Expression of store-operated Ca 2+ entry and transient receptor potential canonical and vanilloid-related proteins in rat distal pulmonary venous smooth muscle

Gongyong Peng, Wenju Lu, Xiaoyan Li, Yuqin Chen, Nanshan Zhong, Pixin Ran, Jian Wang

Research output: Contribution to journalArticlepeer-review

Abstract

Chronic hypoxia causes remodeling and alters contractile responses in both pulmonary arteries and pulmonary veins. Although pulmonary arteries have been studied extensively in these disorders, the mechanisms by which pulmonary veins respond to hypoxia and whether these responses contribute to chronic hypoxic pulmonary hypertension remain poorly understood. In pulmonary arterial smooth muscle, we have previously demonstrated that influx of Ca 2+ through store-operated calcium channels (SOCC) thought to be composed of transient receptor potential (TRP) proteins is likely to play an important role in development of chronic hypoxic pulmonary hypertension. To determine whether this mechanism could also be operative in pulmonary venous smooth muscle, we measured intracellular Ca 2+ concentration ([Ca 2+] i) by fura-2 fluorescence microscopy in primary cultures of pulmonary venous smooth muscle cells (PVSMC) isolated from rat distal pulmonary veins. In cells perfused with Ca 2+-free media containing cyclopiazonic acid (10 μM) and nifedipine (5 μM) to deplete sarcoplasmic reticulum Ca 2+ stores and block voltage-dependent Ca 2+ channels, restoration of extracellular Ca 2+ (2.5 mM) caused marked increases in [Ca 2+] i, whereas MnCl 2 (200 μM) quenched fura-2 fluorescence, indicating store-operated Ca 2+ entry (SOCE). SKF-96365 and NiCl 2, antagonists of SOCC, blocked SOCE at concentrations that did not alter Ca 2+ responses to 60 mM KCl. Of the seven known canonical TRP (TRPC1-7) and six vanilloid-related TRP channels (TRPV1-6), real-time PCR revealed mRNA expression of TRPC1>TRPC6>TRPC4> TRPC2 ≈ TRPC5 > TRPC3, TRPV2 > TRPV4 > TRPV1 in distal PVSMC, and TRPC1 > TRPC6 > TRPC3 > TRPC4 ≈ TRPC5, TRPV2 ≈ TRPV4 > TRPV1 in rat distal pulmonary vein (PV) smooth muscle. Western blotting confirmed protein expression of TRPC1, TRPC6, TRPV2, and TRPV4 in both PVSMC and PV. Our results suggest that SOCE through Ca 2+ channels composed of TRP proteins may contribute to Ca 2+ signaling in rat distal PV smooth muscle.

Original languageEnglish (US)
Pages (from-to)L621-L630
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Volume299
Issue number5
DOIs
StatePublished - Nov 2010

Keywords

  • Calcium signaling
  • Canonical transient receptor potential
  • Intracellular calcium concentration

ASJC Scopus subject areas

  • Physiology
  • Pulmonary and Respiratory Medicine
  • Physiology (medical)
  • Cell Biology

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