May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Gap Junctional Channel-Mediated Communication Between Retinal Pigment Epithelial Cells
Author Affiliations & Notes
  • Q. V. Hoang
    Ophthalmology & Visual Sciences, Univ of Illinois at Chicago, Chicago, Illinois
    Grass Lab, Marine Biological Laboratory, Woods Hole, Massachusetts
  • H. Ripps
    Ophthalmology & Visual Sciences, Univ of Illinois at Chicago, Chicago, Illinois
  • Footnotes
    Commercial Relationships  Q.V. Hoang, None; H. Ripps, None.
  • Footnotes
    Support  Grass Foundation Fellowship (QH), RPB Senior Scientific Investigator Award (HR) and NEI Core Grant EY-01792
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 5382. doi:
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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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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : 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.

Methods: : 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.

Results: : 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.

Conclusions: : 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.

Keywords: retinal pigment epithelium • gap junctions/coupling • electrophysiology: non-clinical 
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