TARGETS OF THYROID HORMONE ACTION IN DEVELOPING BRAIN

甲状腺激素在大脑发育中的作用目标

• 批准号: 2145283
• 负责人: CATHERINE C THOMPSON
• 金额: $ 6.1万
• 依托单位: CARNEGIE INSTITUTION OF WASHINGTON, D.C.
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-03-01 至 2000-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2145283
• 关键词: RNase protection assay; animal genetic material tag; antiserum; complementary DNA; computer assisted sequence analysis; developmental genetics; developmental neurobiology; gene expression; genetic library; genetic regulation; hormone regulation /control mechanism; human genetic material tag; immunocytochemistry; in situ hybridization; laboratory rabbit; laboratory rat; molecular cloning; newborn animals; northern blottings; nucleic acid probes; nucleic acid sequence; protein sequence; southern blotting; triiodothyronine; western blottings

The overall objective of this project is to gain an understanding of the molecular mechanisms underlying the development of the mammalian central nervous system. Among the many physiological factors that influence the central nervous system, thyroid hormone is known to be essential for proper development. Thyroid hormone deficiency during a short critical period produces severe neurological defects in humans and in experimental animals. The actions of thyroid hormone are mediated through nuclear receptors, which regulate the expression of specific target genes in response to hormone binding. Although much is known about the function of thyroid hormone receptors, the genes regulated by these receptors in the developing brain are largely unknown. The main objective of the research proposed here is to identify and characterize genes whose expression in the brain is regulated by thyroid hormone during the period of time in which thyroid hormone is known to be critical for proper development. Potential target genes have been identified using subtractive hybridization coupled with polymerase chain reaction amplification to isolate genes that are up-regulated in response to thyroid hormone in developing rat brain. Developmental and tissue specific patterns of expression for the candidate genes will be determined,using Northern analysis and in situ hybridization to identify those most likely to have a role in central nervous system development. The complete nucleotide sequence will be determined for selected genes and the primary amino acid sequence deduced. Antibodies will be produced to determine expression patterns and subcellular localization of the proteins using Western analysis and immunohistochemical methods. An important long term goal of this project is to determine the functional role of thyroid hormone responsive gene products in nervous system development. A detailed understanding the mechanisms by which thyroid hormone effects brain development should be of value in ultimately attempting to treat developmental and degenerative diseases of the central nervous system.

该项目的总体目标是了解 哺乳动物中枢发育的分子机制 神经系统。在众多影响生理因素的因素中 众所周知，甲状腺激素对中枢神经系统至关重要 适当的发展。短暂危急期间甲状腺激素缺乏 时期会在人类和实验中产生严重的神经缺陷 动物。甲状腺激素的作用是通过核介导的 受体，调节特定靶基因的表达 对激素结合的反应。尽管人们对它的功能了解很多 甲状腺激素受体，受这些受体调节的基因 发育中的大脑在很大程度上是未知的。研究的主要目标 这里提出的是识别和表征其表达的基因 大脑在一段时间内受到甲状腺激素的调节 已知哪种甲状腺激素对于正常发育至关重要。 潜在的靶基因已通过消减法鉴定出来 杂交与聚合酶链式反应扩增相结合 分离出响应甲状腺激素而上调的基因 正在发育的老鼠大脑。发育和组织特异性模式 将使用 Northern 确定候选基因的表达 分析和原位杂交以确定那些最有可能具有 在中枢神经系统发育中发挥作用。完整的核苷酸 将确定所选基因和主要氨基酸的序列 推导出来的序列。将产生抗体以确定表达 使用 Western 分析蛋白质的模式和亚细胞定位 分析和免疫组织化学方法。一个重要的长期目标 该项目旨在确定甲状腺激素的功能作用 神经系统发育中的反应性基因产物。详细的 了解甲状腺激素影响大脑的机制 发展应该对最终尝试治疗具有价值 中枢神经系统的发育和退行性疾病。