Friedrich Miescher Institute for Biomedical Research Basel, Switzerland ; University of Basel Basel, Switzerland.
Friedrich Miescher Institute for Biomedical Research Basel, Switzerland ; Institut Universitaire en Santé Mentale de Québec Québec, QC, Canada ; Département de Biochimie, Microbiologie et Bio-informatique, Université Laval Québec, QC, Canada.
Front Neural Circuits. 2014 May 6;8:41. doi: 10.3389/fncir.2014.00041. eCollection 2014.
The zebrafish has various advantages as a model organism to analyze the structure and function of neural circuits but efficient viruses or other tools for fast gene transfer are lacking. We show that transgenes can be introduced directly into the adult zebrafish brain by herpes simplex type I viruses (HSV-1) or electroporation. We developed a new procedure to target electroporation to defined brain areas and identified promoters that produced strong long-term expression. The fast workflow of electroporation was exploited to express multiple channelrhodopsin-2 variants and genetically encoded calcium indicators in telencephalic neurons for measurements of neuronal activity and synaptic connectivity. The results demonstrate that HSV-1 and targeted electroporation are efficient tools for gene delivery into the zebrafish brain, similar to adeno-associated viruses and lentiviruses in other species. These methods fill an important gap in the spectrum of molecular tools for zebrafish and are likely to have a wide range of applications.
斑马鱼作为分析神经回路结构和功能的模式生物具有多种优势，但缺乏有效的病毒或其他快速基因转移工具。我们表明，单纯疱疹 I 型病毒 (HSV-1) 或电穿孔可将转基因直接引入成年斑马鱼大脑。我们开发了一种将电穿孔靶向特定脑区的新程序，并确定了产生强长期表达的启动子。电穿孔的快速工作流程被用于在端脑神经元中表达多种通道视紫红质-2 变体和遗传编码的钙指示剂，以测量神经元活动和突触连接。结果表明，HSV-1 和靶向电穿孔是将基因递送入斑马鱼大脑的有效工具，类似于其他物种中的腺相关病毒和慢病毒。这些方法填补了斑马鱼分子工具谱中的一个重要空白，可能具有广泛的应用。
