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Miscellaneous (18)

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Abstract #26881 Published in IGR 12-3

Focused Conference Group: P11 - G proteincoupled 7TM receptors: From molecular to physiological function rapid characterization of bimatoprost pharmacology in human ocular cells by using cellular dielectric specroscopy

Woodward D; Wang J; Piwnica D; Carling R; Cornell C; Martos J; Stamer D
Basic and Clinical Pharmacology and Toxicology 2010; 107: 659

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Bimatoprost is widely used as anti-glaucoma therapy but, to date, the cellular localization and characterization of its effects in individual ocular cell types has not been achieved. To accomplish this, primary cultures of human endothelial cells of Schlemm's canal (ECSC), trabecular meshwork (TM), and ciliary muscle (CM) cells were prepared from donor eyes. Cellular dielectric spectroscopy permits rapid pharmacological and second messenger characterization in primary cells in a 96 well format. It is a real time, non-invasive, label-free assay, which measures impedance. Bimatoprost produced a concentration-dependent change in cell monolayer impedance in ECSC, TM, and CM cells, with EC50 (nM) values of 1.6, 4.3, and 1.4, respectively. These effects were susceptible to the prostamide antagonist AGN 211334. Bimatoprost effects were insensitive to cholera toxin and pertussis toxin but were abolished by phorbol 12-myristate, 13-acetate. AGN 211334 exerted properties consistent with inverse agonism in TM cells but not ECSC cells . The pharmacology of bimatoprost and AGN 211334 in TM cells identified by cell dielectric spectroscopy was confirmed in its entirety by studying hydraulic conductivity in a TM cell monolayer. In conclusion a bimatoprost-sensitive prostamide receptor was identified and rapidly characterized using cellular dielectric spectroscopy. Bimatoprost potently stimulates ECSC, TM, and CM cells and the receptor is Gq coupled. Cellular dielectric spectroscopy represents a useful technique for rapid characterization of pharmacology and second messenger signaling in human primary cells.

D. Woodward. Allergan, Inc., Department of Biological Sciences, IrvineUnited States.

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Classification:

11.4 Prostaglandins (Part of: 11 Medical treatment)
3.6 Cellular biology (Part of: 3 Laboratory methods)

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