Relationship between in vivo Airway Reactivity and in vitro Responsiveness of Tracheal Smooth Muscle in Inbred Rats

Respiration ◽  
1988 ◽  
Vol 54 (1) ◽  
pp. 108-113 ◽  
Author(s):  
G.U. Di Maria ◽  
J.G. Martin ◽  
S. Bellofiore ◽  
A. Mistretta
Keyword(s):  
Smooth Muscle ◽  
Tracheal Smooth Muscle ◽  
Airway Reactivity ◽  
Inbred Rats

Phosphorylation of caldesmon by ERK MAP kinases in smooth muscle

2000 ◽  
Vol 278 (4) ◽  
pp. C718-C726 ◽  
Author(s):  
Jason C. Hedges ◽  
Brian C. Oxhorn ◽  
Michael Carty ◽  
Leonard P. Adam ◽  
Ilia A. Yamboliev ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Map Kinase ◽  
Map Kinases ◽  
Isometric Force ◽  
Phosphorylation Site ◽  
Smooth Muscle Contraction ◽  
Tracheal Smooth Muscle ◽  
Muscarinic Agonists

Phosphorylation of h-caldesmon has been proposed to regulate airway smooth muscle contraction. Both extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein (MAP) kinases phosphorylate h-caldesmon in vitro. To determine whether both enzymes phosphorylate caldesmon in vivo, phosphorylation-site-selective antibodies were used to assay phosphorylation of MAP kinase consensus sites. Stimulation of cultured tracheal smooth muscle cells with ACh or platelet-derived growth factor increased caldesmon phosphorylation at Ser789 by about twofold. Inhibiting ERK MAP kinase activation with 50 μM PD-98059 blocked agonist-induced caldesmon phosphorylation completely. Inhibiting p38 MAP kinases with 25 μM SB-203580 had no effect on ACh-induced caldesmon phosphorylation. Carbachol stimulation increased caldesmon phosphorylation at Ser789 in intact tracheal smooth muscle, which was blocked by the M2 antagonist AF-DX 116 (1 μM). AF-DX 116 inhibited carbachol-induced isometric contraction by 15 ± 1.4%, thus dissociating caldesmon phosphorylation from contraction. Activation of M2 receptors leads to activation of ERK MAP kinases and phosphorylation of caldesmon with little or no functional effect on isometric force. P38 MAP kinases are also activated by muscarinic agonists, but they do not phosphorylate caldesmon in vivo.

Release of epithelium-derived PGE2 from canine trachea after antigen inhalation

1998 ◽  
Vol 274 (2) ◽  
pp. L220-L225 ◽  
Author(s):  
I. McGrogan ◽  
L. J. Janssen ◽  
J. Wattie ◽  
P. M. O’Byrne ◽  
E. E. Daniel
Keyword(s):  
Smooth Muscle ◽  
Electrical Field Stimulation ◽  
Tracheal Smooth Muscle ◽  
Organ Bath ◽  
Field Stimulation ◽  
Electrical Field ◽  
Concentration Response Curve

To investigate the role of prostaglandin (PG) E2 in allergen-induced hyperresponsiveness, dogs inhaled either the allergen Ascaris suum or vehicle (Sham). Twenty-four hours after inhalation, some animals exposed to allergen demonstrated an increased responsiveness to acetylcholine challenge in vivo (Hyp-Resp), whereas others did not (Non-Resp). Strips of tracheal smooth muscle, either epithelium intact or epithelium denuded, were suspended on stimulating electrodes, and a concentration-response curve to carbachol (10−9 to 10−5 M) was generated. Tissues received electrical field stimulation, and organ bath fluid was collected to determine PGE2content. With the epithelium present, all three groups contracted similarly to 10−5 M carbachol, whereas epithelium-denuded tissues from animals that inhaled allergen contracted more than tissues from Sham dogs. In response to electrical field stimulation, Hyp-Resp tissues contracted less than Sham tissues in the presence of epithelium and more than Sham tissues in the absence of epithelium. PGE2release in the muscle bath was greater in Non-Resp tissues than in Sham or Hyp-Resp tissues when the epithelium was present. Removal of the epithelium greatly inhibited PGE2release. We conclude that tracheal smooth muscle is hyperresponsive in vitro after in vivo allergen exposure only when the modulatory effect of the epithelium, largely through PGE2 release, is removed.

Activation of alpha-adrenergic response in tracheal smooth muscle: a postreceptor mechanism

1983 ◽  
Vol 54 (6) ◽  
pp. 1469-1476 ◽  
Author(s):  
P. J. Barnes ◽  
B. E. Skoogh ◽  
J. K. Brown ◽  
J. A. Nadel
Keyword(s):  
Smooth Muscle ◽  
Contractile Response ◽  
Electrical Field Stimulation ◽  
Tracheal Smooth Muscle ◽  
Adrenergic Blockade ◽  
Adrenergic Response ◽  
Receptor Number ◽  
Related Process

We have investigated the activation of alpha-adrenergic contractile responses in dog tracheal smooth muscle. After cholinergic and beta-adrenergic blockade, neither electrical field stimulation nor alpha-adrenergic agonists caused contraction of trachealis strips in vitro, but after exposure to histamine or serotonin a striking contractile response was obtained. Similar activation of the contractile response to norepinephrine was seen in isolated tracheal segments in vivo after exposure to histamine and serotonin. This response was mediated predominantly by alpha 2-adrenoceptors, because the alpha 2-antagonist yohimbine was a potent inhibitor whereas the alpha 1-antagonist prazosin was a weak inhibitor of the response to both electrical stimulation and exogenous agonists. Using [3H]yohimbine to label alpha 2-receptors and [3H]prazosin to label alpha 1-receptors, we confirmed the preponderance of alpha 2-receptors in trachealis membranes but found no increase in either receptor number or affinity after incubating muscle strips with histamine. The magnitude of alpha-adrenergic contraction was significantly related to the magnitude of precontraction by histamine and serotonin both in vitro and in vivo but persisted after washout. Acetylcholine was much less potent in activating the alpha-adrenergic response. We conclude that activation of airway alpha-adrenergic responses involves a postreceptor mechanism not directly related to membrane depolarization, but involving some related process such as activation of calcium channels.

Exposure of immature rats to hyperoxia increases tracheal smooth muscle stress generation in vitro

1994 ◽  
Vol 76 (2) ◽  
pp. 743-749 ◽  
Author(s):  
M. B. Hershenson ◽  
M. E. Wylam ◽  
N. Punjabi ◽  
J. G. Umans ◽  
P. T. Schumacker ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Contractile Function ◽  
Stress Generation ◽  
Tracheal Smooth Muscle ◽  
Smooth Muscle Contractility ◽  
Cross Sectional ◽  
Induced Stress ◽  
Hyperoxic Exposure

Recently, we demonstrated that chronic exposure to hyperoxia causes in vivo airway muscarinic receptor hyperresponsiveness in the developing rat [Am. J. Physiol. 262 (Lung Cell. Mol. Physiol. 6): L263-L269, 1992]. To test whether airway cholinergic hyperresponsiveness might result from intrinsic alterations in smooth muscle contractility, we measured the effect of in vivo hyperoxia on the contractile force elicited by acetylcholine (ACh) of isometrically mounted tracheal rings in vitro. Tracheal rings were obtained from 3-wk-old rats exposed to air or to > 95% O2 for 8 days. Muscarinic responses were determined by measuring the force elicited by exposure to increasing concentrations of ACh. Responses were normalized to the morphometrically determined tracheal smooth muscle cross-sectional area in a plane perpendicular to the axis of force generation. In vivo O2 exposure significantly increased maximal ACh-induced stress generation (response to 10(-3) M ACh: air, 15.92 +/- 1.37 g/mm2; O2, 21.78 +/- 1.52 g/mm2; P = 0.010). The ACh-induced stress generation of cylinders from hyperoxic rats was substantially reduced by both epithelial removal and treatment with the cyclooxygenase inhibitor indomethacin. We conclude that in vivo hyperoxic exposure increases tracheal smooth muscle contractile function in vitro and that epithelium-derived prostaglandin(s) contributes to the observed increase in maximal contractile responsiveness.

Increased in vitro responses of tracheal smooth muscle from hyperresponsive guinea pigs

1990 ◽  
Vol 68 (4) ◽  
pp. 1316-1320 ◽  
Author(s):  
K. Ishida ◽  
P. D. Pare ◽  
R. J. Thomson ◽  
R. R. Schellenberg
Keyword(s):  
Smooth Muscle ◽  
Guinea Pigs ◽  
Airway Hyperresponsiveness ◽  
Contractile Activity ◽  
Tracheobronchial Tree ◽  
Force Generation ◽  
Tracheal Smooth Muscle ◽  
Antigen Challenge

Repeated aerosol antigen challenge of previously sensitized guinea pigs induces airway hyperresponsiveness to inhaled acetylcholine. To determine the mechanism producing these airway changes and assuming that changes in the trachealis muscle reflect changes in muscle of the entire tracheobronchial tree, we examined the in vitro smooth muscle mechanics and morphometric parameters of tracheae from guinea pigs demonstrating hyperresponsiveness in vivo vs. tracheae from control guinea pigs. No differences between these groups were found in luminal volume at zero transmural pressure, passive pressure-volume characteristics, or area of airway wall. Smooth muscle areas were slightly less in tracheae from hyperresponsive guinea pigs. Tracheae from hyperresponsive guinea pigs had both significantly increased isovolumetric force generation and isobaric shortening compared with tracheae from controls when evaluated over the range of transmural pressures from -40 to 40 cmH2O. We conclude that the in vivo airway hyperresponsiveness induced with repeated antigen challenge is associated with both increased force generation and shortening of tracheal smooth muscle without increased muscle mass, suggesting enhanced contractile activity.

Increased in vitro airway responsiveness in sheep following repeated exposure to antigen in vivo

1985 ◽  
Vol 59 (6) ◽  
pp. 1874-1878 ◽  
Author(s):  
E. M. Wagner ◽  
S. R. Kleeberger ◽  
E. W. Spannhake ◽  
G. K. Adams
Keyword(s):  
Smooth Muscle ◽  
Thromboxane A2 ◽  
Airway Responsiveness ◽  
Tracheal Smooth Muscle ◽  
Prostaglandin F2 Alpha ◽  
Peripheral Airway ◽  
Exposure In Vivo ◽  
Thromboxane A2 Analogue

We have studied the effect of repeated in vivo antigen exposure on in vitro airway responsiveness in sensitized sheep. Fourteen sheep underwent five biweekly exposures to aerosolized Ascaris suum antigen or saline. Following this exposure regimen, the animals were killed and tracheal smooth muscle and lung parenchymal strips were prepared for in vitro studies of isometric contraction in response to histamine, methacholine, prostaglandin F2 alpha, and a thromboxane A2 analogue. No alteration in tracheal smooth muscle responsiveness was observed between saline- and antigen-exposed tissue. In contrast, by use of lung parenchymal strips as an index of peripheral airway responsiveness, significant increases in responsiveness to histamine and a thromboxane A2 analogue (10(-6) and 10(-5) M) were observed in antigen-exposed tissue compared with saline controls. These results demonstrate that repeated antigen exposure in vivo selectively increase the responsiveness of peripheral lung smooth muscle to certain chemical mediators of anaphylaxis.

Responsiveness of isolated tracheal smooth muscle from normal and sensitized guinea pigs

1987 ◽  
Vol 65 (9) ◽  
pp. 1942-1950 ◽  
Author(s):  
S. Mansour ◽  
E. E. Daniel
Keyword(s):  
Smooth Muscle ◽  
Tracheal Smooth Muscle ◽  
Field Stimulation ◽  
In Vitro Techniques ◽  
Total Magnitude ◽  
Specific Receptors ◽  
Relaxation Responses ◽  
Selective Change

Studies of the responsiveness of strips of tracheal smooth muscle and the changes after sensitization of ovalbumin were carried out. The hypothesis that there might be a generalized or a selective change of airway smooth muscle responsiveness to sensitization was examined in vitro. Agonists acting on muscarinic receptors, α1-, α2-, and β-adrenoceptors, purine receptors, histamine and serotonin receptors, and leukotriene and prostaglandin receptors were tested, as well as mediators released from local nerves by field stimulation and procedures such as elevation of potassium or addition of Ca2+ ionophores which do not involve specific receptors. Sensitivity to serotonin increased significantly in sensitized animals. Total magnitude of the contraction and subsequent relaxation responses to field stimulation also increased significantly. Neither of these changes was large in magnitude. Although there were a few minor changes in sensitivity (pD2) or in maximum responses, the hypothesis of important changes in responses of any sort in tracheal muscle after sensitization was rejected. The question was raised whether this general absence of changed responsiveness in vitro reflected (i) the failure of sensitization to induce generalized smooth muscle hyperresponsiveness, (ii) the loss of the mechanisms of such responsiveness in vitro, or (iii) the inadequacy of in vitro techniques to assess responsiveness present in vivo.

Phosphorylation of calponin in airway smooth muscle

1997 ◽  
Vol 272 (1) ◽  
pp. L115-L123 ◽  
Author(s):  
J. Pohl ◽  
S. J. Winder ◽  
B. G. Allen ◽  
M. P. Walsh ◽  
J. R. Sellers ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Inhibitory Effect ◽  
Tracheal Smooth Muscle ◽  
Actin Binding ◽  
In Vitro Motility Assay ◽  
In Vitro Motility ◽  
Kinase C ◽  
Gradient Gel Electrophoresis

Calponin is an actin-binding protein known to be a substrate in vitro for several protein kinases and phosphoprotein phosphatases. We tested the hypothesis that calponin is phosphorylated in vivo using canine tracheal smooth muscle strips metabolically labeled with 32Pi. Calponin was gel purified from muscles stimulated with 1 microM carbachol. Phosphorylation increased to 2.0 times the basal level of 178 +/- 26 counts per minute (cpm)/microgram calponin within 30 s to 350 +/- 64 cpm/micrograms. Two-dimensional nonequilibrium pH gradient gel electrophoresis resolved four charge isoforms of calponin in unstimulated muscle. Stimulation with carbachol induced an additional more acidic isoform. Phosphorylation of calponin in vitro with protein kinase C (PKC) also induced formation of additional acidic isoforms. The functional effect of phosphorylation was demonstrated using an in vitro motility assay in which unphosphorylated calponin (2 microM) caused a profound inhibition of actin sliding. Calponin phosphorylated by PKC did not inhibit actin sliding. The results show that phosphorylation of calponin occurs in intact tracheal smooth muscle and that phosphorylation of calponin in vitro alleviates the inhibitory effect of calponin on actomyosin function.

scholarly journals Efficacy of fexofenadine in isolated rat tracheas

2013 ◽  
Vol 51 (4) ◽  
pp. 376-380
Author(s):  
W.S. Lai ◽  
Y.Y. Lin ◽  
Y.H. Chu ◽  
C.H. Wang ◽  
H.W. Wang
Keyword(s):  
Smooth Muscle ◽  
Electrical Field Stimulation ◽  
Histamine Receptor ◽  
Tracheal Smooth Muscle ◽  
Dependent Manner ◽  
Chemical Mediator ◽  
Bronchial Inflammation ◽  
Isolated Rat

Objectives: Histamine is an important chemical mediator in both nasal and bronchial inflammation in patients with allergic rhinitis and asthma. The effect of histamine receptor-1 antagonists on nasal mucosa in vivo is well known, however, the effect on tracheal smooth muscle has rarely been explored. The purpose of this study was to determine the effects of fexofenadine on isolated tracheal smooth muscle in vitro. Methods: Six tracheal strips were used for each experiment, and one untreated strip served as a control. We examined the effectiveness of fexofenadine on isolated rat tracheal smooth muscle by testing the effect on: 1) tracheal smooth muscle resting tension; 2) contraction caused by 10E-6 M methacholine as a parasympathetic mimetic; and 3) electrically induced tracheal smooth muscle contractions. Results: The results indicated that addition of methacholine caused the trachea to contract in a dose-dependent manner. The addition of fexofenadine at a dose of 10E-4 M elicited a significant relaxation response compared to 10E-6 M methacholine-induced contraction. There were no detectable changes in the peak tension of electrical field stimulation-induced contractions in the fexofenadine group. Conclusion: High concentrations of fexofenadine had an anti-cholinergic effect. In addition to diminishing histamine-mediated allergic symptoms, fexofenadine may have a potentially therapeutic implication in alleviating asthma-related symptoms due to reducing methacholine-induced contractions of tracheal smooth muscle though these aspects were not studied.

Sign in / Sign up

Export Citation Format

Share Document