四种氨基酸神经递质对斑马鱼胚胎发育和成体组织再生的双重影响  

Dual effects of four amino acid neurotransmitters on Danio rerio embryonic development and adult tissue regeneration

作　　者：李心雨 刘亚平 杜新路 严继舟[1] LI Xin-yu LIU Ya-ping DU Xin-lu YAN Ji-zhou(Department of Biology, College of Fisheries and Life Sciences, Shanghai Ocean University, Shanghai 201306, China)

机构地区：[1]上海海洋大学水产与生命学院生物系,上海201306

出　　处：《淡水渔业》2017年第5期58-63,101,共7页Freshwater Fisheries

基　　金：上海市教委大学生科研创新项目(B-5106-13-0001);上海市教委水产高峰学科项目(A1-2035-17-0001B3-12)

摘　　要：筛选小分子化合物促进组织再生仍然是再生医学的一个巨大挑战。前期研究提示神经递质在斑马鱼(Danio rerio)下颌再生过程中发挥着重要作用。本实验分别剪切Flk-GFP转基因斑马鱼成体下颌组织和1月龄幼鱼尾鳍,然后将其置入四种神经递质(牛磺酸、谷氨酸、甘氨酸以及γ-氨基丁酸)的浓度梯度环境中,培养5d后取成鱼下颌制作冰冻切片进行观察检测新生血管荧光量,对幼鱼尾鳍再生进行长度测量。结果显示甘氨酸和γ-氨基丁酸对损伤组织的再生表现出促进作用,而牛磺酸和谷氨酸表现出在低浓度时促进与高浓度的抑制作用。胚胎培养和早期胚胎细胞增生的MTT试验进一步验证了氨基酸神经递质在不同浓度所表现出的细胞营养和毒性作用。结果提示有必要建立一个多层面包括从细胞-胚胎-幼鱼和成鱼的斑马鱼检测系统,能有效筛选再生相关小分子化合物及其合适浓度。Screening small molecules targeting tissue regeneration remains challenge in regenerative medicine. The previous investigations have indicated that neurotransmitters play important roles in the process of regeneration in Danio rerio jaw. Flk - GFP transgenic D. rerio adult jaw and 1 - month - old caudal fins were separately cut in this study, and then rear the damaged fishes for 5 days in the presence of four amino acid neurotransmitters （ taurine, glutamate, glycine, and γ- aminobutyric acid （GABA） ） at gradient concentrations. Their effects on tissue regeneration were compared by measuring GFP intensity of cryosections, or by measuring the caudal fin regeneration length. The results showed that glycine and GABA promote damaged tissue regeneration, while taurine and glutamate exhibit promotion at low concentrations and inhibition at high concentrations. D. rerio embryo culture and MTY assays on embryonic cell proliferation further demonstrate the dosage dependent - cell nutrition and cytotoxicity of the four amino acid neurotransmitters. This experiment suggests that it is necessary to set up a multifaceted D. rerio system covering cell, embryo, larvae and adult, which can effectively ,screen the regeneration -related small molecule compounds and their suitable concentrations.

关 键 词：神经递质 牛磺酸 谷氨酸 甘氨酸 Γ-氨基丁酸 斑马鱼(Danio rerio) 组织再生 

分 类 号：S942.1[农业科学—水产养殖]