VIP and muscarinic synergistic mucin secretion by salivary mucous cells is mediated by enhanced PKC activity via VIP-induced release of an intracellular Ca2+ pool

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Mucin secretion by salivary mucous glands is mediated predominantly by parasympathetic acetylcholine activation of cholinergic muscarinic receptors via increased intracellular free calcium ([Ca2+]i) and activation of conventional protein kinase C isozymes (cPKC). However, the parasympathetic co-neurotransmitter, vasoactive intestinal peptide (VIP), also initiates secretion, but to a lesser extent. In the present study, cross talk between VIP- and muscarinic-induced mucin secretion was investigated using isolated rat sublingual tubuloacini. VIP-induced secretion is mediated by cAMP-activated protein kinase A (PKA), independently of increased [Ca2+]i. Synergistic secretion between VIP and the muscarinic agonist, carbachol, was demonstrated but only with submaximal carbachol. Carbachol has no effect on cAMP ± VIP. Instead, PKA activated by VIP releases Ca2+ from an intracellular pool maintained by the sarco/endoplasmic reticulum Ca2+-ATPase pump. Calcium release was independent of phospholipase C activity. The resultant sustained [Ca2+]i increase is additive to submaximal, but not maximal carbachol-induced [Ca2+]i. Synergistic mucin secretion was mimicked by VIP plus either phorbol 12-myristate 13-acetate or 0.01 μM thapsigargin, and blocked by the PKC inhibitor, Gö6976. VIP-induced Ca2+ release also promoted store-operated Ca2+ entry. Synergism is therefore driven by VIP-mediated [Ca2+]i augmenting cPKC activity to enhance muscarinic mucin secretion. Additional data suggest ryanodine receptors control VIP/PKA-mediated Ca2+ release from a Ca2+ pool also responsive to maximal carbachol. A working model of muscarinic and VIP control of mucous cell exocrine secretion is presented. Results are discussed in relation to synergistic mechanisms in other secretory cells, and the physiological and therapeutic significance of VIP/muscarinic synergism controlling salivary mucous cell exocrine secretion.

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We thank Dr. M. Fallon and Ms. Linda A. Richardson for technical assistance, and Dr. J. Melvin for technical advice and discussions. This work was supported by National Institutes of Health / National Institute of Dental and Craniofacial Research grants DE10480 and DE014730 to D. Culp. The funders had no role in study design, data collection and analysis, decisions to publish, or manuscript preparation.

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D. Culp contributed to conception and experimental design of the study, data analysis, interpretation of results, and drafted and critically revised the manuscript; R. L. Evans contributed to the experimental design, data interpretation and critically revised the manuscript; Z. Zhang contributed to the experimental design, data acquisition, data interpretation and drafted and critically revised the manuscript. All authors gave final approval and agree to be accountable for all aspects of the work.

Correspondence to David J. Culp.

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Culp, D.J., Zhang, Z. & Evans, R.L. VIP and muscarinic synergistic mucin secretion by salivary mucous cells is mediated by enhanced PKC activity via VIP-induced release of an intracellular Ca2+ pool. Pflugers Arch - Eur J Physiol (2020) doi:10.1007/s00424-020-02348-7

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  • Salivary glands
  • Mucous cells
  • Mucins
  • Signal transductions
  • Saliva