Differential effects of soluble and particulate guanylyl cyclase on Ca2+ sensitivity in airway smooth muscle

2002 ◽  
Vol 92 (1) ◽  
pp. 257-263 ◽  
Author(s):  
Edwin H. Rho ◽  
William J. Perkins ◽  
Robert R. Lorenz ◽  
David O. Warner ◽  
Keith A. Jones
Keyword(s):  
Smooth Muscle ◽  
Airway Smooth Muscle ◽  
Guanylyl Cyclase ◽  
Soluble Guanylyl Cyclase ◽  
Rightward Shift ◽  
Muscarinic Stimulation ◽  
Intracellular Ca ◽  
A Cell ◽  
The Relationship ◽  
Stimulation Of

Maximal relaxation of airway smooth muscle (ASM) in response to atrial natriuretic peptide (ANP), which stimulates particulate guanylyl cyclase (pGC), is less than that produced by nitric oxide (NO) and other compounds that stimulate soluble guanylyl cyclase (sGC). We hypothesized that stimulation of pGC relaxes ASM only by decreasing intracellular Ca2+ concentration ([Ca2+]i), whereas stimulation of sGC decreases both [Ca2+]i and the force developed for a given [Ca2+]i (i.e., the Ca2+ sensitivity) during muscarinic stimulation. We measured the relationship between force and [Ca2+]i (using fura 2) under control conditions (using diltiazem to change [Ca2+]i) and during exposure to ANP, diethylamine-NO (DEA-NO), sodium nitroprusside (SNP), and the Sp diastereoisomer of β-phenyl-1, N 2-etheno-8-bromoguanosine-3′,5′-cyclic monophosphorothionate ( Sp-8-Br-PET-cGMPS), a cell-permeant analog of cGMP. Addition of DEA-NO, SNP, or Sp-8-Br-PET-cGMPS decreased both [Ca2+]i and force, causing a significant rightward shift of the force-[Ca2+]irelationship. In contrast, with ANP exposure, the force-[Ca2+]i relationship was identical to control, such that ANP produced relaxation solely by decreasing [Ca2+]i. Thus, during muscarinic stimulation, stimulation of pGC relaxes ASM exclusively by decreasing [Ca2+]i, whereas stimulation of sGC decreases both [Ca2+]i and Ca2+sensitivity.

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

Relationship between force and regulatory myosin light chain phosphorylation in airway smooth muscle

2000 ◽  
Vol 279 (1) ◽  
pp. L52-L58 ◽  
Author(s):  
Tetsuya Kai ◽  
Hayashi Yoshimura ◽  
Keith A. Jones ◽  
David O. Warner
Keyword(s):  
Smooth Muscle ◽  
Light Chain ◽  
Myosin Light Chain ◽  
Myosin Light Chain Phosphorylation ◽  
Mean Values ◽  
Regulatory Myosin Light Chain ◽  
Muscarinic Stimulation ◽  
The Relationship ◽  
Force Relationship ◽  
Stimulation Of

We tested the hypothesis that increases in force at a given cytosolic Ca2+ concentration (i.e., Ca2+ sensitization) produced by muscarinic stimulation of canine tracheal smooth muscle (CTSM) are produced in part by mechanisms independent of changes in regulatory myosin light chain (rMLC) phosphorylation. This was accomplished by comparing the relationship between rMLC phosphorylation and force in α-toxin-permeabilized CTSM in the absence and presence of acetylcholine (ACh). Forces were normalized to the contraction induced by 10 μM Ca2+ in each strip, and rMLC phosphorylation is expressed as a percentage of total rMLC. ACh (100 μM) plus GTP (1 μM) significantly shifted the Ca2+-force relationship curve to the left (EC50: 0.39 ± 0.06 to 0.078 ± 0.006 μM Ca2+) and significantly increased the maximum force (104.4 ± 4.8 to 120.2 ± 2.8%; n = 6 observations). The Ca2+-rMLC phosphorylation relationship curve was also shifted to the left (EC50: 1.26 ± 0.57 to 0.13 ± 0.04 μM Ca2+) and upward (maximum rMLC phosphorylation: 70.9 ± 7.9 to 88.5 ± 5.1%; n = 6 observations). The relationships between rMLC phosphorylation and force constructed from mean values at corresponding Ca2+concentrations were not different in the presence and absence of ACh. We find no evidence that muscarinic stimulation increases Ca2+ sensitivity in CTSM by mechanisms other than increases in rMLC phosphorylation.

Halothane Increases Smooth Muscle Protein Phosphatase in Airway Smooth Muscle

2001 ◽  
Vol 94 (1) ◽  
pp. 129-136 ◽  
Author(s):  
Motohiko Hanazaki ◽  
Keith A. Jones ◽  
William J. Perkins ◽  
David O. Warner
Keyword(s):  
Smooth Muscle ◽  
Airway Smooth Muscle ◽  
Light Chain ◽  
Protein Phosphatase ◽  
Myosin Light Chain ◽  
Muscle Protein ◽  
Isometric Force ◽  
Activity Assay ◽  
Muscarinic Stimulation ◽  
Intracellular Ca

Background Halothane relaxes airway smooth muscle, in part, by decreasing the force produced for a given intracellular [Ca(2+)] (i.e., Ca(2+) sensitivity) during muscarinic stimulation, an effect produced by a decrease in regulatory myosin light-chain (rMLC) phosphorylation. The authors tested the hypothesis that halothane reduces rMLC phosphorylation during muscarinic stimulation at constant intracellular [Ca(2+)] by increasing smooth muscle protein phosphatase (SMPP) activity, without changing myosin light-chain kinase (MLCK) activity. Methods Enzyme activities were assayed in beta-escin permeabilized strips of canine tracheal smooth muscle. Under conditions of constant intracellular [Ca(2+)], the rate of rMLC phosphorylation was measured by Western blotting during inhibition of SMPP with microcystin-LR (to assay MLCK activity) or during inhibition of MLCK by wortmannin and adenosine triphosphate depletion (to assay SMPP activity). The effect of halothane (0.8 mm) on enzyme activities and isometric force during stimulation with 0.6 microm Ca(2+) and 10 microm acetylcholine was determined. Results Halothane produced a 14 +/- 8% (mean +/- SD) decrease in isometric force by significantly reducing rMLC phosphorylation (from 32 +/- 9% to 28 +/- 9%). Halothane had no significant effect on any parameter of a monoexponential relation fit to the data for the MLCK activity assay. In contrast, halothane significantly decreased the half-time for rMLC dephosphorylation in the SMPP activity assay (from 0.74 +/- 0.28 min to 0.44 +/- 0.10 min), indicating that it increased SMPP activity. Conclusions Halothane decreases Ca(2+) sensitivity and rMLC phosphorylation in airway smooth muscle during muscarinic receptor stimulation by increasing SMPP activity, without affecting MLCK, probably by disrupting receptor G-protein signaling pathways that inhibit SMPP.

scholarly journals Soluble Guanylyl Cyclase is Reduced in Airway Smooth Muscle Cells From a Murine Model of Allergic Asthma

2010 ◽  
Vol 3 (12) ◽  
pp. 271-276
Author(s):  
Fabiola Placeres-Uray ◽  
Ramona González de Alfonzo ◽  
Itala Lippo de Becemberg ◽  
Marcelo J. Alfonzo
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Allergic Asthma ◽  
Airway Smooth Muscle ◽  
Guanylyl Cyclase ◽  
Murine Model ◽  
Soluble Guanylyl Cyclase ◽  
Muscle Cells ◽  
Airway Smooth Muscle Cells

Effects of salbutamol on intracellular calcium oscillations in porcine airway smooth muscle

1997 ◽  
Vol 82 (6) ◽  
pp. 1836-1843 ◽  
Author(s):  
Y. S. Prakash ◽  
H. F. M. Van Der Heijden ◽  
M. S. Kannan ◽  
G. C. Sieck
Keyword(s):  
Smooth Muscle ◽  
Intracellular Calcium ◽  
Airway Smooth Muscle ◽  
Inhibitory Effect ◽  
Calcium Oscillations ◽  
Adrenoceptor Agonist ◽  
Intracellular Ca ◽  
Adrenoceptor Agonists ◽  
A Cell ◽  
Agonist Concentration

Prakash, Y. S., H. F. M. van der Heijden, M. S. Kannan, and G. C. Sieck. Effects of salbutamol on intracellular calcium oscillations in porcine airway smooth muscle. J. Appl. Physiol. 82(6): 1836–1843, 1997.—Relaxation of airway smooth muscle (ASM) by β-adrenoceptor agonists involves reduction of intracellular Ca2+concentration ([Ca2+]i). In porcine ASM cells, acetylcholine induces [Ca2+]ioscillations that display frequency modulation by agonist concentration and basal [Ca2+]i. We used real-time confocal microscopy to examine the effect of salbutamol (1 nM to 1 μM), a β2-adrenoceptor agonist, on [Ca2+]ioscillations in freshly dissociated porcine ASM cells. Salbutamol decreased the frequency of [Ca2+]ioscillations in a concentration-dependent fashion, completely inhibiting the oscillations at 1 μM. These effects were mimicked by a cell-permeant analog of adenosine 3′,5′-cyclic monophosphate. The inhibitory effect of salbutamol was partially reversed by BAY K 8644. Salbutamol reduced [Ca2+]ieven when sarcoplasmic reticulum (SR) Ca2+ reuptake and Ca2+ influx were blocked. Lanthanum blockade of Ca2+ efflux attenuated the inhibitory effect of salbutamol on [Ca2+]i. The [Ca2+]iresponse to caffeine was unaffected by salbutamol. On the basis of these results, we conclude that β2-adrenoceptor agonists have little effect on SR Ca2+ release in ASM cells but reduce [Ca2+]iby inhibiting Ca2+ influx through voltage-gated channels and by enhancing Ca2+ efflux.

scholarly journals Glutamate regulates Ca2+ signals in smooth muscle cells of newborn piglet brain slice arterioles through astrocyte- and heme oxygenase-dependent mechanisms

2010 ◽  
Vol 298 (2) ◽  
pp. H562-H569 ◽  
Author(s):  
Qi Xi ◽  
Edward Umstot ◽  
Guiling Zhao ◽  
Damodaran Narayanan ◽  
Charles W. Leffler ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Heme Oxygenase ◽  
Guanylyl Cyclase ◽  
Brain Slices ◽  
Soluble Guanylyl Cyclase ◽  
Muscle Cells ◽  
Newborn Piglet ◽  
Signal Modulation ◽  
Cerebral Arterioles

Glutamate is the principal cerebral excitatory neurotransmitter and dilates cerebral arterioles to match blood flow to neural activity. Arterial contractility is regulated by local and global Ca2+ signals that occur in smooth muscle cells, but modulation of these signals by glutamate is poorly understood. Here, using high-speed confocal imaging, we measured the Ca2+ signals that occur in arteriole smooth muscle cells of newborn piglet tangential brain slices, studied signal regulation by glutamate, and investigated the physiological function of heme oxygenase (HO) and carbon monoxide (CO) in these responses. Glutamate elevated Ca2+ spark frequency by ∼188% and reduced global intracellular Ca2+ concentration ([Ca2+]i) to ∼76% of control but did not alter Ca2+ wave frequency in brain arteriole smooth muscle cells. Isolation of cerebral arterioles from brain slices abolished glutamate-induced Ca2+ signal modulation. In slices treated with l-2-α-aminoadipic acid, a glial toxin, glutamate did not alter Ca2+ sparks or global [Ca2+]i but did activate Ca2+ waves. This shift in Ca2+ signal modulation by glutamate did not occur in slices treated with d-2-α-aminoadipic acid, an inactive isomer of l-2-α-aminoadipic acid. In the presence of chromium mesoporphyrin, a HO blocker, glutamate inhibited Ca2+ sparks and Ca2+ waves and did not alter global [Ca2+]i. In isolated arterioles, CORM-3 [tricarbonylchloro(glycinato)ruthenium(II)], a CO donor, activated Ca2+ sparks and reduced global [Ca2+]i. These effects were blocked by 1 H-(1,2,4)-oxadiazolo-(4,3-a)-quinoxalin-1-one, a soluble guanylyl cyclase inhibitor. Collectively, these data indicate that glutamate can modulate Ca2+ sparks, Ca2+ waves, and global [Ca2+]i in arteriole smooth muscle cells via mechanisms that require astrocytes and HO. These data also indicate that soluble guanylyl cyclase is involved in CO activation of Ca2+ sparks in arteriole smooth muscle cells.

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)

Sign in / Sign up

Export Citation Format

Share Document