摘要
目的构建α细胞细胞核特异表达红色荧光蛋白的转基因鱼,为详细研究斑马鱼胰岛α细胞的功能和药物筛选奠定基础。方法利用In-Fusion克隆、I-SecI重组、显微注射等技术构建了斑马鱼α细胞细胞核表达红色荧光的稳定遗传转基因品系Tg(-2.8gcga:H2BmCherry)。结果建立并筛选获得标记胰岛α细胞细胞核表达红色荧光的转基因斑马鱼,该品系能准确标记所有的斑马鱼胰岛α细胞,其细胞在共聚焦显微镜下清晰可见,细胞间轮廓分别,比已有的转基因品系的α细胞的分辨率有显著提高。结论成功构建了新的胰岛α细胞细胞核红色荧光标记的转基因鱼,可用于后期在整体动物水平上研究胰岛α细胞的发育与功能,及开展药物筛选;为进一步对胰岛α细胞的深入研究提供了一个便捷又直观的新型工具。
Objective To develop a transgenic zebrafish which target expression of m Cherry in α cells nuclei,in order to study the pancreatic α cells regulation of glucose homeostasis and diabetes pathophysiology using zebrafish as model.Methods With combination of In-Fusion cloning,I-Sce I meganuclease-mediated transgenesis,and microinjection techniques,a stable transgenic zebrafish line named Tg(-2.8 gcga: H2 BmCherry) was generated,which expressing m Cherry fluorescence protein in the nuclei of zebrafish α cells.Results The stable line of Tg(-2.8 gcga: H2 BmCherry),which α cells nuclei clearly labeled with m Cherry was obtained.All the m Cherry expressed cells in the transgenic line were the glucagonexpressing cells.The resolution of m Cherry fluorescence was high,and individual α cell was easy to count.Conclusion A transgenic zebrafish with α cells nuclei specifically expressed of m Cherry was obtained in this study.It could as useful tools for studies include α-cells development,α-cells physiology,and even anti-diabetes drug screening.
作者
林春雨
郏建新
康琪
吴志强
李明玉
LIN Chunyu;JIA Jianxin;KANG Qi;WU Zhiqiang;LI Mingyu(College of Marine Life Sciences,Ocean University of China,Qingdao 266003,China;School of Pharmaceutical Sciences,Xiamen University,Xiamen 361102,China)
出处
《药学研究》
CAS
2020年第5期256-260,265,共6页
Journal of Pharmaceutical Research
基金
国家自然科学基金(No.81670709)
福建省自然科学基金(No.2017J01145)
中央高校基本科研业务费(No.20720170104、20720180044)。
