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引用本文:李旋,冯艳微,袁波,姜绪,何金霞,刘相全.皱纹盘鲍(Haliotis discus hannai)WNT4基因生物信息学分析及与性腺发育相关性的研究.海洋与湖沼,2021,52(4):1019-1028.
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皱纹盘鲍(Haliotis discus hannai)WNT4基因生物信息学分析及与性腺发育相关性的研究
李旋1,2,3, 冯艳微4, 袁波1,2,3, 姜绪3, 何金霞3, 刘相全3
1.上海海洋大学 水产科学国家级实验教学示范中心 上海 201306;2.上海海洋大学 上海水产养殖工程技术 研究中心 上海 201306;3.山东省海洋资源与环境研究院 山东省海洋生态修复重点实验室 烟台 264006;4.鲁东大学农学院 烟台 264025
摘要:
WNT4作为WNT基因家族的重要成员,在动物的性腺发育过程中起重要调控作用。从皱纹盘鲍(Haliotis discus hannai)转录组文库中获得WNT4基因的cDNA序列,该序列长2002 bp,包括5'非编码区(5'UTR)342 bp,3'非编码区(3'UTR)595 bp,开放阅读框(ORF)长度为1 071 bp,可以编码356个氨基酸。该蛋白序列与福寿螺(Pomacea canaliculata)、长牡蛎(Crassostrea gigas)、栉孔扇贝(Azumapecten farreri)和厚壳贻贝(Mytilus coruscus)的同源性分别为71.86%、70.92%、67.51%和65.32%。通过qRT-PCR分析WNT4基因在雌雄皱纹盘鲍不同月份不同组织中的表达特点,其结果显示,除肝胰腺外,WNT4基因在其雌雄个体不同月份的鳃、外套膜、肌肉和性腺中均有表达;在雌性个体性腺中,WNT4基因的表达量以7月份为最高,显著高于其他月份;其次为5、8月,显著高于6、9月的表达量;5月与8月、6月与9月之间均差异不显著。在雄性个体性腺中,不同月份WNT4的表达量7月>8月>9月>5月>6月;7、8月的表达量均显著高于其他月份,9月显著高于6月的表达量,与5月差异不显著,5月表达量与6月差异不显著。此外,WNT4基因在精巢的表达量均高于卵巢。原位杂交(ISH)结果显示,WNT4在5—9月雌性皱纹盘鲍性腺的卵母细胞和雄性性腺的结缔组织均有明显的蓝紫色杂交信号。皱纹盘鲍WNT4基因在各个组织均有表达,表明其的表达特点是广泛性的,参与多种组织细胞的生命过程;在不同月份性腺中的表达特征,推测其可能在两性性腺发育过程中发挥作用。
关键词:  皱纹盘鲍  WNT4基因  性腺发育  表达分析  原位杂交
DOI:10.11693/hyhz20201200349
分类号:Q78
基金项目:山东省农业良种工程项目,2017LZGC009号。
附件
BIOINFORMATICS ANALYSIS OF WNT4 GENE OF HALIOTIS DISCUS HANNAI AND ITS CORRELATION TO GONADAL DEVELOPMENT
LI Xuan1,2,3, FENG Yan-Wei4, YUAN Bo1,2,3, JIANG Xu3, HE Jin-Xia3, LIU Xiang-Quan3
1.National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai 201306, China;2.Shanghai Engineering Research Center of Aquaculture, Shanghai Ocean University, Shanghai 201306, China;3.Shandong Provincial Key Laboratory of Restoration for Marine Ecology, Shandong Marine Resource and Environment Research Institute, Yantai 264006, China;4.Ludong University, College of Agriculture, Yantai 264025, China
Abstract:
As an important member of the WNT gene family, WNT4 plays a pivotal role in development processes of many animals. In the present study, the cDNA sequence of WNT4 gene was obtained from the transcriptome library of Haliotis discus hannai. The length of the WNT4 gene was 2 002 bp, including 5'-UTR 342 bp and 3'-UTR 595 bp. The open reading frame (ORF) was 1 071 bp, encoding a polypeptide of 356 amino acids. As revealed in the real-time quantitative PCR analysis in different tissues of male and female abalone in different months, WNT4 gene was expressed in gonad, muscle, mantle and gill, but hepatopancreas. The expression level of WNT4 gene in gonad of female was the highest in July (P<0.05), followed by May and August, and it was significantly higher than that in June and September. There were no significant differences between May and August, and June and September. In gonads of male individuals, the expression of WNT4 in different months was in a descending order of July > August > September > May > June. The expression level in July and August were significantly higher than that in other months, and that in September was significantly higher than that in June, but not significantly different from that in May. In addition, the expression level of WNT4 gene in testis was higher than that in ovaries. In situ hybridization results showed that WNT4 had obvious blue-purple hybridization signals in male and female gonads in different months. The expression characteristics of WNT4 in many tissues of H. discus hannai indicate that it has a wide range of tissue expression characteristics and participates in the life process of many tissue cells in the form of signal molecules. From the gonad expression characteristics, it is speculated that it may play a role in the development of gonadal development.
Key words:  Haliotis discus hannai  WNT4 gene  gonadal development  expression analysis  in situ hybridization
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