Cell Interactions in Male Reproductive Phthalate Injury

男性生殖邻苯二甲酸盐损伤中的细胞相互作用

• 批准号: 6850819
• 负责人: KAMIN J JOHNSON
• 金额: $ 30.15万
• 依托单位: THE HAMNER INSTITUTES
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-06-25 至 2006-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6850819
• 关键词: HeLa cells; Sertoli cells; biological signal transduction; cell adhesion; cell adhesion molecules; cell cell interaction; cytotoxicity; environmental toxicology; flow cytometry; immunocytochemistry; immunoprecipitation; in situ hybridization; laboratory rat; laser capture microdissection; male reproductive system disorder; phosphorylation; phthalates; polymerase chain reaction; protein degradation; terminal nick end labeling; testis disorder; western blottings

DESCRIPTION (provided by applicant): Phthalates are ubiquitous environmental contaminants present in soft plastics and PVC-containing devices, resulting in significant human exposure through contamination of food and medical products. The recently completed study by the National Toxicology Program Center for the Evaluation of Risks to Human Reproduction noted there was "serious concern" that some human phthalate exposures are sufficiently high to adversely affect male reproductive function. Despite being the most studied testicular toxicant, the mechanism of phthalate-induced testicular injury is not known. Therefore, the long-term objective of this research is to determine the mechanism of phthalate-induced testicular injury in molecular detail. Hallmarks of phthalate testicular injury are an initial action on Sertoli cells, disruption of Sertoli cell FSH-induced cAMP generation, and the loss of Sertoli-germ cell adhesion. These historical data suggest that both signaling from G protein-coupled receptors and Sertoli-germ cell adhesion proteins are proximal targets of phthalate injury. Recently, putative Sertoli cell G protein-coupled receptors that are cell-cell adhesion proteins (flamingos) were discovered in testis, making such proteins attractive phthalate targets. The specific down-regulation of testicular flamingo mRNA during early stages of phthalate exposure confirmed that flamingo biology was compromised by phthalates. Because of these observations, this grant proposal investigates the mechanism of phthalate-induced testicular injury by examining the following working hypothesis: phthalates induce the phosphorylation, internalization and degradation of flamingo l and 2 protein leading to Sertoli-germ cell detachment and germ cell apoptosis. Three specific aims will test this hypothesis by 1) localizing flamingo 1 and 2 to specific testis cell types, 2) examining both qualitative and quantitative changes in flamingo proteins during phthalate exposure, and 3) determining if disruption of flamingo protein mimics the phenotype produced by phthalate injury.

描述（申请人提供）：邻苯二甲酸盐是无处不在的环境 软塑料和含PVC的设备中存在的污染物，导致 通过食品和医疗产品污染的大量人类暴露。 国家毒理学计划中心最近完成的研究 评估人类繁殖风险指出，有“严重关注” 某些人类的邻苯二甲酸盐暴露足够高以至于对 男性生殖功能。尽管是研究最多的睾丸毒物，但 邻苯二甲酸酯诱导的睾丸损伤的机制尚不清楚。所以， 这项研究的长期目的是确定 邻苯二甲酸酯诱导的睾丸损伤分子细节。邻苯二甲酸盐的标志 睾丸损伤是对Sertoli细胞的初始作用，破坏了Sertoli 细胞FSH诱导的营地产生，以及Sertoli-Germ细胞粘附的丧失。 这些历史数据表明，G蛋白偶联的两个信号传导 受体和Sertoli-Germ细胞粘附蛋白是近端靶标 邻苯二甲酸酯损伤。最近，推定的Sertoli细胞G蛋白偶联受体 在睾丸中发现了细胞细胞粘附蛋白（火烈鸟）， 使此类蛋白质具有吸引力的邻苯二甲酸酯靶标。特定的下调 邻苯二甲酸酯暴露的早期睾丸火烈鸟mRNA证实 邻苯二甲酸酯损害了火烈鸟的生物学。因为这些 观察结果，该赠款提案调查了 邻苯二甲酸酯引起的睾丸损伤通过检查以下工作 假设：邻苯二甲酸引发磷酸化，内在化和 火烈鸟L和2蛋白的降解导致Sertoli-Germ细胞脱离 和生殖细胞凋亡。三个具体目标将通过1来检验该假设。 将火烈鸟1和2定位于特定的睾丸细胞类型，2）检查两者 邻苯二甲酸酯期间火烈鸟蛋白的定性和定量变化 3）确定火烈鸟蛋白的破坏是否模仿 邻苯二甲酸酯损伤产生的表型。