三角帆蚌17β-HSD11基因的表达特征及17β-雌二醇对其表达的影响

Expression analysis of 17β-HSD11 gene in Hyriopsis cumingii and effect of 17β-estradiol on its expression

摘要:

通过RACE技术、实时荧光定量(qRT-PCR)技术、原位杂交和17β-雌二醇注射来探究17β-HSD11基因在三角帆蚌性腺发育和性激素合成中的表达特性和作用。结果显示:17β-HSD11基因的cDNA全长为1 134 bp, 其中,5' UTR 40 bp,开放阅读框(ORF)923 bp,3' UTR 171 bp,编码307个氨基酸。17β-HSD11基因在肝胰腺、性腺中表达量较高,且卵巢极显著高于精巢。原位杂交结果显示雌性三角帆蚌卵母细胞、滤泡膜和卵膜上均存在杂交信号。注射不同质量浓度17β-雌二醇之后发现17β-HSD11基因在雌雄性腺中的相对表达量均被抑制,其中:低质量浓度注射后,17β-HSD11基因的表达量在卵巢中下降33.38%,在精巢中下降37.74%;而高质量浓度注射后,17β-HSD11基因的表达量在卵巢中下降57.78%,在精巢中下降61.31%。由此推测17β-HSD11可能与三角帆蚌性腺发育和雌激素合成相关。

Abstract:

The effects of 17β-HSD11 gene on gonad development and sex hormone synthesis in H. cumingii were investigated by RACE (Rapid-amplificiantion of cDNA ends), real-time quantitative (qRT-PCR), in situ hybridization (ISH) and 17β-estradiol injection. The results showed that the full length of 17β-HSD11 cDNA was 1 134 bp, including 5' UTR 40 bp, open reading frame (ORF) 923 bp, 3' UTR 171 bp. It coded 307 amino acids. 17β-HSD11 was highly expressed in the hepatopancreas and the gonad, and the ovary was significantly higher than the testis. ISH results showed that there were hybridization signals on oocyte, follicular membrane and egg membrane. The expression of 17β-HSD11 gene was inhibited in both male and female gonads after injection of 17β-estradiol at different mass concentrations. The expression of 17β-HSD11 gene was decreased by 33.38% in ovary and 37.74% in testis after injection of 17β-estradiol at low mass concentrations. Moreover, after high mass concentration injection, the expression of 17β-HSD11 gene was decreased by 57.78% in ovary and 61.31% in testis. From the above results, it was speculated that 17β-HSD11 was related to gonadal development and estrogen synthesis in H. cumingii.