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1 Division of Pulmonary and Critical Care Medicine, Johns Hopkins School of Medicine, Baltimore, Maryland, USA
2 Division of Pulmonary and Critical Care Medicine, Johns Hopkins School of Medicine, Baltimore, Maryland, USA; Department of Physiology and Pathophysiology, Fujian Medical University, Fuzhou, Fujian, China
* To whom correspondence should be addressed. E-mail: jsks{at}welchlink.welch.jhu.edu.
Transient receptor potential melastatin- (TRPM) and vanilloid-related (TRPV) channels are non-selective cation channels pertinent to diverse physiological functions. Multiple TRPM and TRPV channel subtypes have been identified and cloned in different tissues. However, their information in vascular tissue is scant. In this study, we sought to identify TRPM and TRPV channel subtypes expressed in rat de-endothelialized intralobar pulmonary arteries (PAs) and aorta. Using RT-PCR, mRNA of TRPM2, TRPM3, TRPM4, TRPM7 and TRPM8 of TRPM family and TRPV1, TRPV2, TRPV3 and TRPV4 of TRPV family were detected in both PAs and aorta. Quantitative real-time RT-PCR showed that TRPM8 and TRPV4 were the most abundantly expressed TRPM and TRPV subtypes, respectively. Moreover, western blot analysis verified the expression of TRPM2, TRPM8, TRPV1, and TRPV4 proteins in both types of vascular tissue. To examine the functional activities of these channels, [Ca2+]i transients were monitored in pulmonary arterial smooth muscle cells (PASMCs) and aortic smooth muscle cells (ASMCs). The TRPM8 agonist menthol (300 µM), and the TRPV4 agonist 4
-phorbol 12,13-didecanoate (1 µM) evoked significant increases in [Ca2+]i in PASMCs and ASMCs. These Ca2+ responses were obliterated in the absence of extracellular Ca2+ or in the presence of 300 µM Ni2+, but were unaffected by 1 µM nifedipine, suggesting Ca2+ influx via non-selective cation channels. Hence, for the first time, our results indicate that multiple functional TRPM and TRPV channels are co-expressed in rat intralobar PAs and aorta. These novel Ca2+ entry pathways may play important roles in the regulation of pulmonary and systemic circulation.
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