Control and characterization of memory function

Abstract

Provided herein are devices and methods for reversibly controlling memory function in living non-human animals. Some variations of methods for affecting memory function comprise temporarily inhibiting neurons of the hippocampus (e.g., neurons of the dorsal CA1 field of the hippocampus) during the acquisition or retrieval of a memory. Alternatively or additionally, methods for reversibly affecting memory function comprise inhibiting neurons of the amygdala (e.g. basolateral amygdala) and/or neurons of the cingulate cortex (e.g., anterior cingulated cortex). Methods for disrupting the formation and recall of memories by inhibiting excitatory neurons expressing light-activated proteins are disclosed herein. One or more methods for reversibly affecting memory function described herein can be used to evaluate the effectiveness of pharmacological agents in treating PTSD and/or various memory disorders.

1 . A light-activated protein expressed on the cell membrane of excitatory neurons in the dorsal CA1 field of the hippocampus, the anterior cingulated cortex, or the basolateral amygdala of an animal, wherein the protein is responsive to light and is capable of inhibiting depolarization of the neurons when the neurons are illuminated with the light, wherein the illumination of the protein reversibly affects memory function. 
     
     
         2 .- 3 . (canceled) 
     
     
         4 . The non-human animal of  claim 1 , wherein the memory function that is affected is memory retrieval or memory formation. 
     
     
         5 . The non-human animal of  claim 1 , wherein the memory is a fearful memory. 
     
     
         6 . The non-human animal of  claim 1 , wherein the memory is a remote memory. 
     
     
         7 . The non-human animal of  claim 1 , wherein the protein is selected from the group consisting of NpHR, BR, AR, and GtR3. 
     
     
         8 . The non-human animal of  claim 7 , wherein the protein comprises an amino acid sequence at least 95% identical to the sequence shown in SEQ ID NO:3. 
     
     
         9 . The non-human animal of  claim 8 , wherein the protein further comprises an endoplasmic reticulum (ER) export signal and/or a membrane trafficking signal. 
     
     
         10 . The non-human animal of  claim 8 , wherein the amino acid sequence is linked to the ER export signal through a linker. 
     
     
         11 . The non-human animal of  claim 9 , wherein the ER export signal comprises the amino acid sequence FCYENEV. 
     
     
         12 . The non-human animal of  claim 9 , wherein the membrane trafficking signal comprises the amino acid sequence KSRITSEGEYIPLDQIDINV. 
     
     
         13 . The non-human animal of  claim 7 , wherein the protein comprises the amino acid sequence shown in SEQ ID NO:5 or SEQ ID NO:6. 
     
     
         14 . A brain tissue slice comprising a brain region selected from the group consisting of the dorsal CA1 field of the hippocampus, the basolateral amygdala, and the anterior cingulated cortex, wherein a light-activated protein is expressed on the cell membrane of excitatory neurons of the brain region, wherein the protein is responsive to light and is capable of inhibiting depolarization of the neurons when the neurons are illuminated with the light, wherein the illumination of the protein reversibly affects memory function. 
     
     
         15 . A method for reversibly affecting memory retrieval or formation in an individual comprising:
 administering a polynucleotide encoding a light-activated protein to the dorsal CA1 field of the hippocampus, the anterior cingulated cortex, or the basolateral amygdala in the individual, wherein light-activated protein is expressed on the cell membrane of the excitatory neurons in the dorsal CA1 field of the hippocampus, the anterior cingulated cortex, or the basolateral amygdala, and the protein is responsive to light and is capable of inhibiting depolarization of the neurons when the neurons are illuminated with the light, whereby activating the protein by the light reversibly affects memory retrieval or formation of an event in the individual.   
     
     
         16 .- 17 . (canceled) 
     
     
         18 . The method of  claim 15 , wherein the polynucleotide is a vector. 
     
     
         19 . The method of  claim 18 , wherein the vector is a viral vector selected from the group consisting of an AAV vector, a retroviral vector, an adenoviral vector, an HSV vector, and a lentiviral vector. 
     
     
         20 . A method for reversibly affecting memory retrieval or formation comprising:
 inhibiting depolarization of excitatory neurons in the dorsal CA1 field of the hippocampus, the anterior cingulated cortex, or the basolateral amygdala during memory retrieval or formation of an event in an individual, wherein a light-activated protein is expressed on the cell membrane of the excitatory neurons in the dorsal CA1 field of the hippocampus, the anterior cingulated cortex, or the basolateral amygdala of the individual, wherein the protein is responsive to light and is capable of inhibiting depolarization of the neurons when the neurons are illuminated with the light.   
     
     
         21 .- 22 . (canceled) 
     
     
         23 . The method of  claim 15 , wherein the event is a fearful event. 
     
     
         24 . The method of  claim 15 , wherein the individual is a human. 
     
     
         25 . The method of  claim 15 , wherein the individual is a non-human animal. 
     
     
         26 . A method for treating post-traumatic stress disorder in an individual comprising:
 administering a polynucleotide encoding a light-activated protein to the dorsal CA1 field of the hippocampus, the anterior cingulated cortex, or the basolateral amygdala in the individual, wherein light-activated protein is expressed on the cell membrane of the excitatory neurons in the dorsal CA1 field of the hippocampus, the anterior cingulated cortex, or the basolateral amygdala and the protein is responsive to light and is capable of inhibiting depolarization of the neurons when the neurons are illuminated with the light, whereby activating the protein by the light reversibly affects memory retrieval or formation of an event in the individual.   
     
     
         27 . (canceled) 
     
     
         28 . The method of  claim 26 , wherein the polynucleotide is a vector. 
     
     
         29 . The method of  claim 28 , wherein the vector is a viral vector selected from the group consisting of an AAV vector, a retroviral vector, an adenoviral vector, an HSV vector, and a lentiviral vector. 
     
     
         30 . A method of screening a pharmacological agent that affects memory retrieval or formation comprising:
 a) contacting excitatory neurons in the dorsal CA1 field of the hippocampus, the anterior cingulated cortex, or the basolateral amygdala during memory retrieval or formation of an event in a non-human animal with a pharmacological agent, wherein the non-human animal comprises a light-activated protein expressed on the cell membrane of excitatory neurons in the dorsal CA1 field of the hippocampus, the anterior cingulated cortex, or the basolateral amygdala of the animal, wherein the protein is responsive to light and is capable of inhibiting depolarization of the neurons when the neurons are illuminated with the light;   b) inhibiting depolarization of the excitatory neurons in the dorsal CA1 field of the hippocampus, the anterior cingulated cortex, or the basolateral amygdala during memory retrieval or formation of an event; and   c) determining if the pharmacological agent affects memory retrieval or formation in the presence or absence of the light.   
     
     
         31 .- 32 . (canceled) 
     
     
         33 . The method of  claim 15 , wherein the protein is selected from the group consisting of NpHR, BR, AR, and GtR3. 
     
     
         34 . The method of  claim 33 , wherein the NpHR protein comprises an amino acid sequence at least 95% identical to the sequence shown in SEQ ID NO:3. 
     
     
         35 . The method of  claim 34  wherein the NpHR protein further comprises an endoplasmic reticulum (ER) export signal and/or a membrane trafficking signal. 
     
     
         36 . The method of  claim 35 , wherein the amino acid sequence at least 95% identical to the sequence shown in SEQ ID NO:3 is linked to the ER export signal and/or the membrane trafficking signal through a linker. 
     
     
         37 . The method of  claim 35 , wherein the ER export signal comprises the amino acid sequence FCYENEV. 
     
     
         38 . The method of  claim 35 , wherein the membrane trafficking signal comprise the amino acid sequence KSRITSEGEYIPLDQIDINV. 
     
     
         39 . The method of  claim 33 , wherein the NpHR protein comprises the amino acid sequence shown in SEQ ID NO:5 or SEQ ID NO:6.