文章摘要
低盐胁迫下松江鲈ATP1A3和ATP2B1基因的表达变化规律
Effects of low salinity stress on the expression profiling of ATP1A3 and ATP2B1 in the roughskin sculpin (Trachidermus fasciatus)
投稿时间:2020-07-26  修订日期:2020-08-19
DOI:
中文关键词: 松江鲈  盐度胁迫  ATP1A3  ATP2B1  表达变化规律
英文关键词: Trachidermus fasciatus  salinity stress  ATP1A3  ATP2B1  gene expression patterns
基金项目:国家自然科学基金项目(31772828)和青岛市应用基础研究计划项目(14-2-4-15-jch)
作者单位E-mail
毛非凡 广东海洋大学 435230060@qq.com 
马骞 广东海洋大学 maq@gdou.edu.cn 
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中文摘要:
      为了探究Na+/K+-ATP酶和Ca2+-ATP酶在松江鲈(Trachidermus fasciatus)应对低盐胁迫过程中的调节作用,本研究基于前期转录组数据,获取目标基因ATP1A3(Na+/K+-ATP酶α3亚基基因)和ATP2B1 (Ca2+-ATP酶1基因)的序列信息并进行了系统进化分析。利用实时荧光定量PCR技术检测了松江鲈鳃、肠、肾脏和肝脏组织中2个基因在两种低盐胁迫处理(盐度渐变处理,盐度变化速率1.1/h;盐度骤变处理,盐度变化速率27/h。)下不同时间点(0 h、12 h、24 h和48 h)的表达水平。系统进化分析结果表明,ATP1A3和ATP2B1基因分别聚类形成独立分支;在各基因分支中,松江鲈与已报道的鲈形目和鲽形目等鱼类共同聚在硬骨鱼类分支中。在两种低盐胁迫处理下,2个基因在鳃、肠、肾脏和肝脏组织中的表达量呈现不同的变化趋势。鳃组织中ATP1A3表达量在盐度渐变处理下先上升后下降,ATP2B1表达量仅在24 h显著升高;盐度骤变处理下ATP1A3表达量显著下降,ATP2B1表达量显著上升。肠组织中两种盐度渐变处理下ATP1A3表达量均在24 h显著下降;ATP2B1表达量在盐度渐变处理下显著上升,盐度骤变处理下24 h显著上升。肾脏组织中2个基因的表达量在盐度渐变处理下均在24 h显著上升至最大值;ATP1A3表达量在盐度骤变处理下显著上升,ATP2B1表达量在12 h、48 h显著上升。肝脏组织中2个基因的表达量在盐度渐变处理下均无显著变化;盐度骤变处理下ATP1A3表达量持续显著上升,ATP2B1表达量在48 h显著上升。结果表明低盐胁迫处理显著影响了ATP1A3和ATP2B1基因的表达水平,但两个基因的表达量变化规律存在显著性差异。上述结果为探讨Na+/K+-ATP酶和Ca2+-ATP酶在鱼类渗透压调节过程中的作用及洄游性鱼类适应盐度变化的分子调控机制提供了理论依据。
英文摘要:
      The aim of this study was to explore the role of ATP1A3 (ATPase Na+/K+ transporting subunit alpha 3)and ATP2B1 (ATPase sarcoplasmic/endoplasmic reticulum Ca2+ transporting 1)in stress-responsive regulation of roughskin sculpin (Trachidermus fasciatus) under low salinity stress. Firstly, sequences of these genes were obtained from transcriptome data of T. fasciatus, and then phylogenetic analysis was performed. In this study, fish were respectively subjected to two different acute osmotic treatments (salinity changing rate of 27 ppt/h and 1.1 ppt/h), expression patterns of the two target genes in the four target tissues (gill, intestine, kidney and liver) were examined using qRT-PCR. As a result, phylogenetic analysis revealed that ATP1A3 and ATP2B1 genes respectively formed an independent cluster, in which T. fasciatus ATP1A3 and ATP2B1 protein shared high identity with those of Perciformes and Pleuronectiformes species, teleost ATP1A3 and ATP2B1 proteins formed a single lineage distinct from those of other vertebrates. Tissue-specific gene expression patterns of all the two target genes showed that each tissue possesses its own gene expression pattern in response to salinity changes. In the gill, expression of ATP1A3 was increased first then decreased under the relative chronic salinity stress,the expression of ATP2B1 increased significantly at 24 h; under the acute salinity stress, the expression of ATP1A3 was significantly decreased and the expression of ATP2B1 was significantly increased. In the intestine, expression of ATP1A3 was significantly decreased at 24 h under both treatments,but the expression of ATP2B1 was increased significantly under the relative chronic salinity stress and at 24 h in response to the acute chronic salinity stress. In the kindey, expression of both genes reached the highest level at 24 h under the relative chronic salinity stress; under the acute chronic salinity stress expression of both genes were increased significantly.Expression of neither ATP1A3 nor ATP2B1 was detected under the relative chronic salinity stress in the liver; under the acute salinity stress, an increment of gene expression was detected in both the two target genes. These results demonstrate that significant affected ATP1A3 and ATP2B1 expression profiling in the salinity stress treatments, but there are significant differences in the gene expression patterns of both genes. These findings could provide a theoretical foundation for revealing the role of Na+/K+-ATPase and Ca2+-ATPase in fish osmotic pressure regulation and the molecular mechanism of migratory fish salinity adaptation.
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