角蛋白IFs、KAPs基因对和田羊次级毛囊密度的影响及雄激素对其调节作用研究

Hetian sheep is on the list of the Country Protective Species of Livestock and Poultry. Their number occupy 70.97 % of the local sheep of Hetian region in southern Xinjiang. The Hetian sheep wool are coarse wool, and the best raw material of the famous “Hetian carpet”, which is one of the three treasures on the Silk Road. However, the wool follicle density of the Hetian sheep was far more lower than that of fine wool sheep’s. For improving the wool quantity, increasing the secondary follicle / primary follicle ratio (S/P ratio) of the Hetian sheep would be a main pathway. For keratin intermediate filament (KIF), keratin associated protein (KAP) are main constituents of follicle structure, it’s an important foundation to reveal the law of follicle development from the research about the KIF and KAP families genes. Many technologies such as histological section, gene clone and expression, real-time PCR, mice hybridoma cell fusion, microdissection, frozen section, immunofluorescence histochemistry would be emploied to analysis the follicle density of the Hetian sheep, quality and quantitive the follicle KIF and KAP families genes, made the polyclone and monoclone antibodies of these genes, display the expression position of them in the wool follicle, set up a model of sheep follicle culture in vitro , understand the rule of follicle cell regenration and apoptosis. If the androgen were possible to regulate these genes, the S/P ratio would be influenced by the changes of these genes. The research about wool follicle KIF、KAP families genes of Hetian sheep and the regulation of androgen to them, would be play an important role for improve the follicle quantity and wool quality.

和田羊已被列入国家级畜禽遗传资源保护名录，占新疆和田地区绵羊存栏总数的70.97%，其羊毛为异质毛，是著名丝路三宝之一“和田地毯”的最佳原料。但和田羊毛囊密度远远低于细毛羊，提高和田羊次级毛囊与初级毛囊的比率（S/P比率），是增加产毛量的重要途径。角蛋白中间丝（KIF）、角蛋白关联蛋白（KAP）是构成毛囊的主要结构组分，对KIF、KAP家族基因的研究是揭示毛囊发育规律的重要基础。利用组织切片、基因克隆与表达、实时荧光定量PCR、小鼠杂交瘤细胞融合、显微解剖、冰冻切片、免疫荧光组织化学等技术，解析和田羊毛囊密度参数；定性、定量研究毛囊的KIF、KAP家族基因；制备多克隆或单克隆抗体，研究其表达部位；建立体外培养模型，了解毛囊细胞体外再生与凋亡的规律，观察雄激素是否能够调节这些基因的表达从而影响S/P比率。研究和田羊毛囊角蛋白基因及调节激素，对提高单位绵羊的产毛量、改善羊毛品质具有重大意义。

研究内容：关于新疆南疆地方和田羊（山区型与平原型）毛囊发育相关角蛋白关联蛋白(KAP)和角蛋白中间丝(KIF)基因分析，雄激素对毛囊发育的影响，以及毛囊干细胞分化迁移规律研究。.重要结果、关键数据：.1、完成了和田羊毛囊KAP、KIF的10个基因的研究：重点发表了和田羊与卡拉库尔羊毛囊KIF1.2、KAP7基因的定量分析与真核表达研究，和田羊与卡拉库尔羊毛囊KAP8基因的定量分析与真核表达研究待发表，平原型和田羊毛囊KAP4.2基因的克隆与原核表达研究，山区型和田羊毛囊HGT蛋白 KAP6.1、KAP7与KAP8 的基因克隆分析等。.2、完成了和田羊毛囊KAP、KIF基因的表达差异、表达部位、抗体制备研究：平原型与山区型和田羊皮肤毛囊形态学观察及其KAPs、KIFs角蛋白基因的转录水平差异分析（待发表），发表了新疆南疆3个地方品种KAP7基因的多克隆抗体制备研究。.3、完成了毛囊干细胞的初步研究：发表了胎鼠皮肤无瘢痕愈合培养液优化及皮肤细胞增殖活跃区分析，EdU体外标记毛囊外根鞘干细胞的实验研究，C57脱毛小鼠与正常小鼠毛囊形态学观察及AP-1表达的差异，KM乳鼠背部皮肤毛囊发育及Sox2表达，小鼠毛囊发育中细胞增殖区及干细胞巢的形成演化，小鼠尾断端愈合形态学观察及细胞增殖活跃区分析，皮肤创伤愈合过程中短暂库增细胞增生及皮肤干细胞迁移，Remodeling 3D morphology of neonatal hair follicle by in vitro culturing of hair follicle outer root sheath（SCI收录），小鼠触须毛囊外根鞘体外再生模型的建立及干细胞迁移特征分析（SCI已录用）.4、完成了雄激素对毛囊发育影响的初步研究：初步探究雄激素对和田羊毛囊及角蛋白的影响研究（SCI已录用）；完成了双氢睾酮对小鼠毛囊外根鞘体外培养再生的影响研究（获第三届全国大学生生命科学大赛一等奖）。.5、超额完成人才培养、论文发表任务：培养研究生5人；发表论文12篇，其中SCI收录1篇，北大中文核刊9篇，科技核心期刊1篇；已录用SCI论文2篇。. 科学意义：获得了新疆南疆地方品种绵羊毛囊结构、毛囊发育相关基因、毛囊干细胞迁移分化基础数据，并将科技成果应用于南疆偏远地区。