内蒙古绒山羊毛囊发育、生长周期及相关基因的研究

【作者】 尹俊；

【导师】 扈廷茂； 李金泉；

【作者基本信息】 内蒙古大学 ， 动物学， 2004， 博士

【摘要】 山羊绒具有细软、保暖、洁白等特点，是纺织纤维中的珍品。结合形态学开展绒毛生长分子调控机理的研究，在分子水平上阐明羊绒生长调控的生理学和遗传学基础，不仅在基础理论方面有重要的意义，同时也为人工调控羊绒生长以及山羊的育种提供可靠的科学理论依据。 内蒙古绒山羊在胎儿期皮肤处于不断分化发育过程，表皮在妊娠40天时只有单层扁平细胞，妊娠75天形成完整的表皮结构。妊娠75～85天真皮逐渐分化出乳头层和网状层，妊娠120～135天结构基本稳定。内蒙古绒山羊胎儿体侧部初级毛囊原始体最早发生在妊娠55天，毛囊最初由上皮细胞分化为未成熟的初级毛囊，经分化发育包围真皮乳头，初级毛囊逐渐成熟。次级毛囊是从正在发育的初级毛囊靠近表皮的部分分支出来的，而不是从表皮直接发育的。妊娠95天毛囊内出现毛纤维，妊娠105天有粗毛长出皮肤，同时毛囊群落已经形成，初级毛囊在妊娠95～120天发育速度最快，妊娠120～140天后发育变缓；次级毛囊最早发生在妊娠65天，在妊娠115～125天见到绒毛穿出体表。形成的毛囊群主要为三元毛囊群，S／P为0.2～5.99：1。初级毛囊的发生主要集中在妊娠65～85天，次级毛囊的发生高峰大约在妊娠95～140天左右。毛囊的生长方向和毛球弯曲的方向、次级毛囊发生的部位都呈明显的极性现象。 5～8月份次级毛囊逐渐进入兴盛前期，兴盛前期的毛囊是随着旧绒从毛囊内脱出开始重建的新毛囊；8～12月份为毛囊的兴盛期；1月份大部分毛囊进入退行期，2～4月份为休止期。退行期的毛囊仍具有活性，绒生长变慢，休止期毛囊大部分失去活性，绒不再生长；兴盛期的毛囊毛乳头扁平，退行期、休止期的毛囊毛乳头呈尖状。绒长测量结果表明阿尔巴斯绒山羊最早于6月下旬绒毛长出体表，绒生长的最快时期为10月份；绒长出体表即使在同一品种内因个体不同而有差异，同一个体的不同部位也具有不同步性：绒毛先在股部长出体表，然后是背部，最后是体侧部。 以105天胚胎体侧部皮肤为实验材料，构建了cDNA质粒文库。得到ESTs 846个，平均长度为443.2bp。525己知功能基因被证明，可分为细胞分裂类18个，细胞信号65个；细胞结构蛋白基因73个；细胞防御类21个；基因／蛋白表达类126个；代谢类69个；未分类153个。发现了Wnt-4、BMPR-IB、ASIP gene、Ectodysplasin A等重要的和毛囊分化有关的候选基因。 构建了成年绒山羊次级毛囊兴盛期皮肤组织cDNA文库。在GenBank数据库注册非冗余ESTs 392条。序列号为CD051766一CD052157。共有319个己知功能基因。未分类的基因108个;基因和蛋白质表达类70个;代谢类有43个;细胞结构类45个;细胞信号和传导类30个;细胞和机体防御的基因巧个;参与细胞分裂的基因8个。发现了一些可能和绒毛生长有关的重要的候选基因:TGFp结合蛋白，EDRK富含因子、生长因子受体、EGF受体、G蛋白偶连受体、毛乳头驱动蛋白2、Homeobox基因otxZ、Kal 1 ikrein7、Jagged一l、Thrombospondin一l和p53诱导蛋白等。 在105天胚胎皮肤cDNA文库中发现毛发特异的角蛋白及关联蛋白基因20个。 在成年羊皮肤cDNA文库中发现毛发特异的角蛋白及角蛋白关联蛋白基因31个。初步研究了26个毛角蛋白及角蛋白关联蛋白全长cDNA序列的特征。其中24个全长cDNA己在GenBank数据库中注册，GenBank注册号为:AY3 10749.1一AY3 10754.1，AY316158.1，AY510121.l~AY510123.1，AY502950.1，12 AY50295 1.1，AY510117.1~AY510120.1，AY510124.l，AY510113.1~AYS 10116.1，AY510111.1，AY510112.IAY510110.1。2个I{AP4全长eDNA正在提交。山羊队P cDNAS及推导的KAPS在序列上有各自的特点。经原位杂交证明山羊I型毛发角蛋白KRTHAI InRNA在皮肤兴盛期初级和次级毛囊均有表达。IDE一LC/MS鉴定了绒毛中和其它动物同源的角蛋白8个，山羊毛发角蛋白及角蛋白关联蛋白4个。 本实验以内蒙古阿尔巴斯绒山羊为对象，研究了胚胎期皮肤毛囊发生发育、成年羊毛囊周期性变化的规律。在此基础上，分析了105天胚胎体侧部皮肤和成年羊皮肤的ESTS，发现了一些与毛囊发育和生长周期相关的重要候选基因，为下一步工作打下了很好的基础。更多还原

【Abstract】 Cashmere is one of the softest, warmest, finest material for textile = Combination the morphologic character, research on the mechanism of cashmere growth, elucidate the physiology and genetics foundation of cashmere growth on the molecular level, not only having the important meaning in the foundation theories, also in cross breeding and the artificial adjusts to control the cashmere growth provides dependable science theories.The skin is rapidly of differentiation and proliferation in the embryo period of goat. Only have a single cell layer in epidermis on 40 days after pregnancy, the complete epidermis construction form on 75 days. The dermis divides into the papilla and reticular layer gradually on 85 days after pregnancy. Skin construction is stable from 120 to 135 days after pregnancy. The placodes of primary hair follicle firstly appear on embryo midflank skin on 55 days, then the hair follicle grows down into the dermis and the dermal condensate remains as a discrete structure, or prepapilla, that maintains its position at the base of the extending follicle plug. Secondary hair follicle is derived from branches of primary hair follicle, rather than initiation from the epidermis . Hair growth from primary hair follicles on the age of 95 days, cashmere growth out of skin on the age of 105 days, cashmere growth out of skin on the age of 115 days. The follicle population was largely composed of primary trio groups, the S/P ratio was 0.2~5.99:l.The genesis of the primary hair follicle concentrates primarily on 65~85 days, and of secondary hair follicles have a high peak of the time from 95 to 140 days. The direction of hair follicles growth and location of the secondary hair follicle are obvious polarity.The hair follicles enter into pro-anagen from May to August gradually. The new pro-anagen follicles rebuild from old hair follicles. From August to December, they enter into anagen. Most of them enter into catagen in January. The period from February to April is their telagen. The follicle depth is most deep in August, the hair papilla breadth is widest also in August. Epidermis became thicker in April and decline in October. In catagen, the secondary hair follicles still is active, but the cashmeres grow slowly. In telagen, hair follicles lose their activity and the cashmeres no longer grow. The secondary follicle papillas are flat, dermal papilla cells (DPC) sinks inside in anagen, hair papilla is sharp in catagen and telagen, DPC is outward. The result show that the cashmeres grow out of skin earliest in June, they grow fast in October. The time of cashmere growing out of skin is different individually in same species, even different at different part in the same individual. Cashmeres grow out from hip firstly, then on the back of the body, finally from midflank.We constructed a cDNA library with mRNA from skin of 105 days’ embryo. Analyzed 846 ESTs, Their average length is 443.2 bps. Five hundred twenty five known genes are identified, they can be divided into cell division (18), cell signal (65); cell construction (73), cell defense (21); gene/protein expresses (126); metabolize (69) and unclassed (153). Wnt-4, BMPR-IB, ASEP gene and Ectodysplasin A are discovered that are important candidate genes for the hair follicle development.Another cDNA library was also built using RNA extracted from the skin tissue from an adult goat. 392 redundancy ESTs, they were registered in the GenBank, GenBank Accession numbers are CD051766-CD052157, and 319 known function gene are identified based on sequence similarity, among them, unidentified is 108, gene/protein expression is 70, metabolism is 43, involved in cell structure is 45, cell signaling/cell communication is 30, cell defence/immunity is 15 and cell division is 8.Some important candidate genes that possibility involved in cashmere growth are TGF binding protein, the EDRK enriched factor, growth factor receptor, G protein associated receptor, dermal papilla derived protein 2, homeobox gene otx2, kallikrein7, jagged-1, thrombospondin-1 andp53.Twen更多还原