摘要
为探究钙调控因子在硬骨鱼类钙代谢过程中发挥的作用,本研究选取细胞内Ca^(2+)信号传导的关键调控因子-钙调素(Calmodulin,CaM)为研究对象,通过对虹鳟(Oncorhynchus mykiss)幼鱼腹腔注射氯化卡咪唑铵(Calmidazolium chloride,CMZ)抑制CaM活性,并在注射后6 h采集实验组及对照组(腹腔注射同体积生理盐水)虹鳟鳞片组织,利用转录组测序技术分析实验组mRNA的表达变化。结果显示:转录组测序共获得376.31 M Raw reads,Q30碱基百分比均在93.26%以上;实验组鳞片组织中共筛选出3730个差异表达基因(DEGs),其中1227个基因表达上调,2503个基因表达下调。GO注释结果显示,上述DEGs主要富集到细胞过程、生物调控、细胞器、结合和生物过程调控等功能;KEGG通路富集分析结果显示,DEGs显著富集的前3条通路依次为核糖体通路、Rap1信号通路和轴突导向通路,此外,DEGs还显著富集于破骨细胞分化、TGF-beta等与骨骼发育、钙代谢等密切相关的信号通路。本研究所获得的差异表达基因信息及其功能注释和通路富集结果可为阐明鱼类钙代谢的调控机理提供新素材。
Calcium metabolism is mainly achieved through the complex interaction between various calcium regulatory factors.Calmodulin(CaM)is a crucial regulator of intracellular Ca^(2+)signal transduction.Juveniles of Oncorhynchus mykiss were intraperitoneally injected with the CaM antagonist-Calmidazolium chloride(CMZ)to inhibit CaM activity and to explore the role of CaM in the process of calcium metabolism in teleosts.Six hours after the injection,the scale tissues of O.mykiss in the experimental group and the control group(intraperitoneal injection of the same volume of normal saline)were collected,and the mRNA levels in the scales were analyzed by transcriptome sequencing.The sequencing was performed using the Illumina Novaseq 6000 sequencing platform.The results showed that a total of 376.31 M raw reads were obtained,and the percentage of Q30 bases was above 93.26%.By setting the screening conditions for the significant differentially expressed genes(DEGs)as log2|fold change|≥1 and P<0.05,a total of 3730 DEGs were identified in the scale tissues of the experimental group,where 1227 were up-regulated and 2503 were down-regulated.Gene Ontology(GO)function annotation analysis showed that the aforementioned DEGs were mainly annotated in biological functions such as cellular process,biological regulation,organelle,binding,and regulation of biological process.Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis showed that the top three enriched pathways were Ribosome,Rap1 signaling pathway,and Axon guidance.Furthermore,DEGs were also significantly enriched in osteoclast differentiation,TGF-beta,and other signaling pathways closely related to bone development and calcium metabolism.The quantitative real-time PCR(qRT-PCR)was used to examine the expression of six randomly selected DEGs,and the results of the qRT-PCR and transcriptome data were consistent,indicating the reliability of the sequencing.The information on the DEGs and their functional annotation and pathway enrichment data from this study could provide new materials for exploring the regulation mechanism of calcium in teleost fish.
作者
任亦柯
王刘永
周启苓
吴雨薇
马骞
陈刚
REN Yike;WANG Liuyong;ZHOU Qiling;WU Yuwei;MA Qian;CHEN Gang(College of Fisheries,Guangdong Ocean University,Zhanjiang 524088,Guangdong,China)
出处
《淡水渔业》
北大核心
2025年第1期3-10,共8页
Freshwater Fisheries
基金
国家自然科学基金项目(31772828)
广东海洋大学科研启动经费资助项目(R19022)。
