斑马鱼叶酸多聚谷氨酸合成酶基因的生物信息学特征及胚胎发育表达  

Bioinformatics Characteristics of Zebrafish Folylpolyglutamate Synthetase Gene and Its Expression in Embryonic Development

作　　者：黄淑莹 唐诗雨 黄婧童 李搏通 刘敏 黄四洲[2,5] 陈炳宇 Huang Shuying;Tang Shiyu;Huang Jingtong;Li Botong;Liu Min;Huang Sizhou;Chen Bingyu(School of Basic Medicine,Chengdu Medical College,Chengdu 610500,China;Development and Regeneration Key Laboratory of Sichuan Province,Chengdu Medical College,Chengdu 610500,China;School of Clinical Medicine,Chengdu Medical College,Chengdu 610500,China;School of Pharmacy,Chengdu Medical College,Chengdu 610500,China;Department of Human Anatomy,Histology and Embryology,Chengdu Medical College,Chengdu 610500,China;Department of Acupuncture and Moxibustion,Chengdu First People's Hospital,Chengdu 610095,China)

机构地区：[1]成都医学院基础医学院,成都610500 [2]成都医学院发育与再生四川省重点实验室,成都610500 [3]成都医学院临床医学院,成都610500 [4]成都医学院药学院,成都610500 [5]成都医学院人体解剖与组织胚胎学教研室,成都610500 [6]成都市第一人民医院针灸科,成都610095

出　　处：《成都医学院学报》2024年第1期1-7,23,共8页Journal of Chengdu Medical College

基　　金：国家自然科学基金(No:32070805)。

摘　　要：目的探索斑马鱼叶酸多聚谷氨酸合成酶(FPGS)的生物信息学特征及在胚胎发育中的表达模式。方法利用Ensemble数据库及ClustalW程序进行序列比对,分析斑马鱼与哺乳动物FPGS蛋白之间的氨基酸同源性;利用系统发育树分析不同物种间fpgs基因的保守性;运用反转录聚合酶链反应(RT-PCR)分析fpgs基因在斑马鱼重要器官及胚胎早期发育中的表达;运用整胚原位杂交技术检测野生型斑马鱼fpgsmRNA的表达。结果生物信息学分析显示,斑马鱼FPGS蛋白在进化上与哺乳动物同源物高度保守。RT-PCR结果显示,斑马鱼fpgs基因不仅在胚胎的各个发育阶段均有表达,且在成鱼的脑、肝和胃组织中高表达。原位杂交结果显示,fpgs基因在斑马鱼胚胎4细胞期广泛表达,而在尾芽期开始仅在卵黄合体细胞层中表达;在斑马鱼胚胎20体节期,fpgs在产生初级造血祖细胞的中间细胞团、眼睛和后脑区域呈现特异性表达;从斑马鱼胚胎受精后48~96 h,fpgs基因在其肝脏及端脑与后脑的交界处特异性表达。结论斑马鱼的FPGS蛋白与哺乳动物同源物高度保守,且fpgs基因在斑马鱼发育中的初级造血区、肝脏、后脑区及眼睛特异性表达,提示该基因在这些组织的发育中可能发挥着重要作用。Objective To investigate the bioinformatics characteristics of folylpolyglutamate synthetase(FPGS)in zebrafish and its expression patterns during embryonic development.Methods Sequence alignment was conducted by using the Ensemble database and ClustalW program to analyze the amino acid homology between zebrafish and mammalian FPGS proteins.Phylogenetic tree analysis was employed to assess the conservation of the fpgs gene among different species.Reverse transcription-polymerase chain reaction(RT-PCR)was utilized to explore the expression of zebrafish fpgs in vital organs and early embryonic development.Additionally,whole-mount in situ hybridization technique was applied to assess the expression levels of fpgs mRNA in the wild type.Results Bioinformatics analysis revealed a high conservation of zebrafish FPGS protein with its mammalian homologs in evolution.RT-PCR results demonstrated that zebrafish fpgs was not only expressed at various stages of embryonic development but also highly expressed in the brain,liver,and stomach of adult fish.In situ hybridization results revealed that fpgs exhibited maternal expression,being widely expressed during the 4-cell stage of embryonic development.However,commencing from the tailbud stage,its expression became restricted to the yolk syncytial cell layer.At the 20-somite stage,fpgs demonstrated specific expression in the intermediate cell mass generating primary hematopoietic progenitors,eyes,and the hindbrain region.Moreover,from 48 to 96 hours post-fertilization,fpgs displayed specific expression at the junction of the forebrain and hindbrain as well as in the liver.Conclusion The FPGS protein in zebrafish is highly conserved with its mammalian counterparts.The specific expression of fpgs in the primary hematopoietic region,liver,hindbrain,and eyes during zebrafish development suggests its potentially crucial role in the development of these tissues.

关 键 词：叶酸多聚谷氨酸合成酶 斑马鱼 造血 肝脏发育 

分 类 号：Q71[生物学—分子生物学]