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不同品系绒山羊皮肤毛囊生长发育周期中chi-miR-107-3p及其相关基因的表达分析
刘斌1,王立忠1,吴铁成1,2,马跃军1,3,赵若阳1,高玉林1,闫新刚4,李玉荣1*
0
(1.内蒙古自治区农牧业科学院, 畜牧研究所, 呼和浩特 010031;2.肉羊遗传资源评价与繁育技术内蒙古自治区工程实验室, 呼和浩特 010031;3.阿拉善白绒山羊种羊场, 内蒙古 阿拉善 750336;4.鄂尔多斯市立新实业有限公司, 内蒙古 鄂尔多斯 017000)
摘要:
为分析chi-miR-107-3p及相关基因在不同品系绒山羊皮肤毛囊生长发育周期中的表达情况,本研究分别在毛囊休止期(4月)、兴盛前期(7月)、兴盛期(9月)和退行期(1月)采集了阿拉善型绒山羊(季节性长绒)和敏盖绒山羊(常年长绒)体侧皮肤组织,通过组织切片比较分析组织形态,实时荧光定量PCR法检测chi-miR-107-3p及ADCK5、DSBC1、ARSA、FMAP4、RHBDF2ALDH3A2等6个相关基因表达量。结果表明:1)两种品系的绒山羊毛囊在形态特征上基本一致,但在同一时期生长规律不同;2)chi-miR-107-3p和RHBDF2基因在两种品系绒山羊皮肤毛囊不同发育周期的表达量呈相反趋势。在次级毛囊重建初期,chi-miR-107-3p表达量较高,而RHBDF2基因的表达量极低,随着次级毛囊重建逐步完成,chi-miR-107-3p表达量显著降低,RHBDF2基因的表达量逐渐增高。进入退行期,chi-miR-107-3p的表达量逐渐增高,而RHBDF2基因的表达量逐渐降低,休止期,chi-miR-107-3p的表达量达到最大值,此时RHBDF2基因的表达量最小,推测chi-miR-107-3p可能负向调控RHBDF2基因的表达;3)相关性分析表明,chi-miR-107-3p和RHBDF2基因表达水平均与皮肤毛囊性状显著相关(P<0.05),说明chi-miR-107-3p和RHBDF2基因均为两品系绒山羊皮肤毛囊生长发育的重要影响因子;4)ALDH3A2基因的表达量在皮肤毛囊休止期至兴盛期的过程中逐渐降低,由退行期至休止期的过程中逐渐增高。ALDH3A2基因与皮肤毛囊各性状均显著相关(P<0.05),表明高表达的ALDH3A2可能抑制两品系绒山羊毛囊的生长发育,是调控皮肤毛囊生长发育的重要影响因子,其他基因对绒山羊皮肤组织生长均无影响。本研究证实羊绒生长发育过程离不开miRNA及其相关基因参与调控,为进一步研究毛囊发育分子机理提供了参考,为实际生产中进行羊绒增产提供理论依据。
关键词:  绒山羊  组织形态  荧光定量  chi-miR-107-3p
DOI:10.11841/j.issn.1007-4333.2022.07.13
投稿时间:2021-07-09
基金项目:国家自然科学基金项目(31760653);内蒙古自治区科技计划项目(2020GG0095);内蒙古自治区科技成果转化引导项目(2020CG0100);内蒙古创新基金项目(2020CXJJM01);鄂尔多斯市绒山羊重大专项(2019);内蒙古自治区绒山羊重大专项(2017)
Expression analysis of chi-miR-107-3p and its related genes in the growth and development of hair follicle cycles in different strains of cashmere goats
LIU Bin1,WANG Lizhong1,WU Tiecheng1,2,MA Yuejun1,3,ZHAO Ruoyang1,GAO Yuling1,YAN Xingang4,LI Yurong1*
(1.Institute of Animal Husbandry, Inner Mongolia Autonomous Region Academy of Agricultural and Husbandry Science, Hohhot 010031, China;2.Engineering Laboratory for Genetic Resources evaluation and Breeding Technology of mutton sheep inInner Mongolia Autonomous Region, Hohhot 010031, China;3.Alxa white cashmere goat breeding farm, Alxa 750336, China;4.Ordos Lixin Industrial Co., Ltd., Erdos 017000, China)
Abstract:
To explore the role of chi-miR-107-3p and its related genes in the skin and hair follicle during hair follicle cycle and in different photoperiod and cashmere goats strains, the dorsal lateral skin tissues of Alashan cashmere goats(seasonal cashmere)and Mingai cashmere goats(perennial cashmere)were collected in the hair follicle rest period(April), pre-prosperity period(July), prosperity period(September)and degenerative period(January). The tissue morphology was compared and the expressions levels of chi-miR-107-3p, and mRNA of ADCK5, DSBC1, ARSA, FMAP4, RHBDF2 and ALDH3A2 were detected by real-time PCR. The results showed: 1)The morphological characteristics of hair follicles of the two cashmere goat strains were basically the same, but the growth patterns were different; 2)The expression levels of chi-miR-107-3p and RHBDF2 genes in different hair follicle cycles showed an opposite trend between two strains. In the early stage of secondary hair follicle reconstruction, the expression of chi-miR-107-3p was high, while the RHBDF2 was very low. With the completion of secondary hair follicle reconstruction, the expression of chi-miR-107-3p decreased significantly, and the expression of RHBDF2 increased gradually. Until the degenerative stage, the expression of chi-miR-107-3p started to increase gradually, while the expression of RHBDF2 gene gradually decreased. In the resting stage, the expression of chi-miR-107-3p reached to its maximum, and the expression of RHBDF2 was the lowest. It was speculated that chi-miR-107-3p may negatively regulate the expression of RHBDF2 gene; 3)Correlation analysis showed that the expression levels of chi-miR-107-3p and RHBDF2 genes were significantly correlated with skin and hair follicle traits(P<0. 05), indicating that chi-miR-107-3p and RHBDF2 genes were important factors affecting the growth and development of skin and hair follicles in cashmere goat; 4)The expression level of ALDH3A2 gene decreased gradually from telogen to anagen, and increased gradually from catagen to telogen. ALDH3A2 gene was significantly correlated with skin and hair follicle traits(P<0. 05), indicating that high expression of ALDH3A2 may inhibit the growth and development of skin hair follicles in two strains, and was important in regulating that process. In addition, other genes had no effect on the growth and development of cashmere goat skin. This study confirmed that cashmere growth and development is inseparable from the regulation of miRNA and its related genes, and provided references for further research on the molecular mechanism of hair follicle development and a theoretical basis for the actual production of cashmere production.
Key words:  Cashmere goat  tissue morphology  fluorescence quantification  chi-miR-107-3p