snx8a基因在早期斑马鱼胚胎中空间和时间特异性表达  

Spatial and temporal specific expression of snx8a genes in early zebrafish embryos

作　　者：张乙进 杨雁竹 罗红 莫大双 舒莉萍 江滟 ZHANG Yijin;YANG Yanzhu;LUO Hong;MO Dashuang;SHU Liping;JIANG Yan(Department of Clinical Microbiology and Immunology,School of Medical Laboratory Medicine,Guizhou Medical University,Guiyang 550004,Guizhou,China;Department of Immunology,School of Basic Medical Sciences,Guizhou Medical University&National and Local Joint Engineering Laboratory of Cell Engineering and Biomedical Technology&Guizhou Provincial Key Laboratory of Regenerative Medicine,Guiyang,Guizhou,550004,China;Key Laboratory of Adult Stem Cell Translation,Chinese Academy of Medical Sciences,Guiyang 550004,Guizhou,China;Laboratory Animal Resources Center,Guizhou Medical University,Guiyang 550025,Guizhou,China;Department of Microbial Immunology,Clinical Laboratory Center,the Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou,China)

机构地区：[1]贵州医科大学医学检验学院临床微生物与免疫学教研室,贵州贵阳550004 [2]贵州医科大学基础医学院免疫学教研室&细胞工程生物医药技术国家地方联合工程实验室&贵州省再生医学重点实验室,贵州贵阳550004 [3]中国医学科学院成体干细胞转化研究重点实验室,贵州贵阳550004 [4]贵州医科大学实验动物中心,贵州贵阳550025 [5]贵州医科大学附属医院临床检验中心微生物免疫科,贵州贵阳550004

出　　处：《贵州医科大学学报》2025年第1期18-23,共6页Journal of Guizhou Medical University

基　　金：国家自然科学基金资助项目(32160168,32260177);贵州省科技创新人才团队项目(CXTD[2021]002);贵州省百层次人才项目(黔科合平台人才-GCC[2023]034);贵州省科技厅基础研究计划(黔科合基础-ZK[2024]一般143)。

摘　　要：目的探索snx8a基因在斑马鱼早期胚胎中的表达情况。方法使用Snap Gene 6.0.2软件对由NCBI数据库上拷贝的人类SNX8、斑马鱼Snx8a和Snx8b氨基酸序列进行氨基酸序列比对,利用转化连接、TA克隆等分子生物学方法构建原位杂交探针质粒;通过收集不同发育时相的野生斑马鱼胚胎进行全胚胎原位杂交,观察斑马鱼早期胚胎中snx8a的空间表达情况、使用RT-qPCR检测斑马鱼早期发育不同时期snx8a基因的表达情况。结果氨基酸序列比对结果显示斑马鱼Snx8a较Snx8b与人的SNX8同源性更高;对构建的原位杂交探针质粒双酶切鉴定及测序结果显示,构建的探针质粒序列正确、质粒可用;全胚胎原位杂交结果显示,斑马鱼snx8a基因在胚胎一细胞期就开始表达,之后特异表达于斑马鱼头部及尾部造血区;RT-qPCR结果显示,snx8a基因表达逐渐升高,在胚胎12 hpf时达到峰值(P<0.05)。结论斑马鱼snx8a基因为母源性基因,其可能与斑马鱼神经发育和造血发育相关。Objective To explore the expression of Snx8a genes in early zebrafish embryos.Methods Snap Gene 6.0.2 software was used to compare the amino acid sequences of human sorting nexins 8(SNX8),zebrafish Snx8a and Snx8b copied from the NCBI database.Molecular biology methods such as transformation ligation and TA cloning were used to construct in situ hybridization(ISH)probe plasmids.The expression of snx8a in early zebrafish embryos was observed by collecting wild zebrafish embryos at different developing stages for whole embryo in situ hybridization.RT-qPCR was used to detect the expression of snx8a gene in zebrafish at different developing stages.Results The results of amino acid sequence alignment showed that the Snx8a gene had higher homology than the Snx8b in zebrafish with human SNX8.The results showed that the sequences of the constructed probe plasmids were correct and available.The results of whole embryonic ISH showed that the snx8a gene was expressed in the first cell stage of the embryo,and was specifically expressed in the hematopoietic region of the head and tail of zebrafish.RT-qPCR results showed that the expression of snx8a gene increased gradually,reaching a peak at 12 hpf in embryos(P<0.05).Conclusion The zebrafish snx8a gene is a maternally derived gene and may be related to zebrafish neural and hematopoietic development.

关 键 词：斑马鱼 snx8a基因 生物信息学分析 全胚胎原位杂交 胚胎发育 

分 类 号：R392.1[医药卫生—免疫学]