个妊娠期间以及周期的黄体期都表现出bPL的特异性结合。在妊娠中期,这些高亲和力、低容量的受体(解离常数为1.27×10⁻¹⁰ M,最大结合容量为44.2 fmol/mg蛋白质)对bPL表现出比对bGH和bPRL的选择性亲和力。使用这种放射受体测定法,我们证明了子宫腔冲洗液中存在纳摩尔浓度的bPL,这表明bPL可能对子宫内膜发挥旁分泌作用。

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