ERR-dsRNA对罗氏沼虾卵巢中ERR及生殖相关基因表达的影响

摘要

[Objective] To investigate the functions of Estrogen-related receptor (ERR) in the reproductive process in female giant freshwater prawn (Macrobrachium rosenbergii).[Method] The expressions of ERR were knock-down by injecting double-stranded RNA (dsRNA). Ovary transcriptome sequencing was curried out from the ERR-dsRNA injected and un-injected control groups to screen differentially expressed genes related to reproduction and their expression pattern during ovarian development were detected by qPCR.[Result] The levels of ERR mRNA and ERR protein were significantly decreased after injecting ERR-dsRNA. Among the differentially expressed Unigenes obtained from ERR-dsRNA injected and control ovary transcriptomes, 9 Unigenes were enriched from reproduction GO term, 3 Unigenes were enriched to reproductive process GO term and 21 were identified to have been enriched to oocyte meiosis Pathway term. There are 9 Unigenes with large fold-changes in differential expression related to reproduction. Of these, 5 Unigenes were verified to be differential expressions between ERR-dsRNA interference and control groups by qPCR. Among the 5 differentially expressed Unigenes, 3 Unigenes were differentially expressed during ovarian development process. 2 of these Unigenes (CL7112.Contig4_All and Unigene4600_All) were associated with the oocyte meiosis and 1 unigene (CL4888.Contig2_All) was related to the synthesis and release of reproduction-associated hormones.[Conclusion] The function of ERR is likely to regulate ovarian development by regulating the oocyte meiosis as well as the synthesis and release of reproduction-associated hormones in M.rosenbergii.%[目的]研究雌激素相关受体(Estrogen-related receptor,ERR)在雌性罗氏沼虾生殖过程中的功能.[方法]通过注射双链RNA(Double-stranded RNA,dsRNA)干扰罗氏沼虾(Macrobrachium rosenbergii)ERR的表达,对干扰组和对照组的卵巢样本进行转录组测序,筛选与生殖相关的差异表达基因,并通过qPCR检测它们在卵巢发育过程中的表达.[结果]注射ERR-dsRNA后,卵巢中ERR mRNA和ERR蛋白的表达显著下降.ERR-dsRNA干扰组和对照组卵巢转录组测序获得的差异表达Unigene中,9条被富集到生殖GO条目,3条被富集到生殖过程GO条目,21条被富集到卵母细胞减数分裂Pathway条目.这些生殖相关的Unigene中差异倍数较大的有9条,经qPCR验证,5条在ERR干扰组和对照组间的表达差异有统计学意义.这5条Unigene中有3条在卵巢发育过程中的表达差异有统计学意义,其中2条(CL7112.Contig4_All和Unigene4600_All)与卵母细胞减数分裂有关,1条(CL4888.Contig2_All)与生殖相关激素的合成及释放有关.[结论]ERR可通过调控卵母细胞减数分裂和生殖相关激素的合成与释放调控罗氏沼虾卵巢发育.