Mechanisms of Perchlorate-Induced Disruption of Sexual Differentiation

高氯酸盐引起的性别分化破坏的机制

• 批准号: 8399107
• 负责人: Charles Loren Buck
• 金额: $ 50.8万
• 依托单位: UNIVERSITY OF ALASKA ANCHORAGE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-01 至 2014-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8399107
• 关键词: Affect; Animal Model; Animals; Antisense Oligonucleotides; Aromatase; Behavior; Behavioral; Courtship; Development; Disease; Dose; Endocrine Disruptors; Environment; Epidemic; Family member; Female; Fishes; Food; Frequencies; Functional disorder; Gasterosteidae; Gene Expression Profile; Gene Family; Genes; Genetic; Genetic Transcription; Genome; Goals; Gonadal structure; Gonadotropins; Health; Hormonal; Human; Human Milk; Hypertrophy; In Situ Hybridization; Induced Mutation; Ingestion; Iodides; Maintenance; Mediating; Messenger RNA; Milk; Molecular; Molecular Profiling; Morphology; Mutate; Mutation; Outcome; Pathway interactions; Perchlorates; Phenotype; Physiological; Physiology; Production; Proteins; Reproductive Health; Research; Risk; Role; SLC5A5 gene; Sex Differentiation; Sexual Development; Supplementation; Testing; Testis; Thyroid Diseases; Thyroid Function Tests; Thyroid Gland; Thyroid Hormones; Tissues; Transcript; Translating; United States; Water; Work; Zinc Fingers; design; developmental disease; experience; gain of function; gene function; geographic difference; human disease; knock-down; loss of function; male; nuclease; paralogous gene; public health relevance; receptor; reproductive; research study; response; sodium-iodide symporter; toxicant; uptake

DESCRIPTION (provided by applicant): The recent dramatic increase and geographic differences in frequency of reproductive diseases are likely influenced by changes in the environment, including perchlorate exposure. Perchlorate (ClO4-) is a persistent, chlorinated water-soluble contaminant that is pervasive in the United States. As a toxicant, perchlorate poses a major risk to human health through ingestion of contaminated water, food, and breast milk. Perchlorate is a known endocrine disruptor that competitively inhibits iodide uptake at the Sodium-Iodide Symporter (NIS) in the thyroid, thus hindering thyroid hormone synthesis. Studies demonstrate, however, that perchlorate exposure masculinizes both female and male stickleback fish (Gasterosteus aculeatus), leading to hermaphroditic females and males with testicular hypertrophy, results that are not predicted by a simple, direct thyroid- disruption mechanism. The goal of this project is to reconcile the dominant paradigm of perchlorate action - exclusively by disruption of NIS in the thyroid - with masculinization of behavior, physiology, and morphology in stickleback. The project's goal is to identify previously unsuspected pathways by which perchlorate may impact human reproductive health. Our working hypothesis is that perchlorate disrupts gonadal development by acting independently of the thyroid. Aim 1 will determine whether all observed phenotypic responses to perchlorate exposure in stickleback are mediated by the thyroid by rescuing thyroid hormone levels in perchlorate-exposed fish. Aim 2 will define the functional roles of NIS and NIS-paralogs in disruption of gonadal development by perchlorate using in situ hybridization to localize mRNA (Aim 2a), loss-of-function experiments to knock down expression of NIS and NIS-paralogs with morpholino anti-sense oligonucleotides and induced mutations using zinc finger nucleases (Aim 2b), and gain-of-function experiments by over- expressing the NIS and NIS-paralogs (Aim 2c). Aim 3 will determine the mechanism by which perchlorate alters sex differentiation using whole genome transcription profiling to determine which genes are early responders to perchlorate exposure, which are likely to be downstream genes, and whether responding genes are related to thyroid or gonad development. Quantitative PCR (qPCR) and in situ hybridization will verify expression profiling results. Significance. The proposed experiments will identify molecular and physiological pathways by which perchlorate disrupts gonadal development, whether solely via NIS in the thyroid or by other mechanisms. Because perchlorate is a pervasive contaminant in the U.S., our proposed work has direct implications for human health, particularly regarding thyroid diseases and disorders of sexual development.

描述（由申请人提供）：近期的急剧增加和生殖疾病频率的地理差异可能受环境变化的影响，包括耕地暴露。高氯酸盐（Clo4-）是一种持续的氯化水溶性污染物，在美国普遍存在。作为一种有毒物质，高氯酸盐通过摄入受污染的水，食物和母乳构成对人类健康的主要风险。高氯酸盐是一种已知的内分泌破坏者，在甲状腺中竞争性抑制碘化物分类剂（NIS）的碘化物摄取，从而阻碍甲状腺激素的合成。然而，研究表明，高氯酸盐暴露使雌性和雄性棍子鱼（gasterosteus aculeatus）均导致雌雄同体的雌性和雄性雌性具有睾丸肥大，这不是由简单，直接直接的甲状腺干扰机制预测的结果。该项目的目的是调和高氯酸盐作用的主要范式 - 仅通过甲状腺中的NIS中断与行为，生理和形态的男性化。该项目的目标是确定高氯酸盐可能会影响人类生殖健康的先前未受关注的途径。我们的工作假设是，高核通过独立于甲状腺行动来破坏性腺发育。 AIM 1将确定是否通过在高氯酸盐暴露的鱼类中挽救甲状腺激素水平来介导甲状腺中的高氯酸盐暴露的所有观察到的表型反应。 AIM 2将定义NIS和NIS-核子在腹膜开发中的功能作用，用于使用原位杂交来定位mRNA（AIM 2A）（AIM 2A），功能丧失实验，以敲低NIS和NIS-Paralogs与Morpholino抗Sensensemensencens of Z的表达，并使用Morpholino Antosens-Eligonucetiess和suntications of-nis-paralogs和诱导的寡核试剂（AIM）（Aim-nis-Paralogs）（Aim-Nis-Paralogs）（以及使用Z）降低了NIS-Paralogs（以及使用Z）。通过过度表达NIS和NIS-Paralogs（AIM 2C）来获得功能的实验。 AIM 3将通过整个基因组转录分析来确定高氯酸盐改变性别分化的机制，以确定哪些基因是早期反应者对高氯酸盐暴露的反应者，这些基因可能是下游基因，以及反应基因是否与甲状腺或性腺发育有关。定量PCR（QPCR）和原位杂交将验证表达分析结果。意义。提出的实验将确定高核破坏性腺发育的分子和生理途径，无论是仅通过甲状腺中的NIS还是其他机制。由于高氯酸盐在美国是一种普遍的污染物，因此我们提议的工作对人类健康有直接影响，特别是关于甲状腺疾病和性发展的疾病。