Affiliation:
1. Smooth Muscle Research Centre, Dundalk Institute of Technology, Dundalk, Ireland
Abstract
A collagenase-proteinase mixture was used to isolate airway smooth muscle cells (ASMC) from rabbit bronchi, and membrane currents were recorded using the whole cell patch-clamp technique. Stepping from −100 mV to a test potential of −40 mV evoked a fast voltage-dependent Na+ current, sometimes with an amplitude of several nanoamperes. The current disappeared within 15 min of exposure to papain + DTT ( n = 6). Comparison of the current in ASMC with current mediated by NaV1.5 α-subunits expressed in human embryonic kidney cells revealed similar voltage dependences of activation ( V1/2 = −42 mV for NaV1.5) and sensitivities to TTX (IC50 = 1.1 and 1.2 μM for ASMC and NaV1.5, respectively). The current in ASMC was also blocked by lidocaine (IC50 = 160 μM). Although veratridine, an agonist of voltage-gated Na+ channels, reduced the peak current by 33%, it slowed inactivation, resulting in a fourfold increase in sustained current (measured at 25 ms after onset). In current-clamp mode, veratridine prolonged evoked action potentials from 37 ± 9 to 1,053 ± 410 ms ( n = 8). Primers for NaV1.2–1.9 were used to amplify mRNA from groups of ∼20 isolated ASMC and from whole bronchial tissue by RT-PCR. Transcripts for NaV1.2, NaV1.3, and NaV1.5–1.9 were detected in whole tissue, but only NaV1.2 and NaV1.5 were detected in single cells. We conclude that freshly dispersed rabbit ASMC express a fast voltage-gated Na+ current that is mediated mainly by the NaV1.5 subtype.
Publisher
American Physiological Society
Cited by
13 articles.
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