The role of K+ conductances in regulating membrane excitability in human gastric corpus smooth muscle

Changes in resting membrane potential (RMP) regulate membrane excitability. K+ conductance(s) are one of the main factors in regulating RMP. The functional role of K+ conductances has not been studied the in human gastric corpus smooth muscles (HGCS). To examine the role of K+ channels in regulation of RMP in HGCS we employed microelectrode recordings, patch-clamp, and molecular approaches. Tetraethylammonium and charybdotoxin did not affect the RMP, suggesting that BK channels are not involved in regulating RMP. Apamin, a selective small conductance Ca2+-activated K+ channel (SK) blocker, did not show a significant effect on the membrane excitability. 4-Aminopyridine, a Kv channel blocker, caused depolarization and increased the duration of slow wave potentials. 4-Aminopyridine also inhibited a delayed rectifying K+ current in isolated smooth muscle cells. End-product RT-PCR gel detected Kv1.2 and Kv1.5 in human gastric corpus muscles. Glibenclamide, an ATP-sensitive K+ channel (KATP) blocker, did not induce depolarization, but nicorandil, a KATP opener, hyperpolarized HGCS, suggesting that KATP are expressed but not basally activated. Kir6.2 transcript, a pore-forming subunit of KATP was expressed in HGCS. A low concentration of Ba2+, a Kir blocker, induced strong depolarization. Interestingly, Ba2+-sensitive currents were minimally expressed in isolated smooth muscle cells under whole-cell patch configuration. KCNJ2 (Kir2.1) transcript was expressed in HGCS. Unique K+ conductances regulate the RMP in HGCS. Delayed and inwardly rectifying K+ channels are the main candidates in regulating membrane excitability in HGCS. With the development of cell dispersion techniques of interstitial cells, the cell-specific functional significance will require further analysis.

Simultaneous monitoring of cellular depolarization and contraction: a new method to investigate excitability and contractility in isolated smooth muscle cells

The present experiments were performed to establish a method for simultaneous monitoring of excitation and contraction in isolated smooth muscle cells. The smooth muscle cells were dissociated from the colons of Wistar rats by enzymatic digestion. All the experiments were performed on mixtures of circular and longitudinal cells. In a first set of experiments, focal extracellular potentials (FEPs) and transmembrane action potentials (APs) were simultaneously recorded from the cells by use of extracellular and intracellular pipettes, respectively. In a second set of experiments, cellular contraction induced by chemical stimulation was monitored simultaneous with the FEP recordings. The FEPs had spike and plateau amplitudes of 44.5 ± 2.3 and 8.9 ± 0.7 mV, respectively, and reproduced the general morphology of gastrointestinal APs. The parallel mechanical measurements from the rat colonic cells showed that they shortened with an average peak contraction of 8.8 ± 1.4 μm and an average contraction velocity of 8.2 ± 0.9 μm/s, to develop an average peak force of 1.2 ± 0.2 μN, and generated an average peak power of 36 ± 15 pW. Simultaneous monitoring of FEPs and cellular contraction demonstrates correlations between the electrical and mechanical events taking place in the investigated cells.