湖羊羔皮候选基因在不同花纹间的表达与毛囊发育特性关联的研究

Study on the Association Between the Expression of Candidate Genes in Different Waves and Hair Follicle Characteristics

【目的】研究5个候选基因在不同花纹组间的表达量与毛囊各指标的相关性,进一步了解候选基因与毛囊发育特性间的关联,筛选用于后期功能验证的基因。【方法】利用荧光定量PCR技术检测5个基因在不同组别间的表达量,结合组织学与显微观测技术,分析5个基因与毛囊发育间的关联。【结果】湖羊毛囊成群分布,湖羊大花、小花、中花多以3毛囊群居多,直径较大的为中心初级毛囊,直径相对较小的为侧部初级毛囊。湖羊大花与中花、小花初级毛囊直径呈极显著差异(P<0.01),小花与中花初级毛囊直径差异不显著(P>0.05)。中花次级毛囊直径与大花、小花次级毛囊直径呈极显著差异(P<0.01),但小花次级毛囊直径与大花次级毛囊直径差异不显著(P>0.05)。初级毛囊数大花、中花、小花间差异均不显著(P>0.05),但相同视野中大花初级毛囊数多于中花、小花。中花次级毛囊数与大花、小花次级毛囊数呈极显著差异(P<0.01),但大花、小花次级毛囊数差异不显著(P>0.05);MMP2基因在中花皮肤组织中的表达量与大花、小花分别呈极显著差异(P<0.01),在大花皮肤组织中的表达量与小花差异不显著(P>0.05);BMP7基因在大花皮肤组织中的表达量与小花呈显著差异(P<0.05),在中花皮肤组织中的表达量与大花、小花分别差异不显著(P>0.05);SFXN1基因在小花皮肤中的表达量与大花、中花分别呈显著差异(P<0.05),在大花皮肤组织中的表达量与中花差异不显著(P>0.05)。其余基因在3组个体间差异不显著;MMP2基因相对表达量与大花次级毛囊数呈显著正相关(P<0.05)。BMP7基因相对表达量与小花初级毛囊直径呈显著正相关(P<0.05),与小花次级毛囊数呈极显著正相关(P<0.01),与中花初级毛囊直径、次级毛囊数呈显著负相关(P<0.05);SFXN1基因相对表达量与大花初级毛囊呈显著负相关(P<0.05),与小花初级毛囊直径、小花次级毛囊直径分别呈显著正相关及极显著正相关,与中花初级毛囊直径呈极显著负相关。在BMP7基因表达量相同情况下,小花初级毛囊直径比中花初级毛囊直径大,小花次级毛囊数比中花次级毛囊数多,这与切片结果相符。MMP2基因在表达量相同情况下大花毛囊数多于小花,中花最少,这与切片结果一致。此外,在SFXN1基因表达量相同情况下,小花毛囊直径最大,大花次之,中花最小,这虽与切片结果不相符,但大花、小花次级毛囊数相差很小,总体来说趋势相同,结果基本相符。其余基因虽与毛囊相关指标存在关联,但在大中小花组表达差异不显著,【结论】BMP7、MMP2、SFXN1三个基因可作为湖羊早期羔皮选育的重要候选基因。

【Objective】The article aims to study the relevance between the expression of 5 candidate genes in different patterns and hair follicle characteristics, in order to learn more about the correlation between candidate genes and hair follicle characteristics and screen the genes for later functional verification. 【Method】The article detected the expression of 5 genes between different groups by RT-PCR, combined histology and microscopic observation techniques, analyzed the correlation between 5 genes and hair follicles. 【Result】 The results showed that： Hu-sheep hair follicle was distributed in groups, and it was mainly three follicles in a group, the larger diameter was the primary hair follicles, and the relatively small diameter was primary follicles.The diameter of primary follicles in large waves was significant differences with medium and small waves（P〈0.01）, and there was no significant differences between small and medium waves（P〉0.05）. The diameter of secondary follicles in medium waves was significantly differences with large and small waves（P〈0.01）, but there were no significant differences between small and large waves（P〉0.05）.The number of primary follicles was no significant differences among large, medium and small waves（P〈0.05）, but in the same vision,the number of primary follicles in large waves was more than that of medium and small waves.The number of hair follicles in medium waves was significant differences with large and small waves（P〈0.01）, but there were no significant differences between large and small waves（P〉0.05）;The expression of MMP2 among medium waves was highly significant differences with large and small waves（P〈0.01） and showed no significant differences between large and small waves（P〉0.05）. The expression of BMP7 and SFXN1 among small waves was significant differences with large and medium waves（P〈0.05） and showed no significant differences between large and medium waves; The expression of remaining genes was not significant differences among large, medium and small waves（P〉0.05）. The relative expression of MMP2 had a significant positive correlation with secondary follicle in large waves（P〈0.05）. The relative expression of BMP7 was significantly correlated with primary follicle diameter in small waves, with the number of small waves＇ secondary follicles showed a highly significant positive correlation（P〈0.01）, and negatively correlated with medium waves＇ primary follicle diameter, the number of secondary follicle（P〈0.05）; The relative expression of SFXN1 was negatively correlated with large waves＇ primary follicles（P〈0.05）, showed significantly positively related and highly significant positive correlation with small waves＇ primary and secondary follicle diameter, and in medium waves＇ primary follicles showed significant negative correlation. Under the condition of the same expression of BMP7 gene, the primary follicles＇ diameter was more than medium waves, the number of secondary follicles was more than medium waves, it was consistent with biopsy results. With the same expression of MMP2 gene, the number of large waves was more than small waves and medium was the least. It was consistent with biopsy. In addition, with the same expression of SFXN1 gene, the diameter of small waves was the largest and medium was the least. It was not match with the biopsy results, but the number of secondary follicles was slightly difference, the trend was the same, the results were basically consistent. Although other genes associated with hair follicles, their expression in large, medium, small waves were not significant.【Conclusion】 BMP7、MMP2、SFXN1 can be used as an important candidate gene for early breeding.