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斑马鱼幼体的三氯乙酸固定及抗体染色

Trichloroacetic Acid Fixation and Antibody Staining of Zebrafish Larvae.

作者信息

Martin E Anne, Ijaz Sundas, Pereda Alberto E, Miller Adam C

机构信息

Institute of Neuroscience, University of Oregon, Eugene, United States.

Dominick P. Purpura Department of Neuroscience, Albert Einstein College of Medicine, Bronx, United States.

出版信息

Bio Protoc. 2022 Jan 20;12(2):e4289. doi: 10.21769/BioProtoc.4289.

Abstract

Larval zebrafish have been established as an excellent model for examining vertebrate biology, with many researchers using the system for neuroscience. Controlling a fast escape response of the fish, the Mauthner cells and their associated network are an attractive model, given their experimental accessibility and fast development, driving ethologically relevant behavior in the first five days of development. Here, we describe methods for immunostaining electrical and chemical synapse proteins at 3-7 days post fertilization (dpf) in zebrafish using tricholoracetic acid fixation. The methods presented are ideally suited to easily visualize neural circuits and synapses within the fish.

摘要

斑马鱼幼体已成为研究脊椎动物生物学的优秀模型,许多研究人员将该系统用于神经科学研究。鉴于Mauthner细胞及其相关网络在实验上易于操作且发育迅速,能在发育的前五天驱动与行为学相关的行为,它们在控制鱼类快速逃避反应方面是一个有吸引力的模型。在此,我们描述了在受精后3至7天(dpf)使用三氯乙酸固定法对斑马鱼电突触和化学突触蛋白进行免疫染色的方法。所介绍的方法非常适合于轻松可视化鱼体内的神经回路和突触。

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