Upconversion of light for use in optogenetic methods

Abstract

Provided herein are compositions comprising lanthanide-doped nanoparticles which upconvert electromagnetic radiation from infrared or near infrared wavelengths into the visible light spectrum. Also provided herein are methods activating light-responsive opsin proteins expressed on plasma membranes of neurons and selectively altering the membrane polarization state of the neurons using the light delivered by the lanthanide-doped nanoparticles.

1 .- 13 . (canceled) 
     
     
         14 . A method to hyperpolarize the plasma membrane of a neural cell in an individual comprising:
 (a) placing a plurality of lanthanide-doped nanoparticles in proximity to the neural cell, wherein the nanoparticles comprise NaYF 4 :Yb/X/Gd, wherein X is erbium (Er), thulium (Tm), or Er/Tm; and   (b) exposing the plurality of nanoparticles to electromagnetic radiation in the infrared (IR) or near infrared (NIR) spectrum, wherein the electromagnetic radiation in the IR or NIR spectrum is upconverted into yellow, amber or red light in the visible spectrum by the nanoparticles, and wherein a light-responsive opsin comprising a light-responsive chloride pump is expressed on the plasma membrane and activation of the opsin by the light in the visible spectrum induces the hyperpolarization of the plasma membrane.   
     
     
         15 . A method to hyperpolarize the plasma membrane of a neural cell in an individual comprising:
 (a) administering a polynucleotide encoding a light-responsive opsin to an individual, wherein the light-responsive opsin comprises a light-responsive chloride pump and is expressed on the plasma membrane of a neural cell in the individual and the opsin is capable of inducing membrane hyperpolarization of the neural cell when illuminated with light;   (b) administering a plurality of lanthanide-doped nanoparticles in proximity to the neural cell, wherein the nanoparticles comprise NaYF 4 :Yb/X/Gd, wherein X is erbium (Er), thulium (Tm), or Er/Tm; and   (c) exposing the plurality of nanoparticles to electromagnetic radiation in the infrared (IR) or near infrared (NIR) spectrum, wherein the electromagnetic radiation in the IR or NIR spectrum is upconverted into yellow, amber or red light in the visible spectrum and the activation of the opsin by the light in the visible spectrum induces the hyperpolarization of the plasma membrane.   
     
     
         16 . The method of  claim 14 , wherein the light-responsive opsin comprises an amino acid sequence having at least 85% amino acid sequence identity to SEQ ID NO:1. 
     
     
         17 . (canceled) 
     
     
         18 . The method of  claim 14 , wherein the electromagnetic energy in the IR or NIR spectrum is upconverted into light having a wavelength of about 580 nm to about 630 nm. 
     
     
         19 . The method of  claim 14 , wherein the electromagnetic energy in the IR or NIR spectrum is upconverted into light having a wavelength of about 630 nm to about 740 nm. 
     
     
         20 . The method of  claim 14 , wherein the electromagnetic energy in the IR or NIR spectrum is upconverted into light having a wavelength corresponding to yellow or amber light. 
     
     
         21 . The method of  claim 14 , wherein the electromagnetic energy in the IR or NIR spectrum is upconverted into light having a wavelength corresponding to red light. 
     
     
         22 . The method of  claim 14 , wherein the individual is a non-human animal. 
     
     
         23 . The method of  claim 14 , wherein the individual is a human. 
     
     
         24 . The method of  claim 14 , wherein the neural cell is a neural cell in the central nervous system. 
     
     
         25 . The method of  claim 14 , wherein the neural cell is a neural cell in the peripheral nervous system. 
     
     
         26 .- 32 . (canceled) 
     
     
         33 . The method of  claim 14 , wherein X is Er. 
     
     
         34 . The method of  claim 14 , wherein X is Tm. 
     
     
         35 . The method of  claim 14 , wherein X is Er/Tm. 
     
     
         36 . The method of  claim 14 , wherein the neural cell is a neural cell in the nucleus accumbens of the individual. 
     
     
         37 . The method of  claim 14 , wherein the light-responsive opsin comprises an amino acid sequence having at least 95% amino acid sequence identity to SEQ ID NO:2 or SEQ ID NO:3. 
     
     
         38 . The method of  claim 14 , wherein the light-responsive opsin comprises a NpHR protein. 
     
     
         39 . A method to hyperpolarize the plasma membrane of a neural cell in an individual comprising:
 (a) placing a plurality of lanthanide-doped nanoparticles in proximity to the neural cell, wherein the nanoparticles comprise NaYF 4 :Yb/X/Gd, wherein X is erbium (Er), thulium (Tm), or Er/Tm; and   (b) exposing the plurality of nanoparticles to electromagnetic radiation in the infrared (IR) or near infrared (NIR) spectrum, wherein the electromagnetic radiation in the IR or NIR spectrum is upconverted into green or blue light in the visible spectrum by the nanoparticles, and wherein a light-responsive opsin comprising a light-responsive proton pump is expressed on the plasma membrane and activation of the opsin by the light in the visible spectrum induces the hyperpolarization of the plasma membrane.   
     
     
         40 . A method to hyperpolarize the plasma membrane of a neural cell in an individual comprising:
 (a) administering a polynucleotide encoding a light-responsive opsin to an individual, wherein the light-responsive opsin comprises a light-responsive proton pump and is expressed on the plasma membrane of a neural cell in the individual and the opsin is capable of inducing membrane hyperpolarization of the neural cell when illuminated with light;   (b) administering a plurality of lanthanide-doped nanoparticles in proximity to the neural cell, wherein the nanoparticles comprise NaYF 4 :Yb/X/Gd, wherein X is erbium (Er), thulium (Tm), or Er/Tm; and   (c) exposing the plurality of nanoparticles to electromagnetic radiation in the infrared (IR) or near infrared (NIR) spectrum, wherein the electromagnetic radiation in the IR or NIR spectrum is upconverted into green or blue light in the visible spectrum and the activation of the opsin by the light in the visible spectrum induces the hyperpolarization of the plasma membrane.   
     
     
         41 . The method of  claim 39 , wherein the light-responsive opsin comprises an amino acid sequence having at least 85% amino acid sequence identity to SEQ ID NO:4. 
     
     
         42 . The method of  claim 39 , wherein the electromagnetic energy in the IR or NIR spectrum is upconverted into light having a wavelength of about 450 nm to about 495 nm. 
     
     
         43 . The method of  claim 39 , wherein the electromagnetic energy in the IR or NIR spectrum is upconverted into light having a wavelength corresponding to green light. 
     
     
         44 . The method of  claim 39 , wherein the electromagnetic energy in the IR or NIR spectrum is upconverted into light having a wavelength corresponding to blue light. 
     
     
         45 . The method of  claim 39 , wherein the individual is a non-human animal. 
     
     
         46 . The method of  claim 39 , wherein the individual is a human. 
     
     
         47 . The method of  claim 39 , wherein the neural cell is a neural cell in the central nervous system. 
     
     
         48 . The method of  claim 39 , wherein the neural cell is a neural cell in the peripheral nervous system. 
     
     
         49 . The method of  claim 39 , wherein X is Er. 
     
     
         50 . The method of  claim 39 , wherein X is Tm. 
     
     
         51 . The method of  claim 39 , wherein X is Er/Tm. 
     
     
         52 . The method of  claim 39 , wherein the light-responsive opsin comprises an amino acid sequence having at least 95% amino acid sequence identity to SEQ ID NO:4. 
     
     
         53 . The method of  claim 14 , wherein the light-responsive opsin comprises a GtR3 protein.