Department of Immunology, Genetics and Pathology, Uppsala University, S-751 08, Uppsala, Sweden.
Biol Open. 2022 Mar 15;11(3). doi: 10.1242/bio.059167. Epub 2022 Mar 22.
Fictive locomotion is frequently used to study locomotor output in paralyzed animals. We have evaluated the character of swim episodes elicited by different strategies in zebrafish. Motor output was measured on both sides of a body segment using electrodes and a pipeline for synchronizing stimulation and recording, denoising data and peak-finding was developed. The optomotor response generated swims most equivalent to spontaneous activity, while electrical stimulation and NMDA application caused various artefacts. Our optimal settings, optomotor stimulation using 5-day-old larvae, were combined with calcium imaging and optogenetics to validate the setup's utility. Expression of GCaMP5G by the mnx1 promoter allowed correlation of calcium traces of dozens of motor neurons to the fictive locomotor output. Activation of motor neurons through channelrhodopsin produced aberrant locomotor episodes. This strategy can be used to investigate novel neuronal populations in a high-throughput manner to reveal their role in shaping motor output. This article has an associated First Person interview with the first author of the paper.
虚构运动常被用于研究瘫痪动物的运动输出。我们评估了不同策略引发的斑马鱼游泳片段的特征。使用电极和一个用于同步刺激和记录、数据去噪和峰值检测的管道，在身体节段的两侧测量运动输出。光刺激产生的游泳最接近自发活动，而电刺激和 NMDA 应用会导致各种伪迹。我们的最佳设置是使用 5 天大的幼虫进行光运动刺激，同时结合钙成像和光遗传学来验证设置的实用性。通过 mnx1 启动子表达 GCaMP5G，使得数十个运动神经元的钙迹与虚构的运动输出相关联。通过通道视紫红质激活运动神经元会产生异常的运动片段。这种策略可用于高通量地研究新的神经元群体，以揭示它们在塑造运动输出中的作用。本文有一篇与第一作者的第一人称访谈。
