SHH在美利奴羊与小尾寒羊背部皮肤表达差异的研究

Differences in SHH expression in the back skin of Merino sheep and Small-tail Han sheep

为了研究音猬因子(sonic hedgehog, SHH)在不同品种绵羊背部皮肤中的表达差异,以探索SHH与羊毛弯曲形成的关系,选取美利奴羊和小尾寒羊作为研究对象,利用HE染色法观察2种绵羊背部皮肤的组织结构,采用免疫组织化学技术、实时荧光定量PCR和Western blot方法探究2种绵羊背部皮肤中SHH蛋白和基因水平的相对表达量差异。结果显示:HE染色显示美利奴羊与小尾寒羊背部皮肤毛囊的组织结构存在毛囊数量及弯曲程度等差异;免疫组织化学试验显示,SHH蛋白在美利奴羊和小尾寒羊背部皮肤中均有表达,在美利奴羊背部皮肤中的毛乳头、毛基质、内外根鞘、皮脂腺和表皮细胞中显著表达,在小尾寒羊背部皮肤中的毛基质、内外根鞘和皮脂腺中显著表达;光密度分析显示,美利奴羊背部皮肤组织中SHH蛋白的表达量显著高于小尾寒羊(P<0.001);实时荧光定量PCR显示,美利奴羊SHH mRNA相对表达水平显著高于小尾寒羊(P<0.01);Western blot显示,美利奴羊SHH蛋白相对表达水平显著高于小尾寒羊(P<0.01)。提示:羊毛弯曲的形成与其组织结构有关,并受SHH的调控。

In order to determine the differences in the expression of Sonic hedgehog(SHH) in the back skin of different breeds of sheep, and to explore the relationship between SHH and wool bending, two different breeds of sheep, Merino sheep and Small-tail Han sheep were used as the research objects. Hematoxylin-eosin staining(HE) was used to observe the tissue structure of the back skins of the two sheep breeds, and immunohistochemistry, real-time PCR and Western blot were used to explore the relative expression levels of SHH protein and its gene levels in the back skins of the sheep. The HE results showed that there were differences in the number and degree of curvature of the hair follicles in the back skins of Merino sheep and Small Tail Han sheep. The results of immunohistochemistry showed that the SHH protein did express in the back skins of the sheep breeds;the protein significantly expressed in the dermal papilla, hair matrix, inner and outer root sheaths, sebaceous glands and epidermal cells in the back skin of Merino sheep, but it significantly expressed only in the hair matrix, inner and outer root sheaths and sebaceous glands in the back skin of Small-tail Han sheep. The results of the optical density analysis showed that the expression of the SHH protein in the back skin tissue of Merino sheep was significantly higher than in that of Small-hail Han sheep(P<0.001). Real-time PCR results showed that the relative expression level of the SHH mRNA in Merino sheep was significantly higher than that in Small-hail Han sheep(P<0.01), and the results of Western blot showed that the relative expression of the SHH protein in Merino sheep was significantly higher than that in Small Tail Han sheep(P<0.01). The above results suggested that the formation of wool bending was related to its organizational structure and was regulated by SHH.