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*整体原位杂交法研究斑马鱼(Danio rerio)胚胎发育过程中胸苷酸合成酶(TS)的表达*
牛荣丽, 梁丽英
华侨大学分子药物学研究所教育部分子药物工程中心

摘要:

收集不同发育时期的斑马鱼胚胎, 制备 DIG 标记的 TS 反义 RNA 探针, 采用整体原位杂交方法研究胸苷酸合成酶(TS)基因在斑马鱼胚胎发育各期的时空表达状况。结果表明, 在所取样的各个阶段, TS 基因均有转录, 但其部位不同, 中囊胚过渡前后 mRNA 显现的部位仅存在于受精卵的动物极, 10hpf 时存在于整个胚胎的外围, 至 24hpf 后集中到头部及其躯干部。112hpf 在心脏部位强表达, 其余部位表达相对较弱。

关键词: 胸苷酸合成酶(TS), 斑马鱼, 反义 RNA 探针, 整体原位杂交

DOI：10.11693/hyhz201101012012

分类号:

基金项目:国家自然科学基金资助项目, 40506031 号; 厦门市科技项目, xm090905 号; 福建省生物医药工程研究生教育创新基地资助, 06070205 号

PROFILING OF TS GENE EXPRESSION OF ZEBRAFISH DANIO RERIO EMBRYOGENESIS USING WHOLE MOUNT IN SITU HYBRIDIZATION

NIU Rong-Li, LIANG Li-Ying

Engineering Research Center of Molecular Medicine, Ministry of Education, Institute of Molecular Medicine, Huaqiao University

Abstract:

To analyze the expression pattern of thymidylate synthase (TS) of zebrafish Danio rerio embryogenesis, we collected zebrafish embryos at different phases and prepared TS digoxin-labeled RNA probe using whole mount in situ hybridization. We found that TS expression appeared at early zygote period, continued throughout the entire embryogenesis periods. Expression of TS was also discovered in different parts of the embryo: in the animal pole of the middle gastrula, the margin of whole embryo after 10 hours, in the head and somite after 24 hours post fertilization. After 112 hours post-fertilization, strong TS expression only in the heart. Therefore, TS plays important role in the regulation of zebrafish embryonic development.

Key words: Thymidylate synthase (TS), Zebrafish Danio rerio, RNA antisense probe, Whole mount in situ hybridization

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