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This book merges approaches in understanding the function of the light-gated ion channels known as channelrhodopsin together with methods addressing how channelrhodopsins can be used to address biomedical questions on a cellular or organismal level. Since the first molecular identification of channelrhodopsins, a broad range of tools have been created and new approaches developed to both better understand the molecular determinants of channelrhodopsin function as well as to use these and homologous proteins from a variety of species as tools to better understand physiological processes, which this volume addresses. Additionally, channelrhodopsins have become instrumental as a potential treatment for disease states. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Channelrhodopsin: Methods and Protocols provides a resource for those interested in honing their current expertise in this vital area of study as well as potentially branching out into new directions.
List of contents
Molecular Dynamics Simulations of Channelrhodopsin Chimera, C1C2.- Computing Potential of the Mean Force Profiles for Ion Permeation through Channelrhodopsin Chimera, C1C2.- Nanodisc Reconstitution of Channelrhodopsins Heterologously Expressed in Pichia pastoris for Biophysical Investigations.- Characterizing Channelrhodopsin Channel Properties via Two-Electrode Voltage Clamp and Kinetic Modeling.- Charge Transport by Light-Activated Rhodopsins Determined by Electrophysiological Recordings.- Probing Channelrhodopsin Electrical Activity in Algal Cell Populations.- Two-Photon Optogenetic Stimulation of Drosophila Neurons.- Probing Synaptic Signaling with Optogenetic Stimulation and Genetically-Encoded Calcium Reporters.- Optogenetics to Interrogate Neuron-Glia Interactions in Pups and Adults.- Chronic Optogenetic Pacing of Human Induced Pluripotent Stem Cell-Derived Engineered Cardiac Tissues.- Patterned Optogenetic Stimulation Using a DMD-Projector.- Selecting ChannelrhodopsinConstructs for Optimal Visual Restoration in Differing Light Conditions.- Recording Channelrhodopsin-Evoked Field Potentials and Startle Responses from Larval Zebrafish.- Automated Functional Screening for Modulators of Optogenetically-Activated Neural Responses in Living Organisms.- Optogenetic Interrogation of ChR2-Expressing GABAergic Interneurons after Transplantation into the Mouse Brain.- Application of Targeting-Optimized Chronos for Stimulation of the Auditory Pathway.- Channelrhodopsins for Cell-Type Specific Illumination of Cardiac Electrophysiology.- Optogenetic Control of Cardiac Autonomic Neurons in Transgenic Mice.- Dissecting Mechanisms of Motivation within the Nucleus Accumbens Using Optogenetics.- Optogenetic Stimulation of the Central Amygdala Using Channelrhodopsin.- Optical Manipulation of Perfused Mouse Heart Expressing Channelrhodopsin-2 in Rhythm Control.- Chronic Optogenetic Stimulation in Freely-Moving Rodents.
