RESUMO
The study explored the potential of an animal opsin nonselectively expressed in various neuronal elements of the degenerative retina to restore the impaired visual function. A knockout murine model of inherited retinal dystrophy was used. Mice were injected intravitreally with either a virus carrying the gene of short-wavelength cone opsin associated with a reporter fluorescent protein or a control virus carrying the sequence of a modified fluorescent protein with enhanced membrane tropism. Viral transduction induced pronounced opsin expression in ganglion, bipolar, and horizontal retinal neurons. Behavioral testing included the visually guided task in the trapezoid Morris water maze and showed a partial recovery of the learning ability in the mice whose retinas had been transduced with cone opsin.
Assuntos
Opsinas dos Cones , Degeneração Retiniana , Camundongos , Animais , Opsinas dos Cones/genética , Opsinas dos Cones/metabolismo , Degeneração Retiniana/genética , Degeneração Retiniana/metabolismo , Células Fotorreceptoras Retinianas Cones/metabolismo , Retina , Opsinas/metabolismo , Camundongos KnockoutRESUMO
To perform optogenetic prosthetics of the retinal ganglion cell receptive field, a bicistronic genetic construct carrying the genes encoding the excitatory (channelrhodopsin-2) and inhibitory (Guillardia theta anion channelrhodopsin GtACR2) rhodopsins was created. A characteristic feature of this construct was the combination of these two genes with a mutant IRES insertion between them, which ensures the exact ratio of expression levels of the first and second genes in each transfected cell. Illumination of the central part of the neuron with light with a wavelength of 470 nm induced the action potential generation in the cell. Stimulation of the peripheral neuronal region with light induced the inhibition of action potential generation. Thus, using optogenetics methods, we simulated the ON-OFF interaction in the retinal ganglion cell receptive field. Theoretically, this construct can be used for optogenetic prosthetics of degenerative retina in the case of its delivery to the ganglion cells with lentiviral vectors.
