Image credit: Martin420 (CC BY-SA 4.0)
One way to study the role of a specific neuron is to activate or inhibit the cell and then observe the consequences. This can be achieved by using optogenetics, a technique that involves introducing ‘light-gated’ ion channels in the outer membrane of a target neuron. When light is shone on the cell, these pore-like proteins open their channels: this allows ions to move into or out of the neuron.
Ions flow from high concentration to low concentration areas. Typically, when a neuron is at rest, there are fewer chloride ions inside the cell than outside. Activating a light-gated chloride channel should thus cause these negatively charged ions to enter the neuron. The charge inside of the cell would become more negative relative to the outside: this would inhibit the neuron, making it less likely to fire.
Here, Messier et al. looked into using a light-gated chlorine channel called GtACR2 to inhibit the activity of neurons in mouse brain slices, but the results were not as expected. Activating the chloride channel did inhibit the cell body, the area of the neuron that contains the nucleus. Yet, it had the opposite effect in the axon, the structure that carries electrical signals away from the cell body.
There, activating GtACR2 caused chloride ions to leave the axon, which resulted in the neuron firing. Testing other types of optogenetic chloride channels produced the same result. Further experiments revealed that the concentration of chloride ions is higher inside the axon than the cell body, explaining the observed effects.
Messier et al. then tried to redistribute the channels from the axon to the cell body, where the proteins are inhibitory. This was accomplished by fitting GtACR2 with a molecular tag that acts like an address label, with the cell body as the target destination. Overall, when these modified channels were activated, the neuron was more strongly inhibited.
Ultimately, the GtACR2 channel designed by Messier et al. is a powerful new inhibitory optogenetic tool. In addition, this tool could be used to study chloride gradients in brain regions, cell types and areas of cells that are otherwise difficult to access.
