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Screening of gap junction antagonists on dye coupling in the rabbit retina

Published online by Cambridge University Press:  22 August 2007

FENG PAN ,
STEPHEN L. MILLS  and
STEPHEN C. MASSEY
FENG PAN
Affiliation:
Department of Ophthalmology and Visual Science, University of Texas Medical School at Houston, Houston Texas
STEPHEN L. MILLS
Affiliation:
Department of Ophthalmology and Visual Science, University of Texas Medical School at Houston, Houston Texas
STEPHEN C. MASSEY
Affiliation:
Department of Ophthalmology and Visual Science, University of Texas Medical School at Houston, Houston Texas
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Abstract

Many cell types in the retina are coupled via gap junctions and so there is a pressing need for a potent and reversible gap junction antagonist. We screened a series of potential gap junction antagonists by evaluating their effects on dye coupling in the network of A-type horizontal cells. We evaluated the following compounds: meclofenamic acid (MFA), mefloquine, 2-aminoethyldiphenyl borate (2-APB), 18-α-glycyrrhetinic acid, 18-β-glycyrrhetinic acid (18-β-GA), retinoic acid, flufenamic acid, niflumic acid, and carbenoxolone. The efficacy of each drug was determined by measuring the diffusion coefficient for Neurobiotin (Mills & Massey, 1998). MFA, 18-β-GA, 2-APB and mefloquine were the most effective antagonists, completely eliminating A-type horizontal cell coupling at a concentration of 200 μM. Niflumic acid, flufenamic acid, and carbenoxolone were less potent. Additionally, carbenoxolone was difficult to wash out and also may be harmful, as the retina became opaque and swollen. MFA, 18-β-GA, 2-APB and mefloquine also blocked coupling in B-type horizontal cells and AII amacrine cells. Because these cell types express different connexins, this suggests that the antagonists were relatively non-selective across several different types of gap junction. It should be emphasized that MFA was water-soluble and its effects on dye coupling were easily reversible. In contrast, the other gap junction antagonists, except carbenoxolone, required DMSO to make stock solutions and were difficult to wash out of the preparation at the doses required to block coupling in A-type HCs. The combination of potency, water solubility and reversibility suggest that MFA may be a useful compound to manipulate gap junction coupling.

Keywords

Horizontal cellsRetinaGap junction antagonist
Type
Research Article
Information
Visual Neuroscience , Volume 24 , Issue 4 , July 2007 , pp. 609 - 618
Copyright
© 2007 Cambridge University Press

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