srsf3基因在中国大鲵组织及性腺发育时期表达特征分析  

Expression Characteristics Analysis of srsf3 Gene in Tissues and Gonad During Development Stages of Chinese Giant Salamander(Andrias davidianus)

作　　者：孟方 方俊超 田海峰[2] 邵蓬 胡乔木[2] MENG Fang;FANG Jun-Chao;TIAN Hai-Feng;SHAO Peng;HU Qiao-Mu(College of Aquaculture,Tianjin Agricultural University,Tianjin 300380,China;Yangtze River Fisheries Research Institute,Chinese Academy of Fishery Sciences,Wuhan 430223,China)

机构地区：[1]天津农学院水产学院,天津300380 [2]中国水产科学研究院长江水产研究所,武汉430223

出　　处：《农业生物技术学报》2025年第4期847-856,共10页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(32373131)。

摘　　要：大鲵(Andrias davidianus)雌雄生长差异大,在性别分化过程中丝氨酸和精氨酸富集的剪接因子3(serine/arginine-rich splicing factor 3,srsf3)基因被证实参与大鲵性别分化相关基因—核受体亚家族5组A成员2(nuclear receptor subfamily 5 group A member 2,Nr5a2)选择性剪接。为探究srsf3基因在中国大鲵性别分化中的作用机制,本研究采用qPCR技术分析大鲵不同组织,不同发育时期性腺与性激素诱导后性腺中srsf3表达量的变化;通过荧光原位杂交技术(fluorescence in situ hybridization,FISH)确定srsf3基因在大鲵精巢与卵巢中的表达定位。通过前期对大鲵性腺转录组测序结果分析,srsf3基因全长为1478 bp,开放阅读框为504 bp,编码167个氨基酸。系统进化分析表明,大鲵SRSF3氨基酸序列与两栖纲热带爪蟾(Xenopus tropicalis)亲缘关系密切。qPCR结果表明,srsf3基因在大鲵卵巢中表达水平最高,垂体组织中次之,肌肉与心脏组织中表达稍低;在肝脏、脾脏、肺、胃组织中低表达。srsf3基因在大鲵性腺各时期中的表达显示,2~5年龄卵巢均显著高于精巢(P<0.05);2龄时期卵巢中表达量达最高,随后srsf3基因表达量在卵巢中呈逐年下降趋势,于4~6龄卵巢中表达量趋于稳定态势。srsf3基因表达量在性逆转雄性中最高,其次为性逆转雌性;在性逆转雌鱼卵巢中显著性高于普通雌鱼卵巢(P<0.05),性逆转雄鱼精巢中表达量显著高于普通雄鱼精巢(P<0.05)。FISH结果显示,srsf3基因在大鲵卵巢、精巢中均有表达。以上研究表明,srsf3基因参与大鲵性腺发育过程,且在大鲵性逆转过程中发挥重要作用。本研究对srsf3基因的功能解析有助于加深对两栖类性别分化机制的理解。There is a significant difference in the growth of male and female giant salamanders(Andrias davidianus),and it has been confirmed that the serine/arginine-rich splicing factor 3(srsf3)gene is involved in the selective splicing of nuclear receptor subfamily 5 group A member 2(Nr5a2),a gene related to sex differentiation in giant salamanders.To investigate the mechanism of srsf3 gene in sex differentiation of Chinese giant salamander,qPCR technology was used to analyze the changes in expression levels in different tissues,gonads at different developmental stages,and gonads induced by sex hormones;Fluorescence in situ hybridization(FISH)was used to determine the expression and localization of the srsf3gene in the testes and ovaries of giant salamanders.Based on the analysis of the transcriptome sequencing results of the giant salamander gonads,the full-length of srsf3 gene was 1478 bp,with an open reading frame of 504 bp,encoding 167 amino acids.Systematic evolutionary analysis showed that the amino acid sequence of the giant salamander SRSF3 was closely related to the Xenopus tropicalis.The qPCR results showed that the expression level of srsf3 gene was highest in the ovaries of giant salamanders,followed by pituitary tissue,slightly lower in muscle and heart tissues,and low expression in liver,spleen,lung,and stomach tissues.The expression of the srsf3 gene in the gonads of giant salamanders at different stages showed that the ovaries were significantly higher than the testes at ages 2~5(P<0.05);The expression level of srsf3 gene reached its highest in the ovaries at the age of 2,and then decreased year by year in the ovaries.The expression level tended to stabilize in the ovaries at the age of 4-6.The expression level of the srsf3 gene was highest in sex reversed males,followed by sex reversed females;The expression level of srsf3gene in the sex reversal ovaries was significantly higher than that in the ovaries(P<0.05),and the expression level in the sex reversal testes was significantly higher than that in the

关 键 词：丝氨酸和精氨酸富集的剪接因子3(srsf3)基因 中国大鲵 性逆转 表达分析 荧光原位杂交 

分 类 号：Q959.5[生物学—动物学]