Abstract
With the progress of optogenetics, the activities of genetically identified neurons can be optically silenced to determine whether the neurons in question are necessary for the network performance of the behavioral expression. This logical induction is expected to be improved by the application of the Na+ pump rhodopsins (NaRs), which hyperpolarize the membrane potential with negligible influence on the ionic/pH balance. Here, we made several chimeric NaRs between two NaRs, KR2 and IaNaR from Krokinobacter eikastus and Indibacter alkaliphilus, respectively. We found that one of these chimeras, named I1K6NaR, exhibited some improvements in the membrane targeting and photocurrent properties over native NaRs. The I1K6NaR-expressing cortical neurons were stably silenced by green light irradiation for a certain long duration. With its rapid kinetics and voltage dependency, the photoactivation of I1K6NaR would specifically counteract the generation of action potentials with less hyperpolarization of the neuronal membrane potential than KR2.
Highlights
A neural system is a kind of computing system comprised by a vast number of neurons
To study the ion-transport selectivity of IaNaR, the protein was expressed in E. coli cells and the light-induced pH change was monitored upon illumination of the cell suspension
This result indicates that the increase in pH represents the secondary H+-uptake into the E. coli cell body to compensate the hyperpolarized membrane potential generated by the transport of ions other than H+ by IaNaR
Summary
A neural system is a kind of computing system comprised by a vast number of neurons. In recent years the optical methods have been developed to manipulate genetically targeted neurons using light under precise spatiotemporal resolution [1,2,3,4]. The neuronal excitability is enhanced by light if light-sensitive cation channels such as channelrhodopsin-2 (ChR2) are expressed in a neuron [5,6]. The resulting neural network or behavior responses suggest that the stimulated neuron is involved in the responses as a sufficient condition, it is necessary to avoid oversimplification [4]. Optical silencing of the neural activity could determine whether the neuron in question is necessary for the network performance of the behavioral expression
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