scholarly journals Control of ion distribution in isolated smooth muscle cells. I. Potassium.

1980 ◽  
Vol 75 (2) ◽  
pp. 163-182 ◽  
Author(s):  
C R Scheid ◽  
F S Fay
Keyword(s):  
Smooth Muscle ◽  
Membrane Potential ◽  
Smooth Muscle Cells ◽  
Muscle Cells ◽  
Resting Membrane Potential ◽  
Stomach Muscle ◽  
Isolated Cells ◽  
Intracellular Space ◽  
Ion Movements ◽  
Isolated Smooth Muscle Cells

We describe a technique for examining unidirectional ion movements in suspensions of enzymatically disaggregated smooth muscle cells derived from stomach muscle of the toad. This technique has been used to analyze the movement of 42K across these cells. This analysis was greatly simplified by the finding that the cells were in a steady state with respect to K+ distribution after isolation. The potassium contents of the isolated cells were identical to those of intact smooth muscle (131 mM/liter intracellular fluid) and stable for over 4 h; moreover, the unidirectional influx and efflux rates were equal. An additional simplification was provided by the finding that virtually all the K+ exchanges in a manner predicted for a simple two-compartment system consisting of an extracellular and an intracellular space. Transmembrane K+ flux in these cells averaged 1.2 pmol.cm-2.s-1 at room temperature. A large portion (approximately 80%) of 42K influx appeared to be mediated by a saturable transport system with an apparent Km of 0.6 mM and an apparent Vmax of 1.3 pmol.cm-2.s-1. The calculated resting membrane permeability to K+ in these isolated smooth muscle cells, assuming a membrane potential of -50 mV, was 2.9 X 10(-8) cm/s. The calculated gK+ was 2.7 mumho/cm2 constituting only a small fraction of the total membrane conductance as measured electrophysiologically. The latter finding suggests that the resting membrane potential in the isolated cells must be determined by ions in addition to K+. We propose that these methods for studying ion movements in smooth muscle should aid in unraveling the mechanisms responsible for controlling the distribution of ions both at rest, as in the present study, as well as in response to neurotransmitters.

Role of HERG-like K+ currents in opossum esophageal circular smooth muscle

1999 ◽  
Vol 277 (6) ◽  
pp. C1284-C1290 ◽  
Author(s):  
Hamid I. Akbarali ◽  
Hemant Thatte ◽  
Xue Dao He ◽  
Wayne R. Giles ◽  
Raj K. Goyal
Keyword(s):  
Smooth Muscle ◽  
Membrane Potential ◽  
Smooth Muscle Cells ◽  
Single Cells ◽  
Circular Muscle ◽  
Muscle Cells ◽  
Resting Membrane Potential ◽  
Channel Blockers ◽  
Circular Smooth Muscle ◽  
Inward Currents

An inwardly rectifying K+ conductance closely resembling the human ether-a-go-go-related gene (HERG) current was identified in single smooth muscle cells of opossum esophageal circular muscle. When cells were voltage clamped at 0 mV, in isotonic K+ solution (140 mM), step hyperpolarizations to −120 mV in 10-mV increments resulted in large inward currents that activated rapidly and then declined slowly (inactivated) during the test pulse in a time- and voltage- dependent fashion. The HERG K+ channel blockers E-4031 (1 μM), cisapride (1 μM), and La3+ (100 μM) strongly inhibited these currents as did millimolar concentrations of Ba2+. Immunoflourescence staining with anti-HERG antibody in single cells resulted in punctate staining at the sarcolemma. At membrane potentials near the resting membrane potential (−50 to −70 mV), this K+ conductance did not inactivate completely. In conventional microelectrode recordings, both E-4031 and cisapride depolarized tissue strips by 10 mV and also induced phasic contractions. In combination, these results provide direct experimental evidence for expression of HERG-like K+ currents in gastrointestinal smooth muscle cells and suggest that HERG plays an important role in modulating the resting membrane potential.

Dissociation of contraction and muscarinic receptor binding to isolated smooth muscle cells

1986 ◽  
Vol 251 (4) ◽  
pp. G546-G552 ◽  
Author(s):  
S. M. Collins ◽  
D. J. Crankshaw
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Binding Sites ◽  
Specific Binding ◽  
Muscle Cells ◽  
Muscarinic Agonists ◽  
Isolated Cells ◽  
Single Class ◽  
Antagonist Binding ◽  
Isolated Smooth Muscle Cells

We examined changes in [3H]QNB binding and cell length induced by muscarinic ligands in a suspension of single smooth muscle cells isolated from the canine stomach. Cells contracted following a brief (30 s) exposure to picomolar concentrations of muscarinic agonists and yielded ED50 values of 1.0 +/- 0.7 pM for oxotremorine, 12.5 +/- 1.8 pM for carbachol, and 16.0 +/- 2.9 pM for metacholine. Contraction was inhibited by atropine with a pA2 value of 10.2 +/- 1.1. The binding of [3H]QNB was rapid and reversible and was stereospecific and pharmacologically appropriate. Specific binding of [3H]QNB was saturable and bound with high affinity (KD 1.04 +/- 0.23 nM) to a single class of sites, of which there were approximately 200,000/cell. In competition experiments antagonist binding was generally homogeneous, whereas that of agonists was heterogeneous and subpopulations of binding sites with different affinities for agonists were identified. The Ki value of 8.1 +/- 1.1 nM for inhibition of QNB binding by atropine was greater than the pA2 of 10.2 +/- 1.1 derived from contraction studies. Furthermore, whereas picomolar concentrations of agonists induced cell contraction, substantially higher concentrations (10 nM to 10 mM) were required to inhibit [3H]QNB binding to the isolated cells.

Transmembrane 45Ca fluxes in isolated smooth muscle cells: basal Ca2+ fluxes

1984 ◽  
Vol 246 (5) ◽  
pp. C422-C430 ◽  
Author(s):  
C. R. Scheid ◽  
F. S. Fay
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Slow Component ◽  
Compartmental Analysis ◽  
Muscle Cells ◽  
Stomach Muscle ◽  
Cellular Processes ◽  
Transmembrane Flux ◽  
45Ca Efflux ◽  
Isolated Smooth Muscle Cells

Methods are described for monitoring unidirectional fluxes of 45Ca in smooth muscle cells in suspension. Compartmental analysis of 45Ca influx data indicate that 45Ca exchange consists of two kinetically distinguishable components in the toad stomach muscle cells: a relatively small (approximately 100-pmol X cm-2) component with a rapid rate of exchange (t1/2 approximately 1.4 min) and a larger (approximately 930-pmol X cm-2) component, which exchanges more slowly (t1/2 congruent to 87 min). The rate of exchange of the latter but not the former Ca2+ pool is increased on exposure of the cells to elevated K+ levels; thus the “slow” component of uptake appears to reflect transmembrane Ca2+ flux, whereas the “rapid” component may reflect exchange of surface-bound label. Consistent with this interpretation is the finding that under conditions in which surface 45Ca is rapidly and completely displaced, 45Ca efflux occurs as a simple monoexponential process with a slow rate of exchange (k = 7.80 X 10(-5) s-1). The apparent rate of transmembrane Ca2+ flux in smooth muscle cells at rest is approximately 0.1 pmol X cm-2 X s-1. Cellular processes that could give rise to an apparent transmembrane flux rate of this magnitude are discussed, and a model is presented which appears to describe cellular 45Ca exchange in the isolated smooth muscle cells.

In vivo membrane potentials of smooth muscle cells in the caudal artery of the rat

1985 ◽  
Vol 249 (1) ◽  
pp. C78-C83 ◽  
Author(s):  
H. J. Bryant ◽  
D. R. Harder ◽  
M. B. Pamnani ◽  
F. J. Haddy
Keyword(s):  
Smooth Muscle ◽  
Membrane Potential ◽  
Smooth Muscle Cells ◽  
Muscle Cells ◽  
Membrane Potentials ◽  
Resting Membrane Potential ◽  
Humoral Factors ◽  
Caudal Artery ◽  
Blood Pressures

Membrane potentials measured in vivo may differ significantly from those measured in vitro in part due to humoral factors, innervation, and wall tension. These studies were initiated to determine whether it is feasible to record membrane potentials from vascular smooth muscle cells in vivo in the caudal artery of the pentobarbital-anesthetized male Wistar rat. Membrane potentials were measured using glass microelectrodes and correlated with systolic, diastolic, and mean blood pressures. For systolic blood pressures between 100 and 140 mmHg the average resting membrane potential was -38.4 +/- 0.48 mV. There was good correlation of systolic, diastolic, and mean blood pressures with membrane potential between 100 and 140 mmHg (r = 0.89, 0.75, and 0.89, respectively). Below 80 mmHg the arterial muscle cells became more depolarized than would be expected if the membrane potential were determined solely by transmural pressure. The depolarized membrane potential at low arterial pressures may be due to enhanced neural input. Spontaneous electrical activity was observed in some of the in vivo cells. When action potentials were present, they were generated at rates between 1-2/s and 6-7/min. These studies indicate that it is feasible to measure membrane potentials from arterial smooth muscle cells in vivo in the caudal artery of the rat.

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

2015 ◽  
Vol 308 (7) ◽  
pp. G625-G633 ◽  
Author(s):  
Ji Yeon Lee ◽  
Eun-ju Ko ◽  
Ki Duck Ahn ◽  
Sung Kim ◽  
Poong-Lyul Rhee
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Smooth Muscles ◽  
Muscle Cells ◽  
Bk Channels ◽  
Resting Membrane Potential ◽  
Membrane Excitability ◽  
Gastric Corpus ◽  
Isolated Smooth Muscle Cells

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.

Rectifying properties of the membrane of single freshly isolated smooth muscle cells

1980 ◽  
Vol 239 (5) ◽  
pp. C175-C181 ◽  
Author(s):  
J. J. Singer ◽  
J. V. Walsh
Keyword(s):  
Smooth Muscle ◽  
Membrane Potential ◽  
Smooth Muscle Cells ◽  
Fold Increase ◽  
Muscle Cells ◽  
Tetraethylammonium Chloride ◽  
Intracellular Microelectrodes ◽  
Toad Bufo ◽  
Isolated Smooth Muscle Cells ◽  
Direct Microscopic Observation

Single smooth muscle cells freshly isolated from the stomach muscularis of the toad Bufo marinus were studied under direct microscopic observation using intracellular microelectrodes. The deviation of the membrane potential from rest was recorded when steps of current were injected into the cell. Outward-going rectification was consistently observed both in the presence of 1.8 mM and higher external concentrations of Ca2+. There was no indication of inward-going rectification even under conditions favoring its demonstration, i.e., when the external concentration of K+ was high (108 mM) and Cl-, low (39.6 mM). In the presence of tetraethylammonium chloride (TEA), there was a marked decrease in the rectification normally observed with depolarizing currents, suggesting that a K+ conductance contributes to the outward-going rectification. This K+ conductance increased by almost two orders of magnitude over the range from -20 to 0 mV, and displayed an e-fold increase with a depolarization as small as 4-7 mV. In response to hyperpolarizing currents, the membrane potential did not always reach a plateau but at times continued to become more negative. The feasibility of the depletion of ions from the caveolae as an explanation for this observation is discussed.

Chronic carbon monoxide enhanced IbTx-sensitive currents in rat resistance pulmonary artery smooth muscle cells

2002 ◽  
Vol 283 (1) ◽  
pp. L120-L129 ◽  
Author(s):  
Eric Dubuis ◽  
Mathieu Gautier ◽  
Alexandre Melin ◽  
Manuel Rebocho ◽  
Catherine Girardin ◽  
...  
Keyword(s):  
Carbon Monoxide ◽  
Smooth Muscle ◽  
Membrane Potential ◽  
Pulmonary Artery ◽  
Smooth Muscle Cells ◽  
Outward Current ◽  
Muscle Cells ◽  
Membrane Resistance ◽  
Resting Membrane Potential ◽  
Whole Cell Patch Clamp

Exogenous carbon monoxide (CO) can induce pulmonary vasodilation by acting directly on pulmonary artery (PA) smooth muscle cells. We investigated the contribution of K+ channels to the regulation of resistance PA resting membrane potential on control (PAC) rats and rats exposed to CO for 3 wk at 530 parts/million, labeled as PACO rats. Whole cell patch-clamp experiments revealed that the resting membrane potential of PACO cells was more negative than that of PAC cells. This was associated with a decrease of membrane resistance in PACO cells. Additional analysis showed that outward current density in PACO cells was higher (50% at +60 mV) than in PAC cells. This was linked to an increase of iberiotoxin (IbTx)-sensitive current. Chronic CO hyperpolarized membrane of pressurized PA from −46.9 ± 1.2 to −56.4 ± 2.6 mV. Additionally, IbTx significantly depolarized membrane of smooth muscle cells from PACO arteries but not from PAC arteries. The present study provides initial evidence of an increase of Ca2+-activated K+ current in smooth muscle cells from PA of rats exposed to chronic CO.

Membrane properties of smooth muscle cells in pulmonary hypertensive rats

1982 ◽  
Vol 242 (5) ◽  
pp. H907-H915 ◽  
Author(s):  
H. Suzuki ◽  
B. M. Twarog
Keyword(s):  
Smooth Muscle ◽  
Membrane Potential ◽  
Pulmonary Artery ◽  
Smooth Muscle Cells ◽  
Main Pulmonary Artery ◽  
Fold Increase ◽  
Muscle Cells ◽  
Membrane Properties ◽  
Resting Membrane Potential ◽  
Chronic Pulmonary Hypertension

The membrane properties of smooth muscle cells in rat main pulmonary artery (MPA) and small pulmonary artery (SPA) were investigated during chronic normobaric hypoxia and after monocrotaline injection. As chronic pulmonary hypertension developed, pronounced differences between MPA and SPA were observed. These findings may shed light on mechanisms of smooth muscle hypertrophy. 1) The resting membrane potential of smooth muscle in MPA became less negative than the normal (depolarized), whereas the resting membrane potential of smooth muscle in SPA became more negative (hyperpolarized). 2) In MPA, both the length and time constants diminished. 3) In MPA, the maximum membrane depolarization produced by a 10-fold increase in extracellular [K+] decreased. 4) In SPA, the depolarization observed in K+-free solution was more rapid and greater in amplitude, and the transient hyperpolarization following restoration of K+-containing solution increased. 5) In SPA, initial and sustained depolarization evoked by Na+-deficient solutions were increased. 6) Depolarization in MPA was due to increased membrane permeability, perhaps to Cl-, whereas hyperpolarization in SPA could be attributed to increased activity of an electrogenic Na+-K+ pump.

Pharmacological and biophysical isolation of K+ currents encoded by ether-à-go-go-related genes in murine hepatic portal vein smooth muscle cells

2007 ◽  
Vol 292 (1) ◽  
pp. C468-C476 ◽  
Author(s):  
Shuk Yin M. Yeung ◽  
Iain A. Greenwood
Keyword(s):  
Smooth Muscle ◽  
Membrane Potential ◽  
Portal Vein ◽  
Smooth Muscle Cells ◽  
Significant Proportion ◽  
Muscle Cells ◽  
Resting Membrane Potential ◽  
Perforated Patch ◽  
Inward Currents ◽  
Channel Currents

Previous studies have shown that murine portal vein myocytes express ether-à-go-go related genes (ERGs) and exhibit distinctive currents when recorded under symmetrical K+ conditions. The aim of the present study was to characterize ERG channel currents evoked from a negative holding potential under conditions more pertinent to a physiological scenario to assess the possible functional impact of this conductance. Currents were recorded with ruptured or perforated patch variants of the whole cell technique from a holding potential of −60 mV. Application of three structurally distinct and selective ERG channel blockers, E-4031, dofetilide, and the peptide toxin BeKM-1, all inhibited a significant proportion of the outward current and abolished inward currents with distinctive “hooked” kinetics recorded on repolarization. Dofetilide-sensitive currents at negative potentials evoked by depolarization to +40 mV had a voltage-dependent time to peak and rate of decay characteristic of ERG channels. Application of the novel ERG channel activator PD-118057 (1–10 μM) markedly enhanced the hooked inward currents evoked by membrane depolarization and hyperpolarized the resting membrane potential recorded by current clamp and the perforated patch configuration by ∼20 mV. In contrast, ERG channel blockade by dofetilide (1 μM) depolarized the resting membrane potential by ∼8 mV. These data are the first record of ERG channel currents in smooth muscle cells under quasi-physiological conditions that suggest that ERG channels contribute to the resting membrane potential in these cells.

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