In this presentation, faculty cover some of the key technical issues that arise when designing and interpreting optogenetic experiments. Specifically, Chris Chen and Scott Owen will:

• Describe how to select optimal tools and stimulation parameters for an optogenetics experiment.
• Identify potential confounds related to stimulus parameters.
• Describe key methods to validate optogenetic tools.

After watching this presentation, you should be able to better understand how to select optimal tools and stimulation parameters for an optogenetics experiment.

Ask Module 7 Faculty Your Questions

SfN members have the opportunity to submit questions from August 1 – August 16 to Chris Chen, Scott Owen, Julia Lemos, and Shana Augustin in a Neuronline Community discussion thread. They will answer your questions live on Thursday, August 16, from 1–2 p.m. EDT, so make sure to visit the Neuronline Community forum then.

Visit the Community forum for all eight modules to share your insights and best practices, ask questions, and engage with other training series’ participants.

Speakers

Christopher Chen, PhD

Christopher Chen is a postdoctoral fellow in the department of neurobiology at Harvard Medical School. His research focuses on identifying and describing novel outputs of the cerebellum. Chen earned his PhD at Albert Einstein College of Medicine.

Scott Owen, PhD

Scott Owen is a staff scientist and completing his postdoctoral training at the Gladstone Institute of Neurological Disease at the University of California, San Francisco. His research focuses on how local microcircuits facilitate the storage and processing of information and how defects in inhibitory microcircuits can underlie neurological and psychiatric disease. Owen earned his PhD in molecular and cellular physiology from Stanford University.

In this presentation, faculty will provide information to optimize, validate, and troubleshoot optogenetics experiments in order to avoid and address potential confounds. Specifically, Julia Lemos and Shana Augustin will:

• Describe how to achieve optimal opsin expression.
• Describe how to design control experiments to optimize opsin expression while limiting confounds.
• Describe anatomical and functional validation of opsin expression.

After watching this presentation, you should be able to identify and understand how to address common potential pitfalls in optogenetics experiments.

Ask Module 7 Faculty Your Questions

SfN members have the opportunity to submit questions from August 1 – August 16 to Chris Chen, Scott Owen, Julia Lemos, and Shana Augustin in a Neuronline Community discussion thread. They will answer your questions live on Thursday, August 16, from 1–2 p.m. EDT, so make sure to visit the Neuronline Community forum then.

Visit the Community forum for all eight modules to share your insights and best practices, ask questions, and engage with other training series’ participants.

Speakers

Julia Lemos, PhD

Julia Lemos is an assistant professor in the department of neuroscience and a member of the Medical Discovery Team on Addiction at the University of Minnesota, Twin Cities. Her research focuses on understanding how acute and chronic stress effect the function of neural circuits important for motivation and cognition. Lemos is particularly interested in the role of neuropeptides in these processes. She earned her PhD in neurobiology and behavior from the University of Washington and did her postdoctoral fellowship at the National Institute on Alcoholism and Alcohol Abuse.

Shana Augustin, PhD

Shana Augustin is a senior NIH Intramural Research Training Award postdoctoral fellow in the Laboratory for Integrative Neuroscience at the National Institute on Alcohol Abuse and Alcoholism. Her research focuses on understanding the cellular mechanisms for synaptic and circuit level plasticity that underlie learning and motor control using a variety of approaches, including mouse genetics, electrophysiology, pharmacology, fast-scan cyclic voltammetry, photometry, and behavior. After earning her BS in biology at the University of Virgin Islands in St. Thomas, United States Virgin Islands, Augustin completed her PhD in neurobiology at the University of Chicago.

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.

Optogenetic Silencing Strategies Differ in Their Effects on Inhibitory Synaptic Transmission

Raimondo et al. describe how chloride pumps (halorhodopsin) can interfere with inhibitory synaptic transmission by affecting GABA_A receptor reversal potentials and lead to neuronal excitation via GABA_A receptors.

Biophysical Constraints of Optogenetic Inhibition at Presynaptic Terminals

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.

Archaerhodopsin Selectively and Reversibly Silences Synaptic Transmission Through Altered pH

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.

Principles for Applying Optogenetic Tools Derived From Direct Comparative Analysis of Microbial Opsins

Mattis et al. provide a direct comparison of the activation and inactivation kinetics between many microbial opsins under matched experimental conditions.

Modeling the Spatiotemporal Dynamics of Light and Heat Propagation for In Vivo Optogenetics

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.

Achieving High-Frequency Optical Control of Synaptic Transmission

Jackman et al. compare different viral vectors and opsin expression methods to optimize channelrhodopsin-2-evoked presynaptic activation at high stimulation frequencies.

Adeno‐Associated Viral Vectors for Anterograde Axonal Tracing With Fluorescent Proteins in Nontransgenic and Cre Driver Mice

Harris, Wo, and Zheng present two protocols for stereotaxic-guided placement of recombinant viral vectors into the live mouse brain.

A Toolbox of Cre-Dependent Optogenetic Transgenic Mice for Light-Induced Activation and Silencing

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.  

Comparative Analyses of Adeno-Associated Viral Vector Serotypes 1, 2, 5, 8 and 9 in Marmoset, Mouse and Macaque Cerebral Cortex

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.

Adeno-Associated Virus (AAV) Serotypes Have Distinctive Interactions With Domains of the Cellular AAV Receptor

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.

Visit the Community forum for all eight modules to share your insights and best practices, ask questions, and engage with other training series’ participants.

Complete this short survey to provide valuable feedback about Module 7 to SfN and the series faculty.

To design and conduct a successful and rigorous optogenetics experiment, there are a few key technical issues that must be considered. These include achieving optimal opsin expression, selecting appropriate stimulation parameters, and validating opsin expression and function.

SfN members have the opportunity to submit questions from August 1 – August 16 to Module 7 faculty — Chris Chen, Scott Owen, Julia Lemos, and Shana Augustin — in a Neuronline Community discussion thread. They will answer your questions live on Thursday, August 16, from 1–2 pm EDT, so make sure to visit the Neuronline Community forum then.

Visit the Community forum for all eight modules to share your insights and best practices, ask questions, and engage with other training series’ participants.

Complete this short survey to provide valuable feedback about Module 7 to SfN and the series faculty.

Speakers

Scott Owen, PhD

Scott Owen is a staff scientist and completing his postdoctoral training at the Gladstone Institute of Neurological Disease at the University of California, San Francisco. His research focuses on how local microcircuits facilitate the storage and processing of information and how defects in inhibitory microcircuits can underlie neurological and psychiatric disease. Owen earned his PhD in molecular and cellular physiology from Stanford University.

Christopher Chen, PhD

Christopher Chen is a postdoctoral fellow in the department of neurobiology at Harvard Medical School. His research focuses on identifying and describing novel outputs of the cerebellum. Chen earned his PhD at Albert Einstein College of Medicine.

Julia Lemos, PhD

Julia Lemos is an assistant professor in the department of neuroscience and a member of the Medical Discovery Team on Addiction at the University of Minnesota, Twin Cities. Her research focuses on understanding how acute and chronic stress effect the function of neural circuits important for motivation and cognition. Lemos is particularly interested in the role of neuropeptides in these processes. She earned her PhD in neurobiology and behavior from the University of Washington and did her postdoctoral fellowship at the National Institute on Alcoholism and Alcohol Abuse.

Shana Augustin, PhD

Shana Augustin is a senior NIH Intramural Research Training Award postdoctoral fellow in the Laboratory for Integrative Neuroscience at the National Institute on Alcohol Abuse and Alcoholism. Her research focuses on understanding the cellular mechanisms for synaptic and circuit level plasticity that underlie learning and motor control using a variety of approaches, including mouse genetics, electrophysiology, pharmacology, fast-scan cyclic voltammetry, photometry, and behavior. After earning her BS in biology at the University of Virgin Islands in St. Thomas, United States Virgin Islands, Augustin completed her PhD in neurobiology at the University of Chicago.