Soluble Adenylyl Cyclase Is Localized to Cilia and Contributes to Ciliary Beat Frequency Regulation via Production of cAMP
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 epithe...
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The Rockefeller University Press
2007
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| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2154360/ |
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pubmed-21543602008-01-17 Soluble Adenylyl Cyclase Is Localized to Cilia and Contributes to Ciliary Beat Frequency Regulation via Production of cAMP Schmid, Andreas Sutto, Zoltan Nlend, Marie-Christine Horvath, Gabor Schmid, Nathalie Buck, Jochen Levin, Lonny R. Conner, Gregory E. Fregien, Nevis Salathe, Matthias Articles 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. The Rockefeller University Press 2007-07 /pmc/articles/PMC2154360/ /pubmed/17591988 http://dx.doi.org/10.1085/jgp.200709784 Text en Copyright © 2007, The Rockefeller University Press This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). |
| repository_type |
Open Access Journal |
| institution_category |
Foreign Institution |
| institution |
US National Center for Biotechnology Information |
| building |
NCBI PubMed |
| collection |
Online Access |
| language |
English |
| format |
Online |
| author |
Schmid, Andreas Sutto, Zoltan Nlend, Marie-Christine Horvath, Gabor Schmid, Nathalie Buck, Jochen Levin, Lonny R. Conner, Gregory E. Fregien, Nevis Salathe, Matthias |
| spellingShingle |
Schmid, Andreas Sutto, Zoltan Nlend, Marie-Christine Horvath, Gabor Schmid, Nathalie Buck, Jochen Levin, Lonny R. Conner, Gregory E. Fregien, Nevis Salathe, Matthias Soluble Adenylyl Cyclase Is Localized to Cilia and Contributes to Ciliary Beat Frequency Regulation via Production of cAMP |
| author_facet |
Schmid, Andreas Sutto, Zoltan Nlend, Marie-Christine Horvath, Gabor Schmid, Nathalie Buck, Jochen Levin, Lonny R. Conner, Gregory E. Fregien, Nevis Salathe, Matthias |
| author_sort |
Schmid, Andreas |
| title |
Soluble Adenylyl Cyclase Is Localized to Cilia and Contributes to Ciliary Beat Frequency Regulation via Production of cAMP |
| title_short |
Soluble Adenylyl Cyclase Is Localized to Cilia and Contributes to Ciliary Beat Frequency Regulation via Production of cAMP |
| title_full |
Soluble Adenylyl Cyclase Is Localized to Cilia and Contributes to Ciliary Beat Frequency Regulation via Production of cAMP |
| title_fullStr |
Soluble Adenylyl Cyclase Is Localized to Cilia and Contributes to Ciliary Beat Frequency Regulation via Production of cAMP |
| title_full_unstemmed |
Soluble Adenylyl Cyclase Is Localized to Cilia and Contributes to Ciliary Beat Frequency Regulation via Production of cAMP |
| title_sort |
soluble adenylyl cyclase is localized to cilia and contributes to ciliary beat frequency regulation via production of camp |
| description |
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. |
| publisher |
The Rockefeller University Press |
| publishDate |
2007 |
| url |
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2154360/ |
| _version_ |
1611424270876934144 |