Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents

This Article Full Text Full Text (PDF, 2176K) PPT slides of all figures Alert me when this article is cited Citation Map Services Email this article Similar articles in this journal Similar articles in PubMed Alert me to new content in the JGP Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Schmid, A. Articles by Salathe, M. Search for Related Content PubMed PubMed Citation Articles by Schmid, A. Articles by Salathe, M. Social Bookmarking                      What's this?

¹ Division of Pulmonary and Critical Care and ² Department of Cell Biology and Anatomy, University of Miami, Miami, FL 33136
³ Department of Respiratory Medicine, Semmelweis University, Budapest, Hungary
⁴ Department of Pharmacology, Joan and Sanford I. Weill Medical College of Cornell University, New York, NY 10021

Correspondence to Matthias Salathe: msalathe{at}miami.edu

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.

Abbreviations used in this paper: ALI, air-liquid interface; CBF, ciliary beat frequency; FRET, fluorescence resonance energy transfer; ORF, open reading frame; sAC, soluble adenylyl cyclase; tmAC, transmembrane adenylyl cyclase.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				A. I. Masyuk, S. A. Gradilone, J. M. Banales, B. Q. Huang, T. V. Masyuk, S.-O. Lee, P. L. Splinter, A. J. Stroope, and N. F. LaRusso Cholangiocyte primary cilia are chemosensory organelles that detect biliary nucleotides via P2Y12 purinergic receptors Am J Physiol Gastrointest Liver Physiol, October 1, 2008; 295(4): G725 - G734. [Abstract] [Full Text] [PDF]

				L. S. Ramos, J. H. Zippin, M. Kamenetsky, J. Buck, and L. R. Levin Glucose and GLP-1 Stimulate cAMP Production via Distinct Adenylyl Cyclases in INS-1E Insulinoma Cells J. Gen. Physiol., August 25, 2008; 132(3): 329 - 338. [Abstract] [Full Text] [PDF]

				S. W. Moore, K. Lai Wing Sun, F. Xie, P. A. Barker, M. Conti, and T. E. Kennedy Soluble Adenylyl Cyclase Is Not Required for Axon Guidance to Netrin-1 J. Neurosci., April 9, 2008; 28(15): 3920 - 3924. [Abstract] [Full Text] [PDF]

Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents