摘要
为探究小RNA miR-148和miR-29与民猪抗寒之间的关系,采用qPCR方法检测了miR-148和miR-29在民猪不同组织中的表达情况,同时检测PK15细胞在25℃条件下分别培养0,24,36,48,72 h后miR-148a-3p和miR-29b的表达量变化。结果表明:组织表达谱中miR-148和miR-29在卵巢和精子中的表达量最高,在其他组织中表达量最高可达胃中表达量的42倍,最低仅为胃中表达量的0.3倍,且差异极显著(P<0.01)。25℃条件下猪肾上皮细胞(PK15)随着培养时间的延长,miR-148a-3p和miR-29b的表达量变化明显,miR-148a-3p的表达量在0~36 h变化不明显,36~72 h表达量先升高后降低,miR-29b的表达量在0~72 h变化呈先降低后升高再降低再升高的波浪型变化;不同时间点靶基因DNMT1及DNMT3B的表达量变化显著,但与miR-148a-3p和miR-29b的表达量变化趋势相反。说明小RNA的表达呈现明显的组织特异性;在低温环境下,培养时间对miR-148a-3p和miR-29b及其靶基因的表达量有较为明显的影响,推测其可能与民猪的抗寒能力之间存在着一定的关系。
To explore the relationship between miR-148 and miR-29 and cold resistance of Min pig,qPCR method was used to detect the expression of miR-148 and miR-29 in different tissues of Min pig.Meanwhile,the expression of miR-148 a-3 p and miR-29 b in PK15 cells cultured at 0,24,36,48 and 72 h were detected.The results showed that the expression levels of miR-148 and miR-29 in ovary and sperm were the highest in tissue expression profile.The highest expression level in other tissues was 42 times of that in stomach,the lowest expression level was only 0.3 times of that in stomach,and the difference was extremely significant(P<0.01).At 25℃,the expression of miR-148 a-3 p and miR-29 b in PK15 cells changed significantly with the prolongation of culture time.The expression of miR-148 a-3 p did not change significantly from 0 to 36 h.The expression of miR-29 b increased first and then decreased from 36 h to 72 h.The expression of miR-29 b decreased first,then increased,then decreased and then increased at 0-72 h,showing a wavy change.There was significant difference among the detection groups.The expression of target genes DNMT1 and DNMT3 B at different time points also changed significantly,but the trend was contrary to that of miR-148 a-3 p and miR-29 b,and the difference was significant.The results showed that the expression of miRNAs showed obvious tissue specificity.Under low temperature,the change of culture time had a significant effect on the expression of miR-148 a-3 p and miR-29 b and its target genes,which may be related to cold resistance of Min pigs.
作者
王超
马红
汪亮
丁良梅
刘娣
WANG Chao;MA Hong;WANG Liang;DING Liangmei;LIU Di(College of Animal Science and Technology,Northeast Agricultural University,Harbin 150030,China;Heilongjiang Academy of Agricultural Sciences,Harbin 150086,China)
出处
《吉林农业大学学报》
CAS
CSCD
北大核心
2020年第6期679-685,共7页
Journal of Jilin Agricultural University
基金
黑龙江省自然科学基金重点项目(JJ2018ZZ0082)
国家自然科学基金项目(31872980)
