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Published online before print April 11, 2007, 10.1183/09031936.00008507
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Eur Respir J 2007; 30:269-278
Copyright ©ERS Journals Ltd 2007

Store-refilling involves both L-type calcium channels and reverse-mode sodium–calcium exchange in airway smooth muscle

S. Hirota and L. J. Janssen

Firestone Institute for Respiratory Health, St. Joseph’s Healthcare and Dept of Medicine, McMaster University, Hamilton, ON, Canada.

CORRESPONDENCE: L. J. Janssen, L-314, St. Joseph’s Healthcare, 50 Charlton Ave. East, Hamilton, ON, Canada L8N 4A6. Fax: 1 9055406510. E-mail: janssenl{at}mcmaster.ca

Keywords: Airway smooth muscle, calcium-handling, L-type calcium channel, membrane depolarisation, reverse-mode sodium–calcium exchanger, sarcoplasmic reticulum

Received: January 22, 2007
Accepted March 29, 2007

The aim of the present study was to examine the relative contributions to store-refilling of the two following voltage-regulated calcium ion influx pathways: 1) L-type-Ca2+ channels; and 2) the reverse-mode of the sodium–calcium exchanger (NCX).

Successive acetylcholine-induced contractions, triggered in bovine tracheal smooth muscle strips, were measured to determine the effect of intervention on contractions as an indication of the extent of store-refilling.

Pre-treating the tissues with cromakalim significantly reduced the magnitude of successive contractions. Zero-Ca2+ bathing media abolished the contractions, an effect that was completely reversed upon reintroduction of Ca2+. Inhibition of L-type Ca2+ channels, with nifedipine, significantly reduced the magnitude of the contractions. Similarly, inhibition of the reverse-mode of the NCX, with KB-R7943, significantly reduced the magnitude of the contractions. However, neither nifedipine nor KB-R7943 alone reduced contractions to the same extent as observed under zero-Ca2+ conditions. Concurrent treatment with nifedipine and KB-R7943 almost abolished successive contractions. Furthermore, concurrent treatment with nifedipine and zero-Na+ bathing media displayed a significantly greater effect than nifedipine alone. Probing the expression of NCX1 isoforms by Western blotting revealed the presence of three bands of 160, 120 and 110 kDa. The 120- and 110-kDa bands were identified as variably spliced NCX isoforms, NCX1.1 and NCX1.3, respectively.

Taken together, these data suggest that influx of calcium ions through both L-type calcium channels and the reverse-mode of the sodium–calcium exchanger is necessary for complete store-refilling in airway smooth muscle.




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