TY - JOUR
T1 - Calcium-mediated, purinergic stimulation and polarized localization of calcium-sensitive adenylyl cyclase isoforms in human airway epithelia
AU - Nlend, Marie Christine
AU - Schmid, Andreas
AU - Sutto, Zoltan
AU - Ransford, George A.
AU - Conner, Gregory E.
AU - Fregien, Nevis
AU - Salathe, Matthias
PY - 2007/7/10
Y1 - 2007/7/10
N2 - 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.
AB - 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.
KW - Adenylyl cyclase isoforms
KW - Airway epithelium
KW - cAMP production
KW - Ciliary beating
KW - Polarized expression
UR - http://www.scopus.com/inward/record.url?scp=34250876860&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=34250876860&partnerID=8YFLogxK
U2 - 10.1016/j.febslet.2007.06.015
DO - 10.1016/j.febslet.2007.06.015
M3 - Article
C2 - 17586501
AN - SCOPUS:34250876860
VL - 581
SP - 3241
EP - 3246
JO - FEBS Letters
JF - FEBS Letters
SN - 0014-5793
IS - 17
ER -