Ott T L, Zhou Y, Mirando M A, Stevens C, Harney J P, Ogle T F, Bazer F W
Department of Animal Science, University of Florida, Gainesville 32611.
J Mol Endocrinol. 1993 Apr;10(2):171-83. doi: 10.1677/jme.0.0100171.
This study characterized changes in levels of mRNA and protein for endometrial oestrogen receptors (ERs) and progesterone receptors (PRs) during luteolysis and maternal recognition of pregnancy. For cyclic and pregnant ewes, endometrium was collected on days 10, 12, 14, or 16 post-oestrus (4 ewes/day for each status) for the measurement of ER and PR mRNA and protein. The amount of receptor mRNA is expressed in relative units above background, measured from radiographs of dot-blot hybridization of total endometrial RNA with ER and PR cDNAs. At hysterectomy, jugular vein blood samples were collected and assayed for progesterone, total corpus luteum weight was recorded and, in vitro, endometrial oxytocin-stimulated inositol phosphate formation was estimated. In pregnant ewes, plasma progesterone increased gradually between days 10 and 16 (P < 0.01), corpus luteum weight was stable at approximately 0.8 g and oxytocin did not stimulate endometrial formation of inositol phosphates in vitro. In contrast, in cyclic ewes, plasma progesterone decreased from day 10 to day 16 (P < 0.01), corpus luteum weight decreased after day 14 to approximately 0.48 g (P = 0.05) and oxytocin stimulated an increase of approximately 1300% in the endometrial formation of inositol phosphates on day 16. cDNAs specifically hybridized with 1.6 and 3.1 kb transcripts for PR mRNA and a 6.5 kb transcript for ER mRNA. In cyclic ewes, the amount of PR mRNA increased from day 10 to maximum levels on days 14-16. The number of PRs decreased from day 10 (2.25 pmol/mg DNA) to day 12 (0.98 pmol/mg DNA) and then increased from day 14 to day 16 (2.8 pmol/mg DNA). In pregnant ewes, PR mRNA levels were greatest on days 10-12 and decreased by approximately 50% by day 16. In contrast, the number of PRs was relatively unchanged from day 10 to day 16 (1.53 to 1.03 pmol/mg DNA). In cyclic ewes, the amount of ER mRNA was lowest at day 10 and increased fivefold by day 16. The number of ERs remained relatively unchanged from day 10 to day 14 (6.07 pmol/mg DNA) and increased by day 16 (16.12 pmol/mg DNA). In pregnant ewes, ER mRNA decreased by approximately 80% from day 12 to day 16. Similarly, the number of ERs decreased from day 10 to day 16 (5.41 to 2.05 pmol/mg DNA). Correlations between ER mRNA and PR mRNA (r = 0.68), ERs and PRs (r = 0.50) and ER mRNA and ERs (r = 0.50) were high (P < 0.01).(ABSTRACT TRUNCATED AT 400 WORDS)
本研究对黄体溶解和母体识别妊娠期间子宫内膜雌激素受体（ERs）和孕激素受体（PRs）的mRNA和蛋白质水平变化进行了表征。对于发情周期和怀孕的母羊，在发情后第10、12、14或16天采集子宫内膜（每种状态每天4只母羊），用于测量ER和PR的mRNA和蛋白质。受体mRNA的量以高于背景的相对单位表示，通过将总子宫内膜RNA与ER和PR cDNA进行斑点印迹杂交的放射自显影片测量得出。在子宫切除时，采集颈静脉血样并测定孕酮，记录黄体总重量，并在体外估计子宫内膜催产素刺激的肌醇磷酸形成。在怀孕母羊中，血浆孕酮在第10至16天逐渐增加（P<0.01），黄体重量稳定在约0.8 g，催产素在体外不刺激子宫内膜形成肌醇磷酸。相比之下，在发情周期的母羊中，血浆孕酮从第10天降至第16天（P<0.01），黄体重量在第14天后降至约0.48 g（P = 0.05），催产素在第16天刺激子宫内膜肌醇磷酸形成增加约1300%。cDNA分别与1.6和3.1 kb的PR mRNA转录本以及6.5 kb的ER mRNA转录本特异性杂交。在发情周期的母羊中，PR mRNA的量从第10天增加到第14 - 16天的最高水平。PR的数量从第10天（2.25 pmol/mg DNA）降至第12天（0.98 pmol/mg DNA），然后从第14天增加到第16天（2.8 pmol/mg DNA）。在怀孕母羊中，PR mRNA水平在第10 - 12天最高，到第16天下降约50%。相比之下，PR的数量从第10天到第16天相对不变（1.53至1.03 pmol/mg DNA）。在发情周期的母羊中，ER mRNA的量在第10天最低，到第16天增加了五倍。ER的数量从第10天到第14天相对不变（6.07 pmol/mg DNA），到第16天增加（16.12 pmol/mg DNA）。在怀孕母羊中，ER mRNA从第12天到第16天下降约80%。同样，ER的数量从第10天到第16天下降（5.41至2.05 pmol/mg DNA）。ER mRNA与PR mRNA（r = 0.68）、ER与PR（r = 0.50）以及ER mRNA与ER（r = 0.50）之间的相关性很高（P<0.01）。（摘要截断于400字）
