TARGETS OF THYROID HORMONE ACTION IN DEVELOPING BRAIN

这个项目的总体目标是了解 哺乳动物中枢神经系统发育的分子机制 神经系统。在众多影响睡眠质量的生理因素中 中枢神经系统，已知甲状腺激素对 适当发展。甲状腺激素缺乏在短期危急状态下 月经期在人类和实验中会产生严重的神经缺陷 动物。甲状腺激素的作用是通过核调节的 受体，它调节特定的靶基因的表达 对激素结合的反应。尽管我们已经知道了很多关于 甲状腺激素受体，这些受体调控的基因在 发育中的大脑在很大程度上是未知的。这项研究的主要目的 在这里提出的是识别和表征其在 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正常发育至关重要。 利用消减技术鉴定了潜在的靶基因 杂交结合聚合酶链式反应扩增 分离对甲状腺激素反应上调的基因 发育中的大鼠脑。发育和组织特异性模式 候选基因的表达将使用Northern来确定 分析和原位杂交以确定最有可能患有 在中枢神经系统发育中的作用。完整的核苷酸 将确定选定基因和主要氨基酸的序列 推导出序列。将产生抗体来确定表达 蛋白质的蛋白质图谱和亚细胞定位 分析和免疫组织化学方法。一个重要的长期目标是 这个项目是为了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。一个详细的 了解甲状腺激素影响大脑的机制 发展应该是有价值的，最终试图治疗 中枢神经系统的发育性和退行性疾病。

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.