Src modulates serotonin-induced calcium signaling by regulating phosphatidylinositol 4,5-bisphosphate

2002 ◽  
Vol 282 (6) ◽  
pp. L1305-L1313 ◽  
Author(s):  
Barbara Tolloczko ◽  
Petra Turkewitsch ◽  
Sofia Choudry ◽  
Sandra Bisotto ◽  
Elizabeth D. Fixman ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Inositol Phosphates ◽  
Src Kinase ◽  
Muscle Cells ◽  
Tracheal Smooth Muscle ◽  
Airway Smooth Muscle Cells ◽  
Cell Contractility ◽  
Rat Tracheal Smooth Muscle ◽  
Stimulation Of

We tested the hypothesis that, in airway smooth muscle cells, stimulation of G-protein-coupled receptors by contractile agonists activates Src kinase and that this kinase modulates cell contractility and Ca2+ signaling by affecting the levels of the phospholipase C substrate phosphatidylinositol 4,5-bisphosphate (PIP2). Stimulation of cultured rat tracheal smooth muscle cells with serotonin (5-HT) induced an increase in Src activity, Ca2+ mobilization, and contraction (decrease in cell area). 5-HT-evoked cell contraction was reduced by a specific inhibitor of Src family kinases, 4-amino-5(4-methylphenyl)-7-( t-butyl)pyrazolo[3,4 -d]pyrimidine (PP1). Peak Ca2+ responses to 5-HT were attenuated by PP1 and an anti-Src-blocking antibody and augmented by expression of constitutively activated Y529F Src. Sustained phases of Ca2+ responses to 5-HT and Ca2+ influx resulting from emptying of Ca2+ stores in the endoplasmic reticulum by thapsigargin were also decreased after PP1 treatment. PP1 significantly reduced the turnover of inositol phosphates produced on 5-HT stimulation and the amount of PIP2 in the Triton X-100-insoluble lipid fraction. Overall, these data demonstrate that, in rat tracheal smooth muscle cells, Src kinase modulates 5-HT-evoked cell contractility and Ca2+ signaling by regulating PIP2 levels and Ca2+ influx.

Hyperoxia inhibits proliferation of cultured rat tracheal smooth muscle cells

1994 ◽  
Vol 267 (2) ◽  
pp. L101-L105 ◽  
Author(s):  
M. Absher ◽  
W. Makrides ◽  
P. Shapiro ◽  
J. N. Evans
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Protein Content ◽  
Muscle Cells ◽  
Tracheal Smooth Muscle ◽  
Lung Fibroblasts ◽  
Airway Smooth Muscle Cells ◽  
Specific Cell ◽  
Isolated Cells ◽  
Rat Tracheal Smooth Muscle

Exposure of the lung to elevated oxygen leads to structural and cellular injury followed by extensive tissue remodeling. In vitro models utilizing isolated cells exposed to hyperoxic conditions or exogenously added oxidants may be injurious or stimulatory depending on the specific cell type and level and duration of exposure. In the present study, proliferation of cultured rat tracheal smooth muscle cells was inhibited by oxygen concentrations of 40 and 70% compared with a "normoxic" concentration of 21%. Exposure to 70% oxygen had a hypertrophic effect on the cells, as indicated by increased cellular protein content, whereas cells exposed to 21% oxygen did not show increased protein content. Exogenously added oxidant, H2O2, resulted in complete inhibition of growth of tracheal smooth muscle cells at concentrations > 3 microM. Much higher concentrations of H2O2 were required to inhibit proliferation of vascular smooth muscle cells and rat lung fibroblasts. The heightened sensitivity of airway smooth muscle cells to oxygen and oxidants may be an important factor in the early events following hyperoxia-induced lung injury.

Effect of the cytoskeletal prestress on the mechanical impedance of cultured airway smooth muscle cells

2002 ◽  
Vol 92 (4) ◽  
pp. 1443-1450 ◽  
Author(s):  
Dimitrije Stamenović ◽  
Zhuangli Liang ◽  
Jianxin Chen ◽  
Ning Wang
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Airway Smooth Muscle ◽  
Mechanical Impedance ◽  
Muscle Cells ◽  
Airway Smooth Muscle Cells ◽  
Integrin Receptors ◽  
Cell Contractility ◽  
Frictional Properties ◽  
Applied Torque

We investigated the effect of the cytoskeletal prestress ( P) on the elastic and frictional properties of cultured human airway smooth muscle cells during oscillatory loading; P is preexisting tensile stress in the actin cytoskeleton generated by the cell contractile apparatus. We oscillated (0.1 Hz, 6 Pa peak to peak) small ferromagnetic beads bound to integrin receptors and computed the storage (elastic) modulus ( G′) and the loss (frictional) modulus ( G") from the applied torque and the corresponding bead rotation. All measurements were done at baseline and after cells were treated with graded doses of either histamine (0.1, 1, 10 μM) or isoproterenol (0.01, 0.1, 1, 10 μM). Values for P for these concentrations were taken from a previous study (Wang et al., Am J Physiol Cell Physiol, in press). It was found that G′ and G", as well as P, increased/decreased with increasing doses of histamine/isoproterenol. Both G′ and G" exhibited linear dependences on P: G′(Pa) = 0.20 P + 82 and G"(Pa) = 0.05 P + 32. The dependence of G′ on P is consistent with our previous findings and with the behavior of stress-supported structures. The dependence of G" on P is a novel finding. It could be attributed to a variety of mechanisms. Some of those mechanisms are discussed in detail. We concluded that, in addition to the central mechanisms by which stress-supported structures develop mechanical stresses, other mechanisms might need to be invoked to fully explain the observed dependence of the cell mechanical properties on the state of cell contractility.

Muscarinic stimulation of tracheal smooth muscle cells activates large-conductance Ca2+-dependent K+ channel

1997 ◽  
Vol 272 (3) ◽  
pp. 1-1 ◽  
Author(s):  
G. R. Wade ◽  
S. M. Sims
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Muscle Cells ◽  
Tracheal Smooth Muscle ◽  
K Channel ◽  
Muscarinic Stimulation ◽  
Cholinergic Response ◽  
Stimulation Of

Pages C658-C665: G. R. Wade and S. M. Sims. “Muscarinic stimulation of tracheal smooth muscle cells activates large-conductance Ca2+-dependent K+ channel.” Page C662, Fig. 6: the second half of the trace in A was inadvertently duplicated from B. The revised Fig. 6 below shows the correct channel traces. We wish to emphasize that the scientific point of the figure, the reversible antagonism of the cholinergic response by atropine, as well as the quantification in D, remains sound. The data were filtered at 400 Hz and sampled off-line from digital videotape at 2 kHz. (See PDF)

Muscarinic stimulation of tracheal smooth muscle cells activates large-conductance Ca(2+)-dependent K+ channel

1993 ◽  
Vol 265 (3) ◽  
pp. C658-C665 ◽  
Author(s):  
G. R. Wade ◽  
S. M. Sims
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Muscle Cells ◽  
Tracheal Smooth Muscle ◽  
Equilibrium Potential ◽  
Channel Activity ◽  
Unitary Conductance ◽  
Channel Opening ◽  
Muscarinic Stimulation ◽  
Stimulation Of

We investigated the regulation of the large-conductance Ca(2+)-dependent K+ (KCa) channel by acetylcholine (ACh) in freshly dissociated tracheal smooth muscle cells. Channels were recorded in the cell-attached patch configuration, and cells were stimulated with ACh, muscarine, or caffeine. We identified KCa channel activity based on 1) the voltage dependence of channel opening; 2) the large unitary conductance (242 +/- 5 pS with symmetrical 135 mM K+); 3) dependence of the reversal potential on the [K+] gradient, shifting 56 +/- 3 mV/10-fold change in extracellular [K+]; and 4) opening of channels after elevation of cytosolic free Ca2+ concentration ([Ca2+]i) using the Ca2+ ionophore A23187. When cells were bathed either in a physiological saline solution or a solution containing 135 mM K+ (to clamp cell membrane potential near 0 mV), ACh caused contraction of cells and activation of voltage-dependent channels. With 135 mM extracellular K+, the channels activated by ACh had a unitary conductance of 247 +/- 10 pS, and currents reversed near the K+ equilibrium potential (-1 +/- 1 mV). The effects of ACh were reversible, blocked by atropine, and mimicked by muscarine. From these characteristics we conclude that muscarinic stimulation of canine tracheal smooth muscle cells leads to activation of the large-conductance KCa channel. Because the KCa channels were isolated from ACh by the patch pipette, the increased channel activity was probably mediated by a cytosolic second messenger. ACh shifted the threshold for KCa channel opening to less positive membrane potentials, similar to that seen with elevation of [Ca2+]i.(ABSTRACT TRUNCATED AT 250 WORDS)

Muscarinic Stimulation of Airway Smooth Muscle Cells

1998 ◽  
Vol 31 (3) ◽  
pp. 349-356 ◽  
Author(s):  
Etienne Roux ◽  
Mathieu Molimard ◽  
Savineau ◽  
Roger Marthan
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Airway Smooth Muscle ◽  
Muscle Cells ◽  
Airway Smooth Muscle Cells ◽  
Muscarinic Stimulation ◽  
Stimulation Of

Serotonin-evoked calcium transients in airway smooth muscle cells

1995 ◽  
Vol 269 (2) ◽  
pp. L234-L240 ◽  
Author(s):  
B. Tolloczko ◽  
Y. L. Jia ◽  
J. G. Martin
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Muscle Cells ◽  
Airway Smooth Muscle Cells ◽  
Second Phase ◽  
Receptor Subtypes ◽  
Airway Narrowing ◽  
Airway Responses ◽  
Rat Tracheal Smooth Muscle ◽  
In Cells

Serotonin (5-HT) is an important mediator of allergic airway narrowing in several animal species. The present study was designed to characterize the receptor subtypes that mediate 5-HT-induced airway responses in the rat. To do this, we measured Ca2+ transients and adenosine 3',5'-cyclic monophosphate (cAMP) production evoked by 5-HT in cultured rat tracheal smooth muscle cells as well as 5-HT-induced contractions of isolated tracheal rings. 5-HT (10(-6) to 10(-4) M) triggered a rapid increase in intracellular Ca2+ concentration ([Ca2+]i) followed by a second phase of sustained, elevated, and sometimes oscillating levels of [Ca2+]i. Sustained but not peak [Ca2+]i levels were dependent on Ca2+ influx but were not attenuated by nifedipine (10(-5) M). Oscillations were observed in cells in Ca2+ -free medium, suggesting Ca2+ -induced Ca2+ release independent of Ca2+ influx. The effects of 5-HT were inhibited by thapsigargin (10(-6) M) and ketanserin (10(-7) M). In cells incubated with LY-278,584 (5-HT3 antagonist) and (-)pindolol (5-HT1 antagonist), 5-HT-evoked responses were not significantly different from the control values. 5-methyltryptamine (5-MT), a ligand with higher affinity for 5-HT2C receptors than "classical" 5-HT2 receptors, elicited higher responses than dipropyl-5-carboxamidotryptamine (DP-5-CT), which possesses lower affinity for 5-HT2C receptors than 5-MT, but an affinity for the classical 5-HT2 receptor similar to 5-MT, but an affinity for the classical 5-HT2 receptor similar to 5-MT.(ABSTRACT TRUNCATED AT 250 WORDS)

Muscarinic receptor stimulation modulates the effect of halothane on Mn2+ influx in airway smooth muscle

1997 ◽  
Vol 273 (3) ◽  
pp. C868-C873 ◽  
Author(s):  
D. O. Warner ◽  
K. A. Jones ◽  
R. R. Lorenz ◽  
C. M. Pabelick
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Airway Smooth Muscle ◽  
Muscle Cells ◽  
Tracheal Smooth Muscle ◽  
Airway Smooth Muscle Cells ◽  
Maximal Stimulation ◽  
Muscarinic Stimulation ◽  
Dose Dependent Increase ◽  
Dose Dependent

Prior studies suggest that the mechanism of action by which halothane relaxes airway smooth muscle depends on the contractile state of the cell. We hypothesized that halothane would inhibit the influx of Ca2+ into canine airway smooth muscle cells during submaximal, but not maximal, muscarinic stimulation. This hypothesis was tested by using the rate of quenching of fura 2 fluorescence by Mn2+ in strips of canine tracheal smooth muscle as an index of Ca2+ influx. Acetylcholine (ACh) produced a dose-dependent increase in Mn2+ influx. Halothane (0.64 +/- 0.05 microM) significantly decreased Mn2+ influx and intracellular Ca2+ concentration when added to strips stimulated with a submaximal concentration of ACh (0.3 microM) but had no effect on Mn2+ influx or intracellular Ca2+ concentration during maximal stimulation with ACh (100 microM). Similar results were observed when the strips were treated with verapamil. These results demonstrate that anesthetic effects on Ca2+ homeostasis in intact canine tracheal smooth muscle cells may be critically modulated by receptor-linked mechanisms.

Endothelin-1 Regulates Proliferative Responses, both alone and Synergistically with PDGF, in Rat Tracheal Smooth Muscle Cells

2006 ◽  
Vol 17 (1-2) ◽  
pp. 37-46 ◽  
Author(s):  
Linda Yahiaoui ◽  
Annie Villeneuve ◽  
Héctor Valderrama-Carvajal ◽  
Fiona Burke ◽  
Elizabeth D. Fixman
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Muscle Cells ◽  
Tracheal Smooth Muscle ◽  
Endothelin 1 ◽  
Rat Tracheal Smooth Muscle
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