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梅花鹿茸生长过程中顶端不同组织COL1A1基因启动子DNA甲基化模式及差异分析
张芙蕊,韩若冰,郭梦雅,赵洵武,李和平
0
(东北林业大学 野生动物与自然保护地学院, 哈尔滨 150040)
摘要:
为探究COL1A1基因表达的Ⅰ型胶原蛋白α1链在组成生物体结构和骨发育中的重要作用,以梅花鹿茸生长过程的小鞍子(前期)、二杠(中期)和三杈茸(后期)3个典型时期鹿茸顶端组织及其茸皮、间充质、前软骨和软骨4个组织层为试验材料,采用亚硫酸氢盐测序法(BSP技术),从时空角度研究鹿茸生长过程中顶端不同组织COL1A1基因启动子区DNA甲基化模式及其相互间DNA甲基化差异。结果显示:1)COL1A1基因在前、中、后期的茸皮组织中的甲基化率分别为(9.73±0.92)%、(7.60±0.69)%和(3.73±0.23)%;间充质组织中的甲基化率分别为(3.20±0.40)%、(1.33±0.23)%和(1.60±0.69)%;前软骨组织中的甲基化率分别为(4.67±0.83)%、(2.53±0.46)%和(2.67±0.23)%;软骨组织中的甲基化率分别为(5.60±0.40)%、(2.80±0.40)%和(2.27±0.61)%。2)COL1A1基因启动子区的甲基化区域共有25个CG位点,在17个CG位点上均发生了不同程度的甲基化。3)对COL1A1基因启动子区DNA甲基化差异分析发现,相同时期中,茸皮组织甲基化率最高,间充质组织甲基化率最低;相同组织中,中期比前期显著下调;CG位点的比较中,前期与中期的CG位点差异程度较大。综上,COL1A1基因在快速生长期,以及在间充质组织、前软骨组织、软骨组织中对鹿茸生长具有促进作用。在DNA甲基化水平上探究梅花鹿鹿茸不同组织的时空表观遗传差异,为鹿茸快速生长与骨化机制的研究,以及哺乳动物组织再生和器官修复等领域的研究提供了科学参考。
关键词:  梅花鹿(Cervus nippon)  鹿茸  COL1A1基因  DNA甲基化  BSP技术
DOI:10.11841/j.issn.1007-4333.2022.10.14
投稿时间:2022-01-13
基金项目:国家重点研发计划项目(2018YFC1706601)
DNA methylation and differential analysis of COL1A1 gene promoter in the different tissues of antler top during the antler growing in sika deer(Cervus nippon)
ZHANG Furui,HAN Ruobing,GUO Mengya,ZHAO Xunwu,LI Heping
(College of Wildlife and Protected Area, Northeast Forestry University, Harbin 150040, China)
Abstract:
To explore the importance of type I collagen α1 chain expressed by COL1A1 gene in the organization of organism structure and bone development, four antler top layered tissues, e. g. dermis tissue, mesenchyme tissue, precartilage tissue and cartilage tissue, in the three processes of sika deer antler growth, e. g. small saddle(earlier stage), two poles(middle stage), and three bifurcation(later stage), were used as experimental materials. The bisulfite sequencing(BSP technology)was used to study the DNA methylation patterns of the COL1A1 gene promoter region and analyze the differences in DNA methylation among different antler top tissues from the perspective of time and space. The results showed that: 1)The methylation rates of COL1A1 gene in the dermis tissues of the earlier stage, middle stage and later stage were(9. 73±0. 92)%, (7. 60±0. 69)% and(3. 73±0. 23)%; the methylation rates in the mesenchyme tissues were(3. 20±0. 40)%, (1. 33±0. 23)% and(1. 60±0. 69)%; the methylation rates in the precartilage tissues were(4. 67±0. 83)%, (2. 53±0. 46)% and(2. 67±0. 23)%; the methylation rates in the cartilage tissues were(5. 60±0. 40)%, (2. 80±0. 40)% and(2. 27±0. 61)%, respectively. 2)There were 25 CG sites in the promoter region of the COL1A1 gene, and different degrees of methylation occurred at these 17 CG sites. 3)The DNA methylation difference analysis of COL1A1 gene promotor region showed that the methylation rate of antler skin was the highest and that of mesenchymal tissue was the lowest in the same period; In the same tissue, it was significantly higher in the middle than in the earlier stage; In the comparison of CG loci, there was a great difference between the earlier and middle CG loci In conclusion, COL1A1 gene can promote the growth of pilose antler in the rapid growth period, as well as in mesenchymal tissue, anterior cartilage tissue and cartilage tissue. This study explored the epigenetic difference of time and space in different tissues of sika deer antler at the level of DNA methylation, which provided a scientific reference for the study of the rapid growth and ossification mechanism of the antler as well as the research of mammalian tissue regeneration and organ repair.
Key words:  Sika deer(Cervus nippon)  Antler  COL1A1 gene  DNA methylation  BSP technology