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Functional studies in the human submandibular duct cell line, HSG-PA, suggest a second salivary gland receptor subtype for nucleotides.

Extracellular nucleotides, acting through P2 purinoceptors, have been implicated in the regulation of ion transporting epithelia, including salivary gland acini. Multiple P2 purinoceptor subtypes have been suggested, including P2X, P2Y and P2U (or nucleotide) subtypes, as well as the P2Z subtype found in rat parotid acinar cells. We investigated responses to ATP, ATP analogs and UTP in transformed human submandibular gland duct cells (HSG-PA), in order to compare duct cell purinoreceptors with those in acinar cells. ATP, UTP and some ATP analogs increased, with different potencies, inositol phosphate accumulation, calcium mobilization and potassium efflux. Nucleotide-stimulated calcium mobilization occurred in the absence of, but was enhanced by, extracellular calcium, and maximal potassium efflux required extracellular calcium. UTP and ATP demonstrated equal potencies of about 1 microM and similar efficacies in eliciting these responses, and identical rank orders of potency for stimulating calcium mobilization and potassium efflux were obtained: UTP = ATP greater than ATP gamma S greater than ADP greater than ADP beta S, with alpha,beta-methylene-ATP and 2-methylthio-ATP having little or no effect. Agents reported to block nucleotide effects in parotid acini were ineffective in HSG-PA cells, and experiments in Mg(++)- and Ca(++)-free medium did not indicate that a form of ATP other than MgATP was the active species at the HSG-PA purinoceptor. The extracellular nucleotide effects were not altered by pertussis toxin. These results indicate the presence of a P2U or nucleotide receptor subtype in HSG-PA submandibular duct cells distinguishable from the P2Z purinoceptor of rat parotid acinar cells, suggesting involvement of multiple nucleotide receptor subtypes in salivary gland regulation.