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牛卵巢和子宫中交替生长激素受体信使核糖核酸的表达

Expression of alternate growth hormone receptor messenger RNA in ovary and uterus of cattle.

作者信息

Heap D, Collier R J, Boyd C K, Lucy M C

机构信息

Protiva, Monsanto Company, St. Louis, MO 63198, USA.

出版信息

Domest Anim Endocrinol. 1996 Sep;13(5):421-30. doi: 10.1016/0739-7240(96)00072-0.

Abstract

Growth hormone (GH) receptor mRNA is found within the corpus luteum and endometrium of cattle. However, binding sites for placental lactogen (PL) but not GH are found within these tissues. The objectives were to isolate cDNA for the GH receptor within the reproductive tissues of cattle and to examine these cDNA as potential variants of the GH receptor that bind PL. Ten cDNA clones were isolated from a bovine endometrial cDNA library with a 32P-labeled cDNA of the GH receptor extracellular domain. On the basis of restriction enzyme digestion, 2 of the 10 cDNA clones contained exon 1. The DNA sequence of these clones was determined by dideoxy nucleotide sequencing. The exon 1 DNA sequence of each clone (exon 1B) was different from the previously reported exon 1 for the bovine GH receptor cDNA isolated from liver (exon 1A). Analyses of these cDNA sequences showed that exon 1B contained significant homology with placental forms of the GH receptor found in mouse and human. Unlike the human cDNA, the bovine cDNA isolated from endometrium contained an intact third exon. Amplification of GH receptor mRNA by reverse transcriptase polymerase chain reaction, with exon 1A- and 1B-specific forward primers, showed that exon 1B was expressed in liver, corpus luteum, ovary, endometrium, and myometrium. Exon 1A was found almost exclusively in liver, and little was found in reproductive tissues. The predicted initiation of protein coding for the GH receptor was within the second exon and was not changed by the splicing of the alternate first exon. This suggests that the alternate mRNA results in the expression of intact GH receptor protein that is similar to that found within liver. Alternative promoters (1B) may control the expression of the receptor outside the liver. Furthermore, mechanisms other than the differential splicing of GH receptor protein may dictate the specificity of PL binding within the endometrium and corpus luteum.

摘要

生长激素(GH)受体mRNA存在于牛的黄体和子宫内膜中。然而,在这些组织中发现了胎盘催乳素(PL)的结合位点,却未发现GH的结合位点。研究目的是分离牛生殖组织中GH受体的cDNA,并将这些cDNA作为结合PL的GH受体潜在变体进行研究。用32P标记的GH受体胞外域cDNA从牛子宫内膜cDNA文库中分离出10个cDNA克隆。根据限制性酶切分析,10个cDNA克隆中有2个包含外显子1。通过双脱氧核苷酸测序确定了这些克隆的DNA序列。每个克隆的外显子1 DNA序列(外显子1B)与先前报道的从肝脏分离的牛GH受体cDNA的外显子1(外显子1A)不同。对这些cDNA序列的分析表明,外显子1B与在小鼠和人类中发现的胎盘形式的GH受体具有显著同源性。与人类cDNA不同,从子宫内膜分离的牛cDNA包含完整的第三个外显子。用外显子1A和1B特异性正向引物通过逆转录聚合酶链反应扩增GH受体mRNA,结果表明外显子1B在肝脏、黄体、卵巢、子宫内膜和子宫肌层中表达。外显子1A几乎只在肝脏中发现,在生殖组织中很少发现。预测的GH受体蛋白质编码起始于第二个外显子,并且不会因交替的第一个外显子的剪接而改变。这表明交替的mRNA导致完整的GH受体蛋白质表达,该蛋白质与在肝脏中发现的相似。替代启动子(1B)可能控制肝脏外受体的表达。此外,除了GH受体蛋白质的差异剪接之外的机制可能决定了子宫内膜和黄体中PL结合的特异性。

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