摘要
spindlin基因是减数分裂纺锤体相关因子,为了研究spindlin基因在二倍体和三倍体雌性虹鳟减数分裂过程中出现的差异,通过cDNA末端快速扩增(RACE)技术获得spindlin基因cDNA 4529 bp(GenBank登录号:MN378564),其中3′非编码区(UTR)和5′非编码区(UTR)分别长3662 bp和141 bp,开放阅读框(ORF)长726 bp,编码241个氨基酸,该蛋白质序列的相对分子量为28.3 kD,理论等电点值为5.94,无跨膜结构。同源性分析表明,虹鳟(Oncorhynchus mykiss)与银大马哈鱼(Oncorhynchus kisutch)同源最高,高达99.59%。系统发育进化树显示,虹鳟与大鳞大马哈鱼(Oncorhynchus tshawytscha)和红点鲑(Salvelinus alpinus),聚为一支。实时荧光定量(RT-PCR)结果显示,spindlin基因在二倍体雌性虹鳟卵巢、肾、肝、脾、肌、鳃、心、眼、肠和鳍组织中均有表达,其中,在卵巢中的表达量极显著高于其他组织(P<0.01)。对于二倍体雌性虹鳟,在受精后240—300d(days post fertilization,dpf)发育阶段,spindlin基因在卵巢组织中的相对表达量显著下降。对于三倍体雌性虹鳟,该基因在240—330 dpf阶段的表达量显著上升。在同一发育阶段中,spindlin基因在二倍体雌性虹鳟卵巢中的表达量较三倍体雌性虹鳟相对较高,且均存在极显著差异(P<0.01)。通过免疫组化结果发现,二倍体雌性虹鳟在240 dpf阶段,Spin蛋白在初级卵母细胞核内信号最强,在270—330 dpf阶段逐渐减弱;三倍体虹鳟卵巢在240—330 dpf发育阶段,在卵原细胞中信号逐渐增强。减数分裂异常是性腺败育的关键原因,研究结果表明三倍体虹鳟在减数分裂过程中出现异常与spindlin基因的低表达有关,这可能是卵巢发育阻滞的原因之一。
This study explored the different expression of spindlin gene,a meiosis spindle related factor,in diploid and triploid female rainbow trout meiosis.The spindlin cDNA was 4529 bp(GenBank login number:MN378564)including the 3′untranslated region(UTR)of 3662 bp and 5′untranslated region(UTR)of 141 bp.The open reading frame(ORF)was 726 bp long,encoding 241 amino acids.The relative molecular weight of the protein was 28.3 kD,the theoretical isoelectric point value was 5.94,and there was no transmembrane structure.Homology analysis showed that rainbow trout(Oncorhynchus mykiss)and silver salmon(Oncorhynchus kisutch)had the highest homology of 99.59%.Phylogenetic evolutionary trees showed rainbow trout grouped with Oncorhynchus tshawytscha and Salvelinus alpinus.RT-PCR results showed that spindlin gene was expressed in the ovary,kidney,liver,spleen,muscle,gill,heart,eye,intestine and fin tissues of diploid female rainbow trout,and the expression level in the ovary was significantly higher than that in other tissues(P<0.01).One family of female rainbow trout,spindlin gene,is particularly concerned.In the developmental stage after fertilization,240—300 days post fertilization(dpf),the relative expression of spindlin gene in ovary tissues decreased significantly.For trout triploid female rainbow,the expression level of this gene increased significantly at the 240—330 dpf stage.In the same development stage,the expression level of spindlin gene in diploid female rainbow trout was significantly higher than that of triploid(P<0.01).Immunohistochemical results showed that the signal of Spin protein in the nucleus of primary oocyte was the strongest in diploid female rainbow trout at the stage of 240 dpf,and gradually weakened at the stage of 270—330 dpf.The signal in triploid increased gradually in oocytes during the development stage of 240—330 dpf.Abnormal meiosis is the key cause of gonadal abortion.The results of this study showed that the abnormal meiosis of triploid rainbow trout was related to the low expres
作者
史秀兰
黄天晴
徐革锋
邹作宇
谷伟
程琳
刘晨斌
王炳谦
SHI Xiu-Lan;HUANG Tian-Qing;XU Ge-Feng;ZOU Zuo-Yu;GU Wei;CHENG Lin;LIU Chen-Bin;WANG Bin-Qian(Shanghai Ocean University,Shanghai 201306,China;Heilongjiang Fisheries Research Institute,Chinese Academy of Fishery Sciences,Harbin 150076,China;Harbin Academy of Agricultural Science,Harbin 150028,China)
出处
《水生生物学报》
CAS
CSCD
北大核心
2021年第1期14-21,共8页
Acta Hydrobiologica Sinica
基金
中国水产科学研究院科研业务费项目(2018HY-ZD03)
现代农业产业技术体系专项资金(CARS-46)资助。
