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Lu Yin, Benjamin Masella, Deniz Dalkara, John.G. Flannery, David. V. Schaffer, David. R. Williams, William. H. Merigan; In vivo imaging of ganglion cell physiology in macaque fovea using a calcium indicator. Journal of Vision 2012;12(14):55. doi: https://doi.org/10.1167/12.14.55.
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© ARVO (1962-2015); The Authors (2016-present)
The physiological response of macaque retinal ganglion cells near the fovea was studied in vivo by imaging of their calcium response using a genetically encoded calcium indicator. G-CaMP5 was inserted into the ganglion cells by intravitreal injection of 7m8-CMV-G-CaMP5. 7m8 is a novel adeno-associated viral (AAV) variant created by directed evolution that shows promise to be the most effective vector at present for transducing macaque retina through intravitreal injection. The presence of G-CaMP5 was detected within 3 weeks of injection by low-resolution fluorescence imaging of the retina. High-resolution experiments were then performed using an adaptive optics scanning laser ophthalmoscope. Visual activation of retinal ganglion cells was accomplished by either flashed presentation of long-wavelength (peak 590 nm) light or modulation of a 488 nm laser. In the latter case, the spatial separation of the foveal cones and their associated ganglion cells allowed for simultaneous imaging and stimulation with a single laser source. Both methods produced robust activation of retinal ganglion cells. Individual ganglion cells showed a variety of temporal response profiles. Control experiments demonstrated that the fluorecence responses represented physiological activation. This method measures the physiology of individual macaque foveal ganglion cells in vivo. Physiological responses from these cells can be measured repeatedly over an extended period of time, which provides the possibility of studying the functional consequences of age, disease or injury in addition to improving our understanding of normal retinal function.
Meeting abstract presented at OSA Fall Vision 2012
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