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低盐胁迫下松江鲈ATP1A3和ATP2B1基因的表达变化规律
毛非凡1, 邝杰华1, 马骞1, 周启苓1, 李昂2, 刘新富2, 庄志猛2
1.广东海洋大学水产学院 广东 湛江 524025;2.中国水产科学研究院黄海水产研究所 山东 青岛 266071
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技术检测了松江鲈的鳃、肠、肾脏和肝脏组织中2个基因在2种低盐胁迫处理(盐度渐变处理,盐度变化速率为1.1/h;盐度骤变处理,盐度变化速率为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2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。2种盐度渐变处理下,肠组织中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,盐度骤变处理下在24 h显著上升。在盐度渐变处理下,肾脏组织中2个基因的表达量均在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和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2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。
关键词:  松江鲈  盐度胁迫  ATP1A3  ATP2B1  表达变化规律
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Effects of low salinity stress on the expression profiling of ATP1A3 and ATP2B1 in the roughskin sculpin (Trachidermus fasciatus)
MAO Feifan1, KUANG Jiehua1, MA Qian1, ZHOU Qiling1, LI Ang2, LIU Xinfu2, ZHUANG Zhimeng2
1.College of Fisheries, Guangdong Ocean University, Zhanjiang, Guangdong 524025, China;2.Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao, Shandong 266071, China
Abstract:
This study explored the roles of ATPase Na+/K+ transporting subunit alpha 3 (ATP1A3) and ATPase sarcoplasmic/endoplasmic reticulum Ca2+ transporting 1 (ATP2B1) in the low-salinity stress response of Trachidermus fasciatus (roughskin sculpin). ATP1A3 and ATP2B1 gene sequences were obtained from the T. fasciatus transcriptome data, and a phylogenetic analysis was performed. Roughskin sculpins were subjected to two acute osmotic salt treatments (salinity change rates of 27/h and 1.1/h) to induce stress, and the expression patterns of the target genes in four tissues (gill, intestine, kidney, and liver) were analyzed by quantitative real-time PCR (qRT-PCR). The phylogenetic analysis showed that the ATP1A3 and ATP2B1 genes formed an independent cluster, and the T. fasciatus ATP1A3 and ATP2B1 proteins shared a high identity with those of Perciformes and Pleuronectiformes species. The teleost ATP1A3 and ATP2B1 proteins formed a single distinct lineage from other vertebrates. For both genes, each tissue had its own gene expression pattern in response to salinity. In the gills, ATP1A3 expression increased and then decreased under chronic salinity stress, but ATP1A3 expression significantly decreased under acute stress. ATP2B1 expression increased under chronic and acute stress (with a significant increase after 24 h of chronic stress). In the intestines, ATP1A3 expression significantly decreased after 24 h in both treatments, but the ATP2B1 expression significantly increased under chronic stress and increased after 24 h of acute stress. In the kidneys, the expression levels of both genes peaked after 24 h of chronic stress and increased significantly under acute stress. ATP1A3 and ATP2B1 expression was undetected in the liver under chronic stress, but under acute stress, incremental expression was detected for both genes. These results demonstrate that the ATP1A3 and ATP2B1 gene expression profiles are affected by salinity stress but not in the same way. These findings provide a foundation for future investigations into the role of ATP1A3 and ATP2B1 in fish osmotic pressure regulation and the molecular mechanisms underlying migratory fish salinity adaptation.
Key words:  Trachidermus fasciatus  Salinity stress  ATP1A3  ATP2B1  Gene expression patterns