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Q. V. Hoang, H. Ripps; Gap Junctional Channel-Mediated Communication Between Retinal Pigment Epithelial Cells. Invest. Ophthalmol. Vis. Sci. 2008;49(13):5382.
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Gap junctional channels are found throughout the body, where they play a key role in signal transmission by allowing for direct cell-cell communication. Specifically, connexins mediating gap-junctional intercellular communication (GJIC) between RPE cells have been identified: Cx43 and Cx46. RPE GJIC has been previously studied by employing freeze-fracture, electron microscopy, RT-PCR and tracer techniques, but not electrophysiological recordings. Therefore, the present line of work attempts to develop a primary rat RPE cell culture suitable for electrophysiological experiments, allowing for the demonstration and characterization RPE GJIC.
Rat RPE cells were cultured from neonatal and adult rats, under various culture conditions, to develop an optimized method to study the GJIC between RPE cell pairs.
Rat eyecups with exposed RPE layers were trypsinized, and RPE cells were gently removed, dissociated in supplemented DMEM media and cultured for 4 to 14 days. Dual voltage-clamp electrophysiology experiments were performed, which demonstrated GJIC between RPE cell pairs among confluent dishes as well as isolated pairs.
A method of primary rat RPE cell culture was established, allowing for the electrophysiological demonstration of GJIC between RPE cells. Such a culture will allow for the characterization of RPE GJIC under normal and pathologic conditions.
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