REGULATION OF POMC GENE IN TESTIS AND EPIDIDYMIS

POMC基因在睾丸和附睾中的调控

• 批准号: 3232802
• 负责人: CHING-LING C CHEN
• 金额: $ 10.87万
• 依托单位: POPULATION COUNCIL
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-09-01 至 1988-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3232802
• 关键词: adrenocorticotropic hormone; epididymis; gel electrophoresis; gene expression; genetic regulation; genetic transcription; gonadotropin releasing factor; histochemistry /cytochemistry; hormone biosynthesis; hormone regulation /control mechanism; melanocyte stimulating hormone; molecular cloning; nucleic acid sequence; prolactin; testis

The objective of our research is to understand how a specific gene is expressed and regulated in male reproductive tissues. In this proposal, proopiomelanocortin (POMC) will be used as a model system. POMC, which is a precursor protein for ACTH, beta-endorphin and melanocyte-stimulating hormones, has been well characterized in many tissues. Our recent demonstration of the existence of POMC-like mRNA in testis and epididymis suggests the expression of a POMC gene and synthesis of POMC peptides in these tissues. Since the regulation of a specific gene expression in Leydig cells or epididymis has not yet been well understood, we have chosen to investigate the regulation of POMC gene expression in these tissues. Our first specific aim is to investigate both in vivo and in vitro the regulation of POMC mRNA level and POMC peptide synthesis, processing, and secretion in testis and epididymis by using factors known to affect functions of Leydig cells (gonadotropins, prolactin LHRH analogs, etc.) or epididymis (androgen, testis fluid). The regulation of POMC mRNA and its peptide levels will also be examined at cellular levels by using in situ cDNA hybridization histochemistry technique. Our second specific aim is to understand the molecular mechanisms involved in the change of POMC mRNA level by modulators. To answer this question, we will determine whether the rate of synthesis of POMC mRNA as measured by nuclear transcriptional activity of the POMC gene or the stability of POMC mRNA will be altered by modulators. Finally, the structure of heterogeneous POMC mRNAs expressed in the testis and epididymis will be characterized. The POMC-like mRNAs found in the testis, epididymis, or Leydig cell lines are approximately 150-200 bases shorter than those found in the pituitary or hypothalamus. To understand the mechanism involved in the production of heterogeneous POMC mRNA in different tissues, we will determine the POMC mRNA structure by cDNA cloning and DNA sequencing analysis, characterize POMC gene(s) isolated from rat genomic libraries, and identify the transcription unit of POMC gene and the pathway of processing nuclear precursor of POMC mRNA. The proposed research will provide useful information in studying the regulation of a specific gene expression in Leydig cells and epididymis. In addition, since POMC is a precursor for several peptide hormones and neurotransmitters, the proposed research will facilitate our understanding of the biological role of these individual peptides derived from POMC in the male reproductive physiology.

我们研究的目的是了解特定基因的方式 在男性生殖组织中表达和调节。 在此提案中， proOpiomelanocortin（POMC）将用作模型系统。 POMC，就是 ACTH，β-内啡肽和黑素细胞刺激的前体蛋白 激素在许多组织中都得到了很好的特征。 我们最近 证明睾丸和附加症中POMC样mRNA的存在 提出pOMC基因的表达和pOMC肽在 这些组织。 由于调节特定基因表达 Leydig细胞或附睾尚未得到充分了解，我们选择了 研究这些组织中POMC基因表达的调节。 我们的第一个具体目的是研究体内和体外 调节POMC mRNA水平和POMC肽合成，加工和 通过使用已知影响的因素，睾丸和附生膜分泌 Leydig细胞（促性腺激素，催乳素LHRH类似物等）的功能或 附睾（雄激素，睾丸液）。 POMC mRNA及其的调节 还将通过原位在细胞水平上检查肽水平 cDNA杂交组织化学技术。 我们的第二个具体目标是 了解与POMC mRNA变化有关的分子机制 调制器的水平。 要回答这个问题，我们将确定是否 通过核转录测量的POMC mRNA的合成速率 POMC基因的活性或POMC mRNA的稳定性将通过 调节器。 最后，表达的异质性POMC mRNA的结构 将在睾丸和附睾中进行表征。 像pomc的mRNA 在睾丸，附睾或leydig细胞系中发现 比垂体或下丘脑中的150-200底座短。 了解生产异质的机制 POMC mRNA在不同组织中，我们将确定POMC mRNA结构 通过cDNA克隆和DNA测序分析，表征POMC基因（S） 从大鼠基因组文库中分离出来，并确定 POMC基因和处理POMC mRNA的核前体的途径。 拟议的研究将为研究 调节leydig细胞和附睾中特定基因表达。 另外，由于POMC是几种肽激素的前体，并且 神经递质，拟议的研究将有助于我们的理解 这些在POMC中得出的单个肽的生物学作用 男性生殖生理学。