Berry S A, Srivastava C H, Rubin L R, Phipps W R, Pescovitz O H
Department of Pediatrics, University of Minnesota, Minneapolis 55455.
J Clin Endocrinol Metab. 1992 Jul;75(1):281-4. doi: 10.1210/jcem.75.1.1619020.
Although the sequence of human GH-releasing hormone (GHRH) has been determined, all of the information concerning gene expression has been based on pathological sources of ectopic GHRH, since the only established physiological source of GHRH in humans is the hypothalamus. We recently reported the presence of extrahypothalamic GHRH-like mRNA and immunoreactive material in rat testis and placenta. To determine if human testis and placenta also contain immunoreactive GHRH-like peptides, tissue extracts were analyzed using enzyme-linked immunosorbent assay methodology. Both tissues had detectable quantities of immunoreactive peptide recognized by a monoclonal antibody to synthetic human GHRH-(1-44) (testis, 2.4 ng/g tissue, 0.68 ng/mg protein; placenta, 2.6 ng/g tissue, 0.36 ng/mg protein). The origin of these peptides was confirmed by extracting total RNA from human testis and placenta, with analysis on Northern blots probed with riboprobes for rat hypothalamic GHRH cDNA and human pancreatic tumor GHRH cDNA. Human testis and placenta total RNA both contain an approximately 790-nucleotide RNA species similar in size to that reported in ectopic GHRH-producing human tumors. In addition, two larger hybridization signals were seen at 3000 and 4900 nucleotides. These data suggest that testis and placenta are extrahypothalamic sites of expression of the human GHRH gene. Normal expression of the GHRH gene in extrahypothalamic sites may include transcription of larger mRNA species than those observed in ectopic pathological sources of GHRH expression.
虽然人类生长激素释放激素(GHRH)的序列已被确定,但所有关于基因表达的信息都基于异位GHRH的病理来源,因为人类中唯一已确定的GHRH生理来源是下丘脑。我们最近报道在大鼠睾丸和胎盘中存在下丘脑外的GHRH样mRNA和免疫反应性物质。为了确定人类睾丸和胎盘是否也含有免疫反应性GHRH样肽,使用酶联免疫吸附测定方法对组织提取物进行了分析。两种组织都含有可检测量的免疫反应性肽,该肽可被针对合成人GHRH-(1-44)的单克隆抗体识别(睾丸,2.4 ng/g组织,0.68 ng/mg蛋白质;胎盘,2.6 ng/g组织,0.36 ng/mg蛋白质)。通过从人类睾丸和胎盘中提取总RNA,并使用大鼠下丘脑GHRH cDNA和人类胰腺肿瘤GHRH cDNA的核糖探针进行Northern印迹分析,证实了这些肽的来源。人类睾丸和胎盘的总RNA都含有一种大小约为790个核苷酸的RNA物种,其大小与在产生异位GHRH的人类肿瘤中报道的相似。此外,在3000和4900个核苷酸处还观察到两个较大的杂交信号。这些数据表明睾丸和胎盘是人类GHRH基因在下丘脑外的表达部位。GHRH基因在下丘脑外部位的正常表达可能包括转录比在GHRH表达的异位病理来源中观察到的更大的mRNA物种。