Cell type-specific and time-dependent light exposure contribute to silencing in neurons expressing Channelrhodopsin-2

Abstract

Channelrhodopsin-2 (ChR2) has quickly gained popularity as a powerful tool for eliciting genetically targeted neuronal activation. However, little has been reported on the response kinetics of optogenetic stimulation across different neuronal subtypes. With excess stimulation, neurons can be driven into depolarization block, a state where they cease to fire action potentials. Herein, we demonstrate that light-induced depolarization block in neurons expressing ChR2 poses experimental challenges for stable activation of specific cell types and may confound interpretation of experiments when ‘activated’ neurons are in fact being functionally silenced. We show both ex vivo and in vivo that certain neuronal subtypes targeted for ChR2 expression become increasingly susceptible to depolarization block as the duration of light pulses are increased. We find that interneuron populations have a greater susceptibility to this effect than principal excitatory neurons, which are more resistant to light-induced depolarization block. Our results highlight the need to empirically determine the photo-response properties of targeted neurons when using ChR2, particularly in studies designed to elicit complex circuit responses in vivo where neuronal activity will not be recorded simultaneous to light stimulation.The brain is a highly complex structure composed of trillions of interconnecting nerve cells. The pattern of connections between these cells gives rise to the various brain circuits that govern how the brain functions. Understanding how the brain is wired together is important for determining how ‘faulty circuits’ contribute to various neurological disorders.New optogenetic technique tools allow neuroscientists to turn on specific neurons simply by shining light on them. These techniques involve genetically manipulating the organisms so that their neurons express proteins that are activated when they are exposed to light of a particular wavelength. However, it is important to understand the limitations of this approach—including the possibility that the light might actually turn off some neurons—when using it to study animal behavior.Now, Herman, Huang et al. show that shining light pulses for long durations onto neurons expressing a light-activated protein called channelrhodopsin-2 causes the neurons to become silenced rather than activated. Moreover, certain types of neurons, called interneurons, are more susceptible to this effect—termed ‘depolarization block’—than the other types of neurons.Researchers need to be mindful of this effect when channelrhodopsin-2 is used in optogenetic experiments to study the behavior of living animals. However, this silencing property could be useful in experiments that investigate situations in which depolarization block is thought to contribute to brain function and health: such as in the treatments of schizophrenia and Parkinson’s disease.