Calcium-mediated, purinergic stimulation and polarized localization of calcium-sensitive adenylyl cyclase isoforms in human airway epithelia.
Purinergic stimulation of human airway epithelia results in a prolonged increase in ciliary beat frequency that depends on calcium-mediated cAMP production [Lieb, T., Wijkstrom Frei, C., Frohock, J.I., Bookman, R.J. and Salathe, M. (2002) Prolonged increase in ciliary beat frequency after short-term purinergic stimulation in human airway epithelial cells. J. Physiol. (Lond.) 538, 633-646]. Here, fully differentiated human airway epithelial cells in culture are shown to express calcium-stimulated transmembrane adenylyl cyclase (tmAC) isoforms (types 1, 3, and 8) by reverse transcription polymerase chain reaction. Immunohistochemistry of tracheal sections and fully differentiated airway epithelial cell cultures revealed polarized expression of these tmACs, with types 1 and 8 localized to the apical membrane and thus at the position required for ciliary regulation. Real-time, ciliated-cell specific cAMP production by tmACs upon apical, purinergic stimulation with UTP was confirmed using fluorescent energy resonance transfer between fluorescently tagged PKA subunits.
Nlend
Marie-Christine
MC
Division of Pulmonary and Critical Care Medicine, University of Miami Miller School of Medicine, Miami, FL, USA.
Schmid
Andreas
A
Sutto
Zoltan
Z
Ransford
George A
GA
Conner
Gregory E
GE
Fregien
Nevis
N
Salathe
Matthias
M
D000262:Adenylyl Cyclases / Q000235:genetics / Q000378:metabolism*; D002118:Calcium / Q000494:pharmacology*; D016764:Cell Polarity / Q000502:physiology*; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000378:metabolism; D004847:Epithelial Cells / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006801:Humans; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D011687:Purines / Q000494:pharmacology*; D020545:Respiratory Mucosa / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014018:Tissue Distribution
D007527:Isoenzymes; D011687:Purines; D000242:Cyclic AMP; D000262:Adenylyl Cyclases; D002118:Calcium