In retinal disease, despite the loss of light sensitivity as photoreceptors die, many retinal interneurons survive in a physiologically and metabolically functional state for long periods. This provides an opportunity for treatment by genetically adding a light sensitive function to these cells. Optogenetic therapies are in development, but, to date, they have suffered from low light sensitivity and narrow dynamic response range of microbial opsins. Expression of light-sensitive G protein coupled receptors (GPCRs), such as vertebrate rhodopsin, can increase sensitivity by signal amplification, as shown by several groups. Here, we describe the methods to (1) express light gated GPCRs in retinal neurons, (2) record light responses in retinal explants in vitro, (3) record cortical light responses in vivo, and (4) test visually guided behavior in treated mice.
CITATION STYLE
Gaub, B. M., Berry, M. H., Visel, M., Holt, A., Isacoff, E. Y., & Flannery, J. G. (2018). Optogenetic retinal gene therapy with the light gated GPCR vertebrate rhodopsin. In Methods in Molecular Biology (Vol. 1715, pp. 177–189). Humana Press Inc. https://doi.org/10.1007/978-1-4939-7522-8_12
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