Mediators for dynamic regulation of Star transcription in Leydig cells

Leydig 细胞中 Star 转录动态调节的介质

基本信息

批准号：
10152639
负责人：
COLIN ROBERT JEFCOATE
金额：
$ 55.72万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-08-16 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10152639
关键词：
Acute Adult Age Androgens Animals Biology Cell Differentiation process Cell physiology Cells Cholesterol Chromatin Clinical Congenital Abnormality Cyclic AMP Cyclic AMP-Dependent Protein Kinases Data Diagnosis Disease Endocrine Endocrine Disruptors Endocrine disruption Environment Enzymes Erinaceidae Event Excision Exposure to Fatigue Fetal Development Fluorescent in Situ Hybridization Gatekeeping Gene Expression Regulation Genes Genetic Diseases Genetic Transcription Genitourinary system Health Image In Vitro Individual Infertility Kinetics Knowledge Learning Life Maintenance Mediator of activation protein Messenger RNA Microfluidics Molecular Newborn Infant Output Paracrine Communication Pathway interactions Pattern Physiologic pulse Physiological Play Population Pregnancy Production Prostate Puberty Publishing RNA RNA Splicing Regulation Reporting Resolution Role Series Sex Differentiation Disorders Signal Transduction Sleep Apnea Syndromes Source Steroids Stimulus Swelling Syndrome Techniques Technology Testing Testis Testosterone Time Trans-Activators Transcript Transcriptional Regulation arm bone loss bone mass cholesterol control design experimental study fetal genetic regulatory protein human old age (65+)in vivo innovation leydig interstitial cell male men molecular imaging mouse model muscle form novel paracrine prevent public health relevance response sertoli cell sex development disorder subfertility testosterone replacement therapy tool trend

Acute Adult Age Androgens Animals Biology Cell Differentiation process Cell physiology Cells Cholesterol Chromatin Clinical Congenital Abnormality Cyclic AMP Cyclic AMP-Dependent Protein Kinases Data Diagnosis Disease Endocrine Endocrine Disruptors Endocrine disruption Environment Enzymes Erinaceidae Event Excision Exposure to Fatigue Fetal Development Fluorescent in Situ Hybridization Gatekeeping Gene Expression Regulation Genes Genetic Diseases Genetic Transcription Genitourinary system Health Image In Vitro Individual Infertility Kinetics Knowledge Learning Life Maintenance Mediator of activation protein Messenger RNA Microfluidics Molecular Newborn Infant Output Paracrine Communication Pathway interactions Pattern Physiologic pulse Physiological Play Population Pregnancy Production Prostate Puberty Publishing RNA RNA Splicing Regulation Reporting Resolution Role Series Sex Differentiation Disorders Signal Transduction Sleep Apnea Syndromes Source Steroids Stimulus Swelling Syndrome Techniques Technology Testing Testis Testosterone Time Trans-Activators Transcript Transcriptional Regulation arm bone loss bone mass cholesterol control design experimental study fetal genetic regulatory protein human old age (65+)in vivo innovation leydig interstitial cell male men molecular imaging mouse model muscle form novel paracrine prevent public health relevance response sertoli cell sex development disorder subfertility testosterone replacement therapy tool trend

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项目摘要

7. Project summary/Abstract Leydig cells within the testis are the source of androgens that promote virility at both fetal and adult stages, but Leydig cell populations are distinct at each age. As such, adult and fetal Leydig cells function to synthesize testosterone in distinct cellular and endocrine environments. The AIMs of this proposal are focused on regulation of Star, the known gatekeeper in controlling access of cholesterol to enzymatic activity of the series of steroidogenic enzymes required for conversion to testosterone. Here we focus on novel regulatory events that exert dynamic interactions with Star chromosomal loci that will explain how fluctuating Star transcript accumulation can relate to changes in androgen synthesis. Previously, we used high-resolution fluorescent in situ hybridization (HR-FISH) to localize and quantify a unique pattern for primary, spliced, and mRNA species accumulation for Star compared to other steroidogenic genes within single Leydig cells. We will use this and other innovative techniques to compare results from studies that investigate individual adult and fetal Leydig cells in vitro, within MA10 cells and primary cultures, and in vivo, within whole testes. Pulsatile LH stimulates primarily cAMP/PKA signals to promote testosterone synthesis in adult Leydig cells. While we have substantial means to explain how Star transcription is turned ON by LH/PKA, we understand little about what happens when the pulse is removed, and even less about the interpulse interval. In AIM 1, we will test the hypothesis that the events occurring at Star loci during this interval are just as critical to controlling testosterone output as the initial stimulus. Meanwhile, the external stimuli that maintain androgen synthesis in fetal Leydig cells are less clear, but evidence points to paracrine signals, with PKA activity playing a role. Another paracrine factor, Sertoli cell-derived Desert Hedgehog (Hh) is known to initiate fetal Leydig cell differentiation, but its role in their maintenance has not been tested. Once differentiated, fetal Leydig cells produce androgens at a steadily increasing rate until late gestation. Therefore, in AIM 2, we will test the hypothesis that regulatory events on Star loci facilitate a controlled increase in androgens within the fetal Leydig cell that compare to those that occur during the interpulse interval in adult Leydig cells. Our findings have the potential to explain fundamental biology underlying steroidogenic control and will have a profound impact on our ability to explore mechanisms by which disturbances in testosterone synthesis, as in endocrine disruption, cause significant clinical ramifications in males from all stages of life.

7。项目摘要/摘要睾丸中的leydig细胞是雄激素的来源阶段，但Leydig细胞种群在每个年龄段是不同的。因此，成年和胎儿leydig细胞起作用在不同的细胞和内分泌环境中合成睾丸激素。该提议的目的是专注于恒星的调节，这是已知的守门人，用于控制胆固醇酶促的酶促转化为睾丸激素所需的一系列类固醇生成酶的活性。在这里我们关注与星染色体基因座发挥动态相互作用的新型调节事件，该事件将解释如何解释恒星转录本的波动可能与雄激素合成的变化有关。以前，我们使用的高分辨率荧光原位杂交（HR-FISH）来定位和量化独特的模式与其他类固醇基因相比，对于恒星的一级，剪接和mRNA物种的积累在单个Leydig细胞中。我们将使用此和其他创新技术来比较研究在MA10细胞内研究的成年和胎儿Leydig细胞的研究培养物和体内整个睾丸。脉冲LH主要刺激营地/PKA信号以促进成年leydig细胞中的睾丸激素合成。虽然我们有实质性的方法来解释如何明星转录是由LH/PKA打开的，我们对脉冲是什么几乎了解删除，甚至更少关于插槽间隔。在AIM 1中，我们将测试事件的假设在此时间间隔内发生在Star基因座，对于控制睾丸激素的输出与初始相同刺激。同时，在胎儿leydig细胞中维持雄激素合成的外部刺激较少清楚，但证据表明旁分泌信号，PKA活动起着作用。另一个旁分泌因子，已知Sertoli细胞来源的沙漠刺猬（HH）引发胎儿Leydig细胞分化，但其作用在他们的维护中尚未进行测试。一旦分化，胎儿leydig细胞在A处产生雄激素稳步提高速率，直到妊娠晚期。因此，在AIM 2中，我们将测试调节性的假设星座上的事件可有助于受控的胎儿Leydig细胞中的雄激素增加对于成年leydig细胞间间隔内发生的情况。我们的发现有可能解释基本的生物学基础类固醇生成控制，并将对我们的我们产生深远的影响能够探索睾丸激素合成中干扰的机制，例如内分泌破坏，从生活的各个阶段引起男性的重大临床影响。