斑马鱼促甲状腺激素生物节律的研究

项目名称： 斑马鱼促甲状腺激素生物节律的研究

项目编号： No.31300969

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 生物科学

项目作者： 季成

作者单位： 苏州大学

项目金额： 21万元

中文摘要： 甲状腺素在维持生物体生长，发育和新陈代谢等方面起到重要的调节作用,而主导调控甲状腺活性的则是促甲状腺激素（TSH）。TSH 是由α和β亚基构成的糖蛋白复合体，其中β亚基决定TSH对其受体的特异性。申请者已克隆和鉴定了斑马鱼tshβ基因的启动子，并且发现其拥有生物钟调控元件E-Box，但是TSH节律调控的分子机制以及TSH可能对生物节律的影响尚需深入研究。本研究拟采用qPCR和整体原位杂交技术以及构建tshβ:luc转基因斑马鱼检测TSH表达的节律的特征；通过体外细胞转染手段和斑马鱼生物钟基因无效突变体(per1b-/- )检验不同生物钟基因对tshβ的调控作用；利用TALEN构建tshβ-/- 突变鱼研究tshβ对生物钟基因表达和运动节律的影响。该研究将有助于深入理解TSH的生理功能和节律调节的作用机制，在临床上对甲状腺类疾病的诊断和治疗具有重大。

中文关键词： 促甲状腺激素；斑马鱼；TALEN；生物节律；

英文摘要： Thyroid hormones (T4 and T3) produced by the thyroid gland play a critical role in animal growth, development,and metabolism. The activity of the thyroid gland is regulated predominantly by thyroid stimulating hormone (TSH) secreted by thyrotrope cells in the anterior pituitary gland. TSH is a glycoprotein complex composed of subunits α and β. It is the β subunit (TSHβ) that determines the specificity of TSH to its receptor. Previously, we characterized the zebrafish tshβ gene promoter, which harbors circadian regulation-related E-Boxes. However, molecular mechanisms underlying circadian regulation of tshβ and the effects of tshβ on expression of circadian clock genes as well as locomotor activities remain elusive. In this study, qPCR, in situ hybridization, and tshβ:luc transgenic fish lines will be employed to examine circadian expression of tshβ; in vitro cell transfection assays and a zebrafish circadian clock gene null mutant (per1b-/- ) will be used to examine the regulatory roles of circadian clock genes in tshβ expression; and tshβ-/- zebrafish generated via TALEN (transcription activator-like effector nuclease) will be used to examine the effects of tshβ on expression of circadian clock genes and locomotor rhythmicity . This study should provide invaluable insights into circadian regulation of tshβ and

英文关键词： Thyroid stimulating hormone；zebrafish；TALEN；circadian rhythms；