Constrictor-induced translocation of NFAT3 in human and rat pulmonary artery smooth muscle.
The transcription factor nuclear factor of activated T cells (NFAT) resides in the cytoplasm in resting cells and upon stimulation is dephosphorylated, translocates to the nucleus, and becomes transcriptionally active. NFAT is commonly activated by stimulation of receptors coupled to Ca(2+) mobilization; however, little is known about the regulation of NFAT in pulmonary vascular smooth muscle. The aim of this study was to investigate regulation of NFAT in human and rat intralobar pulmonary artery by two constrictors: phenylephrine (PE) and 20-hydroxyeicosatetraenoic acid (20-HETE), a cytochrome P-450 metabolite formed endogenously in lungs. Immunostaining of smooth muscle cells revealed cytoplasmic localization of NFAT in untreated cells, and PE or 20-HETE induced translocation to the nucleus, with maximal effect at 30 min. Cyclosporin A and FK-506 (both 1 microM) inhibited NFAT translocation, indicating involvement of calcineurin. Moreover, the Rho-kinase blocker Y-27632 prevented translocation. Translocation of NFAT was confirmed by Western blots, with NFAT3 the prominent isoform in pulmonary artery. Constrictors caused calcineurin-sensitive translocation of NFAT to nuclei in intact arteries, demonstrating regulation in native tissue. To investigate a role for Ca(2+), cells were loaded with fura-2. Whereas PE caused an acute transient rise of [Ca(2+)](i), 20-HETE caused a prolonged low amplitude rise of [Ca(2+)](i). The involvement of Rho-kinase in PE- and 20-HETE-induced NFAT3 translocation in pulmonary artery suggests a level of control not previously recognized in smooth muscle. Constrictors of the pulmonary vasculature not only cause acute responses but also activate NFAT, which may alter gene expression in pulmonary health and disease.
Yaghi
Asma
A
Dept. of Physiology and Pharmacology, University of Western Ontario, London, Ontario, Canada N6A 5C1.
Sims
Stephen M
SM
D021581:Active Transport, Cell Nucleus / Q000187:drug effects / Q000502:physiology; D000577:Amides / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000378:metabolism; D020013:Calcium Signaling / Q000187:drug effects / Q000235:genetics; D002467:Cell Nucleus / Q000378:metabolism*; D016572:Cyclosporine / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects / Q000502:physiology; D006801:Humans; D006893:Hydroxyeicosatetraenoic Acids / Q000494:pharmacology*; D008171:Lung Diseases / Q000378:metabolism; D008297:Male; D009119:Muscle Contraction / Q000187:drug effects / Q000502:physiology*; D009130:Muscle, Smooth / Q000166:cytology / Q000378:metabolism*; D050778:NFATC Transcription Factors / Q000378:metabolism*; D009924:Organ Culture Techniques; D010656:Phenylephrine / Q000494:pharmacology*; D020033:Protein Isoforms / Q000378:metabolism; D011651:Pulmonary Artery / Q000166:cytology / Q000378:metabolism*; D011725:Pyridines / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D014662:Vasoconstrictor Agents / Q000494:pharmacology*
D000577:Amides; D004791:Enzyme Inhibitors; D006893:Hydroxyeicosatetraenoic Acids; D050778:NFATC Transcription Factors; C494010:NFATC4 protein, human; D020033:Protein Isoforms; D011725:Pyridines; D014662:Vasoconstrictor Agents; C108830:Y 27632; D010656:Phenylephrine; C055987:20-hydroxy-5,8,11,14-eicosatetraenoic acid; D016572:Cyclosporine; D002118:Calcium