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人胎盘中pit-1信使核糖核酸和蛋白质的表达

Expression of pit-1 messenger ribonucleic acid and protein in the human placenta.

作者信息

Bamberger A M, Bamberger C M, Pu L P, Puy L A, Loh Y P, Asa S L

机构信息

Laboratory of Developmental Neurobiology, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892, USA.

出版信息

J Clin Endocrinol Metab. 1995 Jul;80(7):2021-6. doi: 10.1210/jcem.80.7.7608249.

Abstract

It is well established that the human placenta produces a wide range of hormones similar to those secreted by the pituitary and hypothalamus. However, the physiological role and regulation of placental hormone synthesis and release are still largely unknown. GH (GH-N) is expressed in the pituitary, where it requires the tissue-specific transcription factor Pit-1. Chorionic somatomammotropin A (CS-A) and CS-B as well as the placental GH variant (GH-V), which also belong to the GH gene family and are located in the same chromosomal cluster, are expressed in the placental syncytiotrophoblast. The presence of Pit-1-binding sites in the CS-A and GH-V promoter regions predicts that Pit-1 may be expressed in the placenta. However, this has not yet been demonstrated. To examine possible similarities in the regulation of these genes in the pituitary and placenta, we studied the expression of pit-1 messenger ribonucleic acid (mRNA) in the human placenta, transformed human placental cells, and the JEG-3 choriocarcinoma cell line. Polymerase chain reaction (PCR) products of the expected size were amplified from first and third trimester placentas, transformed placental cells, and JEG-3 complementary DNA by reverse transcription-PCR. The pit-1-specific sequence was confirmed by restriction endonuclease digestion, Southern hybridization, and DNA sequencing. Human pituitary tissue was used as a positive control; no PCR product was obtained from hippocampus (negative control). In situ hybridization of placental tissue sections revealed the presence of pit-1 mRNA in first and third trimester syncytiotrophoblast. Pit-1 protein was localized by immunohistochemistry with the same tissue distribution and a nuclear localization pattern. These data demonstrate expression of pit-1 mRNA and Pit-1 protein in the human placenta, thus questioning its role as a pituitary-specific regulator of GH-N gene transcription. The expression of Pit-1 in the placenta, together with its previously demonstrated capability to bind to and activate the CS-A and the GH-V promoters, suggests that it may play a role in the regulation of hormones belonging to the GH gene family in both pituitary and placenta.

摘要

众所周知,人类胎盘会产生多种与垂体和下丘脑分泌的激素相似的激素。然而,胎盘激素合成与释放的生理作用及调节机制在很大程度上仍不为人知。生长激素(GH-N)在垂体中表达,在垂体中它需要组织特异性转录因子Pit-1。绒毛膜生长催乳素A(CS-A)和CS-B以及同样属于GH基因家族且位于同一染色体簇的胎盘生长激素变体(GH-V)在胎盘合体滋养层中表达。CS-A和GH-V启动子区域中存在Pit-1结合位点,这预示着Pit-1可能在胎盘中表达。然而,这一点尚未得到证实。为了研究垂体和胎盘中这些基因调节的可能相似性,我们研究了人胎盘、转化的人胎盘细胞和JEG-3绒癌细胞系中pit-1信使核糖核酸(mRNA)的表达。通过逆转录聚合酶链反应(PCR)从妊娠早期和晚期胎盘、转化的胎盘细胞以及JEG-3互补DNA中扩增出预期大小的PCR产物。通过限制性内切酶消化、Southern杂交和DNA测序确认了pit-1特异性序列。人垂体组织用作阳性对照;未从海马体中获得PCR产物(阴性对照)。胎盘组织切片的原位杂交显示在妊娠早期和晚期合体滋养层中存在pit-1 mRNA。通过免疫组织化学将Pit-1蛋白定位,其具有相同的组织分布和核定位模式。这些数据证明了pit-1 mRNA和Pit-1蛋白在人胎盘中的表达,从而对其作为GH-N基因转录的垂体特异性调节因子的作用提出了质疑。Pit-1在胎盘中的表达,连同其先前证明的与CS-A和GH-V启动子结合并激活的能力,表明它可能在垂体和胎盘中对属于GH基因家族的激素调节中发挥作用。

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