褪黑激素影响绒山羊绒毛生长脱落相关基因的筛选及其作用机制研究

Inner Mongolia Cashmere goat is one of the unique genetic resources in China. In this project, effects of melatonin (MT) implanting on the growth and shedding of cashmere, and the associated genes are to be investigated, by observing growth of the skin and hair follicle after MT implanting using tissue slices technique, referring to Cashmere growth. In order to explore the molecular mechanism of MT affecting the growth and shedding of cashmere, sequencing on transcription level (RNA-seq) will be used to build mRNA expression library of hair and skin follicle; and differentially-expressed-gene-based biological network will be constructed to predict more candidate genes associated with cashmere growth and shedding, by scanning and evaluating each candidate gene's characterization and reliability. In addition, mRNA and protein dynamic expression changes of these candidate genes both in cashmere goat from implanting MT group and control group will be tested, by in situ hybridization, fluorescence quantitative real-time PCR and Western Blot. This project will lay the molecular theory foundation of MT regulating the cashmere growth and shedding.

本项目以我国特有遗传资源内蒙古绒山羊为研究对象，开展受埋植褪黑激素（Melatonin, MT）影响的绒毛生长脱落相关基因的筛选鉴定及其分子调控作用机制的研究，揭示MT影响绒毛生长脱落的分子机制。拟应用组织切片技术，结合绒毛生长情况，观察埋植MT导致的绒山羊皮肤毛囊变化情况；利用转录组测序建立皮肤毛囊 mRNA表达文库，构建一个以差异表达基因为主的绒毛生长脱落相关基因功能网络，通过对表征和可靠性的评价，进一步筛选出受MT影响的绒毛生长脱落相关候选功能基因；运用原位杂交、荧光定量 PCR 和Western Blot等技术手段，检测候选功能基因mRNA、蛋白质在埋植组和对照组绒山羊皮肤毛囊上的时空表达变化情况。研究结果旨在为MT调控绒毛生长脱落的研究奠定分子理论基础。

本项目以我国特有遗传资源内蒙古绒山羊为研究对象，开展受埋植褪黑激素（Melatonin, MT）影响的绒毛生长脱落相关基因的筛选鉴定及其分子调控作用机制的研究。利用转录组测序建立了皮肤毛囊 mRNA表达文库，构建了一个以差异表达基因为主的绒毛生长脱落相关基因功能网络，通过对表征和可靠性的评价，进一步筛选出了受MT影响的绒毛生长脱落相关候选功能基因：FZD10、LEF1、NKD1、WNT11、BAMBI、PLAT、C1QB、F13A1、SERPING1、C1S、C1QC、GOAT_ENSBATP00000048849-D2、SNRNP48 等13个基因。其中，ENSBTAP0000048849-D2基因是皮肤组织中MT作用的一个潜在靶点，外源MT增加其表达量，完全改变了与其它基因相互作用的关系。7月、8月是皮肤毛囊发育的重要转折点，Jak-STAT signaling pathway 、 Complement and coagulation cascades和Wnt等信号通路对皮肤毛囊的发育具有重要的调控作用。4月、9月是MT影响皮肤毛囊发育的关键时期，Wnt信号通路，Jak-STAT 信号通路是MT影响皮肤毛囊周期性变化的关键信号通路。研究结果为MT调控绒毛生长脱落的研究奠定了分子理论基础。