湖羊BMP7基因遗传多态、表达及与羔皮毛囊性状的关联

doi:10.3864/j.issn.0578-1752.2014.09.016

摘要/Abstract

摘要： 【目的】研究BMP7基因对湖羊不同花纹形成的影响及其与毛囊性状的发育可能存在的调控机制。【方法】利用PCR-SSCP方法在205个样本中检测BMP7基因外显子1、2、3的单核苷酸多态性；通过组织学方法和显微观察法对湖羊皮肤毛囊进行组织学观察，对不同花纹皮肤组织中初级毛囊直径、次级毛囊直径、初级毛囊数、次级毛囊数进行统计分析；利用荧光定量PCR技术检测BMP7基因在大花、中花、小花间的相对表达量，用SPSS17.0软件进行差异显著性分析，探讨BMP7基因对湖羊不同花纹形成及其毛囊性状可能存在的调控机制。【结果】PCR-SSCP方法检测BMP7基因外显子1、2、3的单核苷酸多态性，结果均未发现多态性；在组织切片分析中，湖羊大花初级、次级毛囊直径均大于中花、小花，小花初级、次级毛囊直径均介于大花、中花之间；大花次级毛囊数多于中花、小花，而小花次级毛囊数介于大花、中花之间。通过差异显著性分析可知，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）；qRT-PCR结果显示，在同一时期BMP7基因在大花、中花中的表达量高于小花，且在大花、小花毛囊组织中表达量差异显著（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和中花次级毛囊数呈显著负相关（P＜0.05），这与组织学与显微镜观察的结果相符。【结论】未发现BMP7基因存在单核苷酸多态性，但BMP7基因表达量与毛囊部分指标存在相关性且与组织学观察结果相一致，可初步推测BMP7基因可能参与毛囊发育，调控被毛生长。

关键词: 湖羊 , BMP7 , 多态性 , 毛囊

Abstract: 【Objective】 The article aims to study the influence of BMP7 on the formation of Hu sheep’s different patterns and the possible regulatory mechanisms on the development of hair follicles. 【Method】 BMP7 gene in a sample of 205 single nucleotide polymorphisms in exons 1,2 and 3 were detected by PCR-SSCP. Hu sheep’s hair follicles were observed by histology and microscopic examination, statistical analysis on the diameter of primary and secondary hair follicles, the number of primary and secondary hair follicles were statistically analyzed. The relative expression of BMP7 among large, medium and small waves was detected by fluorescence quantitative PCR, and SPSS 17.0 software was used to conduct the significant difference analysis, to investigate the BMP7 gene’s regulatory mechanisms of the formation of Hu sheep’s different patterns and hair follicles traits. 【Result】 The results showed that by using PCR-SSCP method for detecting the single nucleotide polymorphisms of BMP7 gene’s 3 exons and no polymorphism was found. In histological analysis, the diameter of large waves primary hair follicles was more than medium and small waves, the diameter of small waves primary and secondary hair follicles was between large and medium waves. The number of large waves secondary hair follicles was more than medium and small waves, while the small waves were between large and medium waves. Through significant difference analysis, the difference in diameter of primary follicles in large waves was highly significant with medium and small waves (P＜0.01), the difference in diameter of primary follicle between large and small waves was not significant (P＞0.05). The difference in diameter of secondary follicles in medium waves was highly significant with large and small waves (P＜0.01), but the difference in diameter of secondary follicle between large and small waves was not significant (P＞0.05). The difference in number of primary follicles among large, medium, and small waves was not significant (P＞0.05), but in the same vision, the number of primary follicles among large waves was higher than medium and small waves. The difference in number of secondary follicles in medium waves was highly significant with large and small waves (P＜0.01), but the difference in number of secondary follicles between large and small waves was not significant (P＞0.05). qRT-PCR results showed that in the same period, the expression of BMP7 in small waves was higher than large waves, and showed a significant difference between large and small waves (P＜0.05). The relative expression of BMP7 was significantly correlated with the primary follicle diameter in small waves, and showed a highly significant positive correlation (P＜0.01)with the number of small waves’ secondary follicles, and negatively correlated with middle waves’ primary follicle diameter, the number of secondary follicle (P＜0.05). The expression was consistent with biopsy results. 【Conclusion】 Single nucleotide polymorphism was not found in BMP7 gene, however, the expression of BMP7 was correlated with hair follicles indexes and consisted with the histological observation. It is speculated that BMP7 gene may be involved in the development of hair follicle and regulation of hair growth.

Key words: Hu sheep , BMP7 , polymorphism , hair follicle

殷金凤1, 倪蓉1, 王庆增1, 孙伟1, 丁家桐1, 张有法2, 陈玲2, 吴文忠2, 周洪3. 湖羊BMP7基因遗传多态、表达及与羔皮毛囊性状的关联[J]. 中国农业科学, 2014, 47(9): 1811-1818.

YIN Jin-Feng-1, NI Rong-1, WANG Qing-Zeng-1, SUN Wei-1, DING Jia-Tong-1, ZHANG You-Fa-2, CHEN Ling-2, WU Wen-Zhong-2, ZHOU Hong-3. The Genetic Polymorphism, Expression of BMP7 Gene and Its Relationship with Lamb Skin Follicle Traits in Hu Sheep[J]. Scientia Agricultura Sinica, 2014, 47(9): 1811-1818.

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https://www.chinaagrisci.com/CN/Y2014/V47/I9/1811

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