A new platform for long-term tracking and recording of neural activity and simultaneous optogenetic control in freely behaving Caenorhabditis elegans

Journal of Neuroscience Methods
Keiko Gengyo-AndoJunichi Nakai

Abstract

Real-time recording and manipulation of neural activity in freely behaving animals can greatly advance our understanding of how neural circuits regulate behavior. Ca2+imaging and optogenetic manipulation with optical probes are key technologies for this purpose. However, integrating the two optical approaches with behavioral analysis has been technically challenging. Here, we developed a new imaging system, ICaST (Integrated platform for Ca2+imaging, Stimulation, and Tracking), which combines an automatic worm tracking system and a fast-scanning laser confocal microscope, to image neurons of interest in freely behaving C. elegans. We optimized different excitation wavelengths for the concurrent use of channelrhodopsin-2 and G-CaMP, a green fluorescent protein (GFP)-based, genetically encoded Ca2+indicator. Using ICaST in conjunction with an improved G-CaMP7, we successfully achieved long-term tracking and Ca2+imaging of the AVA backward command interneurons while tracking the head of a moving animal. We also performed all-optical manipulation and simultaneous recording of Ca2+dynamics from GABAergic motor neurons in conjunction with behavior monitoring. Our system differs from conventional systems in that it does not require fl...Continue Reading

Citations

Jun 17, 2020·Journal of Neurogenetics·Inka BusackHenrik Bringmann
Jul 9, 2021·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Haiwen LiHeng Mao
Oct 30, 2021·ELife·Frank K LeeMichael I Kotlikoff

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