利用双光子激光消融斑马鱼幼虫中的神经元群体及其通过钙成像和行为记录进行评估
Ablation of a Neuronal Population Using a Two-photon Laser and Its Assessment Using Calcium Imaging and Behavioral Recording in Zebrafish Larvae.
作者信息
Muto Akira, Kawakami Koichi
机构信息
Division of Molecular and Developmental Biology, National Institute of Genetics, Department of Genetics, SOKENDAI (The Graduate University for Advanced Studies);
Division of Molecular and Developmental Biology, National Institute of Genetics, Department of Genetics, SOKENDAI (The Graduate University for Advanced Studies).
出版信息
J Vis Exp. 2018 Jun 2(136):57485. doi: 10.3791/57485.
To identify the role of a subpopulation of neurons in behavior, it is essential to test the consequences of blocking its activity in living animals. Laser ablation of neurons is an effective method for this purpose when neurons are selectively labeled with fluorescent probes. In the present study, protocols for laser ablating a subpopulation of neurons using a two-photon microscope and testing of its functional and behavioral consequences are described. In this study, prey capture behavior in zebrafish larvae is used as a study model. The pretecto-hypothalamic circuit is known to underlie this visually-driven prey catching behavior. Zebrafish pretectum were laser-ablated, and neuronal activity in the inferior lobe of the hypothalamus (ILH; the target of the pretectal projection) was examined. Prey capture behavior after pretectal ablation was also tested.
为了确定神经元亚群在行为中的作用，在活体动物中测试阻断其活动的后果至关重要。当神经元用荧光探针进行选择性标记时，激光消融神经元是实现这一目的的有效方法。在本研究中，描述了使用双光子显微镜对神经元亚群进行激光消融及其功能和行为后果测试的方案。在这项研究中，斑马鱼幼体的捕食行为被用作研究模型。已知前顶盖-下丘脑回路是这种视觉驱动的捕食行为的基础。对斑马鱼的前顶盖进行激光消融，并检查下丘脑下叶（ILH；顶盖前投射的目标）中的神经元活动。还测试了顶盖前消融后的捕食行为。
Zotero 插件