Module 7C: Supporting Resources on Technical Considerations
The articles below were selected by Scott Owen, Chris Chen, Julia Lemos, and Shana Augustin, faculty from Module 7 of SfN’s Optogenetics Training Series. These resources supplement their presentations, Selecting Appropriate Stimulus Parameters for Optogenetic Manipulations and Validating Optogenetic Experiments.
Use these resources to optimize, validate, and troubleshoot your optogenetic experiments in order to avoid and address potential confounds.
Raimondo et al. describe how chloride pumps (halorhodopsin) can interfere with inhibitory synaptic transmission by affecting GABAA receptor reversal potentials and lead to neuronal excitation via GABAA receptors.
In this 2016 paper, Mahn et al. demonstrate that terminal inactivation with both archaerhodopsin and halorhodopsin attenuates evoked neurotransmitter release, whereas prolonged inactivation with archearhodopsin alone increases spontaneous release. These findings reveal that the biophysical properties of presynaptic terminals dictate unique conditions for optogenetic manipulation.
El-Gaby et al. show archaerhodopsin (Arch)-induced inactivation of axon terminals is mediated by intracellular pH control. This pH-mediated synaptic silencing is selective and reversible.
Mattis et al. provide a direct comparison of the activation and inactivation kinetics between many microbial opsins under matched experimental conditions.
Stujenske, Spellman, and Gordon provide an estimation of heat generated by light delivery through an optical fiber and show heat generated by light stimulation can alter the firing of neurons, even in the absence of opsin expression.
Jackman et al. compare different viral vectors and opsin expression methods to optimize channelrhodopsin-2-evoked presynaptic activation at high stimulation frequencies.
Harris, Wo, and Zheng present two protocols for stereotaxic-guided placement of recombinant viral vectors into the live mouse brain.
This 2012 resource by Madisen et al. presents a toolbox of four knock-in mouse lines engineered for Cre-dependent expression of bacterial opsins for robust activation or silencing of neuronal activity.
Watakabe et al. provide a comparative analysis of cortical viral spread in marmoset, mouse, and macaque cerebral cortex using different adeno-associated virus serotypes and promoters.
Pillay et al. provide information about different AAV serotypes and their distinct interactions with the AAV receptor. These interactions have important implications for AAV vector transduction efficacy and tropism.
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