Inhibition by thiocyanate of muscarinic-induced cytosolic acidification and Ca2%2b entry in rat sublingual acini
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Thiocyanate (SCN-) plays a critical part in an oral antimicrobial system by acting as a substrate for peroxidases. Salivary glands concentrate SCN- from blood up to 5 mM in saliva; however, the influence of SCN- on salivary acinar-cell function is unknown. The present study examined the effects of SCN- on the regulation of cytosolic pH (pHi) and free Ca2%2b concentration ([Ca2%2b]i) in rat sublingual mucous acini using the pH- and Ca2%2b-sensitive fluorescent indicators, 2′,7′-bis-(2-carboxyethyl)-5,6-carboxyfluorescein and fura-2, respectively. SCN- induced a concentration-dependent inhibition of the carbachol-stimulated cytosolic acidification (K1 2, approx. 1.4mM SCN-). Cytosolic pH recovery from an acid load was not changed by substitution of Cl- by SCN-, suggesting that Na%2b/H%2b exchange activity was not affected by SCN-. SCN- did not alter the initial carbachol-stimulated increase in [Ca2%2b]i; however, the sustained [Ca2%2b]i increase was inhibited by >65%25 (K1 2, approx. 1.0 mM SCN-). Furthermore, SCN- prevented the carbachol-stimulated Mn2%2b influx, indicating that it inhibits the divalent-cation entry pathway. Consistent with decreased Ca2%2b mobilization being involved in the blockade of the agonist-induced acidification by SCN-, only total replacement of Cl- with SCN- significantly inhibited the acidification induced by the Ca2%2b ionophore ionomycin. The permeability to SCN- through the Ca2%2b-dependent Cl- channels was 5.2-fold higher than the permeability to Cl-. These results suggest that inhibition of the agonist-induced cytosolic acidification by high-concentration SCN- may be mediated by both competitive inhibition of HCO3- efflux and by blockade of Ca2%2b influx. © 1995.
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anion channel; Ca2+ entry; cytosolic pH; salivary gland; SCN- 2',7' bis(carboxyethyl) 5(6) carboxyfluorescein; 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein; acid; bicarbonate; calcium; carbachol; chloride; diagnostic agent; fluorescein derivative; fluorescent dye; fura 2; ionomycin; manganese; muscarine; thiocyanate; thiocyanic acid derivative; animal; article; calcium channel; cell membrane potential; cytosol; drug effect; male; metabolism; pH; rat; rat strain; sublingual gland; Acids; Animal; Bicarbonates; Calcium; Calcium Channels; Carbachol; Chlorides; Cytosol; Fluoresceins; Fluorescent Dyes; Fura-2; Hydrogen-Ion Concentration; Ionomycin; Male; Manganese; Membrane Potentials; Muscarine; Rats; Rats, Wistar; Sublingual Gland; Support, Non-U.S. Gov't; Support, U.S. Gov't, P.H.S.; Thiocyanates
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