Modulation of chloride secretion across the pigmented rabbit conjunctiva
Abstract
The purpose of the present study was to investigate whether active Cl- secretion in the pigmented rabbit conjunctiva was subject to cAMP, Ca2+ and protein kinase C (PKC) modulation. The excised pigmented rabbit conjunctivas were mounted in the modified Ussing-type chambers for measurement of unidirectional 36Cl fluxes under the open-circuit condition and of the short-circuit current (Isc), potential difference, and transconjunctival electrical resistance. The results indicate that Cl- secretion across the conjunctiva was abolished by mucosal application of 1 mM N-phenylanthranilic acid and was reduced by 40% by serosal application of 10 microM bumetanide. Net Cl- flux was stimulated by 133% by 1 mM 8-Br cAMP, 107% by 10 microM A23187, and 87% by 1 microM phorbol 12-myristate-13-acetate (PMA), suggesting that cAMP, Ca2+, and PKC all modulated active Cl- secretion, respectively. There existed a linear correlation between measured changes in net Cl- flux and observed changes in Isc (r2=0.99). The serial treatment of the conjunctiva with (a) 1 mM 8-Br cAMP and 10 microM A23187 and (b) 10 microM A23187 and 1 microM PMA resulted in sequence-independent, additive stimulation of Isc. In the case of 1 mM 8-Br cAMP and 1 microM PMA, addi...Continue Reading
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