Soluble adenylyl cyclase is localized to cilia and contributes to ciliary beat frequency regulation via production of cAMP

Andreas Schmid, Zoltan Sutto, Marie Christine Nlend, Gabor Horvath, Nathalie Schmid, Jochen Buck, Lonny R. Levin, Gregory E. Conner, Nevis Fregien, Matthias Salathe

Research output: Contribution to journalArticle

75 Scopus citations

Abstract

Ciliated airway epithelial cells are subject to sustained changes in intracellular CO2/HCO3- during exacerbations of airway diseases, but the role of CO2/HCO3- -sensitive soluble adenylyl cyclase (sAC) in ciliary beat regulation is unknown. We now show not only sAC expression in human airway epithelia (by RT-PCR, Western blotting, and immunofluorescence) but also its specific localization to the axoneme (Western blotting and immunofluorescence). Real time estimations of [cAMP] changes in ciliated cells, using FRET between fluorescently tagged PKA subunits (expressed under the foxj1 promoter solely in ciliated cells), revealed CO2/HCO3--mediated cAMP production. This cAMP production was specifically blocked by sAC inhibitors but not by transmembrane adenylyl cyclase (tmAC) inhibitors. In addition, this cAMP production stimulated ciliary beat frequency (CBF) independently of intracellular pH because PKA and sAC inhibitors were uniquely able to block CO2/HCO3 --mediated changes in CBF (while tmAC inhibitors had no effect). Thus, sAC is localized to motile airway cilia and it contributes to the regulation of human airway CBF. In addition, CO2/HCO3 - increases indeed reversibly stimulate intracellular cAMP production by sAC in intact cells.

Original languageEnglish (US)
Pages (from-to)99-109
Number of pages11
JournalJournal of General Physiology
Volume130
Issue number1
DOIs
StatePublished - Jul 2007

ASJC Scopus subject areas

  • Physiology

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