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Am J Physiol Lung Cell Mol Physiol 281: L832-L843, 2001;
1040-0605/01 $5.00
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Vol. 281, Issue 4, L832-L843, October 2001

Human airway smooth muscle expresses 7 isoforms of adenylyl cyclase: a dominant role for isoform V

Dingbang Xu1, Cary Isaacs1, Ian P. Hall2, and Charles W. Emala1

1 Department of Anesthesiology, College of Physicians and Surgeons of Columbia University, New York, New York 10032; and 2 Department of Therapeutics, Institute of Cell Signaling, University Hospital of Nottingham, Nottingham NG7 2UH, United Kingdom

Adenylyl cyclases are a nine-member family of differentially regulated enzymes responsible for the synthesis of cAMP. cAMP is an important second messenger that contributes to the regulation of airway smooth muscle tone. However, little is known regarding the expression and regulation of adenylyl cyclase isoforms in airway smooth muscle cells. Nondegenerate specific primers were designed for all nine known isoforms of human adenylyl cyclase. RT-PCR experiments were performed using total RNA extracted from whole human brain (positive control), whole rat brain (negative control), whole human trachea, human airway smooth muscle, and primary cultures of human airway smooth muscle cells. Seven of the nine known isoforms of adenylyl cyclase (isoforms I, III-VII, and IX) were expressed at the mRNA level in both human airway smooth muscle and primary cultures of human airway smooth muscle cells. Immunoblot and adenylyl cyclase functional assay indicated that isoform V is likely among the functionally predominant isoforms of adenylyl cyclase in human airway smooth muscle. These results suggest that multiple isoforms of adenylyl cyclase enzymes are coexpressed in human airway smooth muscle cells and that isoform V is among the functionally important isoforms.

reverse transcriptase-polymerase chain reaction; trachea; adenosine 3',5'-cyclic monophosphate; immunoblot; Sp-diasteromer of adenosine 3',5'-cyclic monophosphothioate; protein kinase A; protein kinase C


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