纤毛病影响肾脏、骨骼、肌肉、中枢神经、心血管、生殖、呼吸道、感官、内脏与代谢的功能，危害人类健康，发病机理亟待探究。 中心粒蛋白Talpid3丧失引起纤毛缺失、Shh信号通路紊乱、胚胎发育异常、多囊肿肾病、内脏反位、不育和Joubert综合症。申请人发现talpid3与转运Shh通路下游转录因子Gli2/3的驱动蛋白kif7协同影响Shh信号传导, 机制不明。本项目拟研究Talpid3与Shh信号通路关键分子Glis/Sufu等的交互作用，构建其影响Shh通路的分子网络； 通过FLAG与抗体亲和/生物素-亲和素系统进行高通量蛋白复合体的纯化，研究胚胎发育中Talpid3在中心粒组配的功能以及与各信号通路转导元件的交互作用； 通过比较突变体与野生型的转录组来弄清talpid3和纤毛在幼鱼发育过程中的分子调控网络。该项目将阐释中心粒调控信号通路的机制，为诊治人类纤毛疾病提供新的科学依据和思路。

Ciliopathies affect the function of kidney, skeleton, muscle, central neural system, cardiovascular system, fertility, respiratory tract, viscera, metabolism, olfactory and visual senses. Puzzles are unsolved in the pathogenic and molecular mechanisms. Loss of function of talpid3(ta3), which encodes the centrosomal protein, causes failure of ciliogenesis, the bifurcated disruption of Shh signal transduction, the abnormal embryonic development, polycistic kidney disease, situs inversus, male infertility and Joubert Syndrome. The applicant previously discovered that Shh pathway is regulated by the synergistic effect between ta3 and the kif7, which encodes a kinesin-like protein to transport the downstream effectors Gli2/3 along the axenome to the tip of primary cilium. To understand the mechanism, our project will investigate in depth the physical interaction between Talpid3 and the key components of Shh pathway, Kif7, Gli2/3 and Sufu et al., and map the role of Talpid3 in the network of Shh pathway. By an in vivo biotinylation strategy combined with Flag and antibody based approaches for large scale affinity purification of protein complexes in zebrafish embryos, we aim to identify the role of Talpid3 in centriole assembly and the interactions with the components in other signaling pathways. By comparing the transcriptome of ta3-/- with that of wildtype, we aim to understand the molecular network regulated by the cilia at zebrafish larvae stage. Our work will elucidate the regulatory mechanism of centriole on signaling and propel the diagnosis and therapy of ciliopathies.