Olofsson J I, Leung C H, Bjurulf E, Ohno T, Selstam G, Peng C, Leung P C
Department of Physiology, University of Umeå, Sweden.
Mol Cell Endocrinol. 1996 Oct 14;123(1):45-52. doi: 10.1016/0303-7207(96)03894-4.
The recent cloning of several cDNAs encoding prostaglandin (PG) receptors has paved the way for a more detailed investigation of the postulated regulatory role of prostaglandins in corpus luteum function. We have utilized the reverse transcription-polymerase chain reaction (RT-PCR) to isolate a mRNA encoding the ovarian PGF(2alpha) (FP) receptor, using oligonucleotides based on the recently cloned mouse cDNA as primers. The 5'-untranslated region of the rat ovarian mRNA was isolated following 5'-RACE (rapid amplification of cDNA ends). The isolated 1526 base-pair sequence, which spans the entire open reading frame, was found 100% identical in the protein coding region to a similar sequence isolated from a rat astrocyte cDNA library, but different in the first 32 nucleotides of the 5'-untranslated region, possibly due to tissue-specific splicing heterogeneity. Using ribonuclease protection assay, a quantitative analysis of FP receptor mRNA levels was performed in corpora lutea excised from adult pseudopregnant rats (Day 8) at different timepoints (0.5-48 h) following the in vivo s.c. regimen of a luteolytic dose of the FP receptor agonist cloprostenol (5 microg). Already 3 h after cloprostenol injection, FP receptor mRNA levels exhibited a pronounced increase to values 4.0-fold higher (P < 0.01) than before injection. At 7 h through 24 h, the amount of luteal FP receptor mRNA decreased, approaching preinjection levels, whereafter they were again 3.0-fold higher (P < 0.01) at 48 h than before injection. We conclude that following homologous stimulation of the FP receptor, abundance of this mRNA is tissue-specifically regulated in a dynamic pattern, suggestive of an important role for FP receptor-mediated action on gene expression during the demise of corpus luteum function.
最近几个编码前列腺素（PG）受体的cDNA的克隆为更详细地研究前列腺素在黄体功能中假定的调节作用铺平了道路。我们利用逆转录-聚合酶链反应（RT-PCR），以基于最近克隆的小鼠cDNA的寡核苷酸为引物，分离出一种编码卵巢前列腺素F2α（FP）受体的mRNA。大鼠卵巢mRNA的5'-非翻译区是通过5'-RACE（cDNA末端快速扩增）分离出来的。分离出的1526个碱基对序列跨越整个开放阅读框，发现在蛋白质编码区与从大鼠星形胶质细胞cDNA文库中分离出的类似序列100%相同，但在5'-非翻译区的前32个核苷酸不同，这可能是由于组织特异性剪接异质性。使用核糖核酸酶保护试验，在成年假孕大鼠（第8天）皮下注射黄体溶解剂量的FP受体激动剂氯前列醇（5微克）后的不同时间点（0.5-48小时），对切除的黄体中FP受体mRNA水平进行了定量分析。氯前列醇注射后仅3小时，FP受体mRNA水平就显著增加，达到比注射前高4.0倍的值（P<0.01）。在7小时至24小时期间，黄体FP受体mRNA的量减少，接近注射前水平，此后在48小时时又比注射前高3.0倍（P<0.01）。我们得出结论，在FP受体的同源刺激后，这种mRNA的丰度以动态模式进行组织特异性调节，这表明在黄体功能衰退期间，FP受体介导的基因表达作用具有重要作用。
