Wnt10b、β-catenin、FGF18基因在‘甘肃高山细毛羊’胎儿皮肤毛囊中的表达规律研究

摘要

以81~147 d胎龄的‘甘肃高山细毛羊’胚胎体侧部皮肤作为研究材料,通过冰冻切片、HE染色、荧光定量P C R、免疫组化技术研究Wnt10b、β-catenin、FGF18基因在次级毛囊形态发生过程中的表达规律.结果表明：从次级毛囊形态发生诱导期到器官形成阶段(胎龄87~108 d)Wnt10 b、FGF18基因表达量逐渐升高,在胎龄第108天Wnt10b、FGF18基因相对表达量分别达到(43.652±0.425)(13.67±0.207),在细胞分化期表达量逐渐下降；而β-catenin基因的表达量始终维持较低水平,相对表达量仅为(0.58±015)；Wnt10b 基因分别与β-catenin、FGF18基因表达水平呈显著正相关(r=0.85,P0.05).免疫组化结果显示在次级毛囊形态发生过程中Wnt10b、β-catenin 基因主要在表皮、基板、毛芽、毛钉和成熟毛囊的内根鞘、毛干部表达；而FGF18不仅在表皮、基板、毛芽、毛钉和成熟毛囊的内根鞘、毛干部表达,而且在真皮中表达.本研究初步表明,Wnt10b、β-catenin、FGF18基因通过Wnt/β-catenin 基因经典信号通路参与调控‘甘肃高山细毛羊’次级毛囊形态发生和再分化过程.%The expression patterns of Wnt10b,β-catenin and FGF18 genes during the morphogenesis of secondary follicles (SF)in fetal hair follicles of Gansu alpine fine-wool sheep were analyzed in the present study.The midside skins of the embryos of Gansu alpine fine-wool sheep with a gestational age (GA)of 81~147 d were used as the study objects.The expression patterns of Wnt10b,β-catenin and FGF18 genes during the morphogenesis of SF were studied through frozen sections,hematoxylin and eosin (H.E)stai-ning,real-time quantitative polymerase chain reaction (PCR)and immunohistochemical techniques.The re-sults showed that the morphogenesis of the secondary-original follicles(SO)of the Gansu alpine fine-wool sheep underwent 3 periods and 8 steps before the follicles matured.The original SO could further differen-tiate into secondary-derived follicles (SD).The gene expression levels of Wnt10b,β-catenin and FGF18 gradually increased from the induction phase of the SF morphogenesis to the organogenesis period (GA 87-108 d).At GA 108 d,the relative expression levels of Wnt10b and FGF18 reached (43.652±0.425)and (13.67±0.207),respectively,and then gradually decreased during the cell differentiation period.However, the gene expression ofβ-catenin remained at a relatively low level,while its relative expression level was only (0.58±015).The correlations among Wnt10b,β-catenin and FGF18 expressions showed that FGF18 and Wnt10b,beta catenin expression quantity were positively correlated (r=0.58,P0.05).The immunohistochemical results showed that Wnt10b andβ-catenin were expressed primarily in the epidermis,the hair placode,the hair buds,the hair nails,the inner root sheaths and the hair shafts of the mature follicles.FGF18 was not only expressed in the epidermis,the hair placode,the hair buds,the hair nails,the inner root sheaths and the hair shafts of the mature follicles,but also expressed in the dermis.So, we proposed that Wnt10b,β-catenin and FGF18 genes may be involved in regulating of SF morphogenesis through the canonical Wnt/β-catenin signaling pathways.