Stabilized step function opsin proteins and methods of using the same
Provided herein are compositions comprising non-human animals comprising neurons expressing stabilized step function opsin proteins on neural plasma membranes and methods of using the same to selectively depolarize neurons residing in microcircuits of the pre-frontal cortex to affect one or more social behaviors, communications, and/or conditioned behaviors in the non-human animal.
1. A method for identifying a chemical compound that restores a social behavior, communication, and/or conditioned behavior in a non-human mammal, the method comprising:
(a) depolarizing excitatory neurons in the prefrontal cortex of the non-human mammal, wherein the excitatory neurons of the prefrontal cortex are genetically modified to express a light-activated cation channel protein on the cell membrane of the excitatory neurons, wherein depolarization comprises activating the light-activated cation channel protein with light, and wherein the non-human mammal is a mouse or a rat,
wherein the light-activated cation channel protein comprises an amino acid sequence at least 85% identical to the amino acid sequence depicted in one of SEQ ID NOs:1-4, and comprises amino acid substitutions at amino acid residues corresponding to C128, D156, or both C128 and D156 of the amino acid sequence of SEQ ID NO:1, wherein depolarizing the excitatory neuron inhibits one or more social behaviors, communications, and/or conditioned behaviors in the non-human mammal;
(b) administering a chemical compound to the non-human mammal; and
(c) determining if the administration of the chemical compound to the non-human animal restores said one or more social behaviors, communications, and/or conditioned behaviors in the non-human mammal.
2. The method of claim 1 , wherein the social behavior is selected from the group consisting of: allogrooming, resident-intruder aggression, isolation-induced fighting, sexual behavior, parental behavior, social recognition, and auditory communication.
3. The method of claim 1 , wherein the residue corresponding to C128 of the amino acid sequence of SEQ ID NO:1 is substituted with serine.
4. The method of claim 1 , wherein the residue corresponding to D156 of the amino acid sequence of SEQ ID NO:1 is substituted to alanine.
5. The method of claim 1 , wherein the light-activated cation channel protein comprises an amino acid sequence at least 90% identical to the amino acid sequence depicted in one of SEQ ID NOs:1-4.
6. The method of claim 1 , wherein the light-activated cation channel protein comprises an amino acid sequence at least 95% identical to the amino acid sequence depicted in one of SEQ ID NOs:1-4.
7. The method of claim 1 , wherein the light-activated cation channel protein comprises an amino acid sequence at least 95% identical to the sequence shown in SEQ ID NO:1, and wherein C128 is substituted with serine and D156 is substituted with alanine.
8. The method of claim 1 , wherein the non-human mammal is a mouse.
9. The method of claim 1 , wherein the non-human mammal is a rat.
10. The method of claim 1 , wherein the neurons are in the infralimbic or prelimbic subregions of the medial prefrontal cortex.