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M. Javaheri, C. Spee, K. Savalia, A. Ray, J.D. Weiland, M.S. Humayun, D.R. Hinton; The Effect of Electrical Stimulation on Retinal Muller Cells . Invest. Ophthalmol. Vis. Sci. 2006;47(13):5504.
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© ARVO (1962-2015); The Authors (2016-present)
The intraocular retinal prosthesis (IRP), when implanted at the epiretinal interface, rests on the internal limiting membrane (ILM). The ILM is primarily composed of Muller cell endfeet. During acute and chronic stimulation of the IRP, questions still exist about the exact interaction that takes place at the epiretinal–implant interface. More specifically, the long term effects of electrical stimulation on Muller cells have yet to be clearly defined.
A SV40 transformed, immortalized rat Muller cell line (kindly provided by Vijay Sarthy PhD) plated in a confluent monolayer was exposed to a concentric bipolar electrode with a flat tip configuration with an inner pole diameter of 75 um and an outer pole diameter of 300 um immersed in media, approximately 500um above the monolayer. A biphasic, balanced current pulse with 0.2 micro coulombs of charge per phase of pulse, with a pulse duration of 1millisecond was applied for one hour. Immunohistochemical studies were carried out with glial fibrillary acidic protein (GFAP)–specific, Ki67–specific, and ZO–1–specific antibodies.
Electrical stimulation of Muller cells demonstrated a marked increase in cell density and size at the site of electrical stimulation. Stimulated cells exhibited increased staining with GFAP, a marker for reactive gliosis in Muller cells, and Ki–67, a marker for proliferation in Muller cells. Stimulated cells showed equal levels of ZO–1, a marker for tight junctions in Muller cells, when compared to control.
We have developed and in–vitro model for Muller cells in order to establish conditions for a confluent monolayer of cells and a preliminary in–vitro model for electrical stimulation of Muller cells using a novel stimulating device that can apply a quantified amount of current to a specific area. Our preliminary studies have shown that a quantified amount of current, delivered to a specific area of Muller cells, can cause the cells to proliferate in that specific area, and upregulate GFAP and Ki–67.
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