Influence of some variables in the Triton X-100 method of skinning the plasmalemmal membrane from guinea pig trachealis muscle

Cortijo, Julio; Dixon, J.S.; Foster, Robert W.; Small, R.C.

doi:10.1016/0160-5402(87)90075-1

Cited by 22 publications

(10 citation statements)

References 15 publications

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“…Using this technique we found it was unnecessary to add exogenous calmodulin in order to generate a contraction to Ca2". In this regard we confirm the observations of Sparrow et al (1984) and Cortijo et al (1987). However, we found that addition of calmodulin (1 tmoll-') failed to potentiate the Ca2+-induced contractions in our skinned trachealis preparations (data not shown).-This finding is contrary to that observed by Sparrow et al (1984) but is in complete agreement with the recent observations of Cortijo et al (1987).…”

Section: Discussionsupporting

confidence: 92%

“…This conclusion is supported by the finding that calmidazolium, a known inhibitor of calmodulin (Gietzen et al, 1981) Sparrow et al (1984). Since the plasma membrane is markedly disrupted by the skinning treatment (Cortijo et al, 1987), these effects of cyclic AMP and its kinase cannot be ascribed to either a stimulant effect upon ATP-dependent Ca2" extrusion mechanisms or an inhibitory effect upon the influx of extracellular Ca2" through membrane calcium channels. In other smooth muscles Triton X-l00 is found to destroy the functional integrity ofboth the sarcoplasmic reticulum and the mitochondria (Endo et al, 1982;Stout & Diecke, 1983).…”

Section: Discussionmentioning

confidence: 90%

“…SKF 94120 is a recently described selective inhibitor ofthe 'low Ki' phosphodiesterase enzyme (PDE III) in guinea-pig and human tissues (Gristwood et al, 1985;Reeves et al, 1987 be manipulated directly and the effects on the contractile machinery of the cell to be studied without the intervening influence of the plasma membrane and, under certain conditions, the sarcoplasmic reticulum. Several different methods for chemically skinning airway smooth muscle have been described recently (Ito & Itoh, 1984;Sparrow et al, 1984;Hashimoto et al, 1985;Allen et al, 1986a, b;Cortijo et al, 1987).…”

Section: Discussionmentioning

confidence: 99%

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Effects of phosphodiesterase inhibitors on normal and chemically‐skinned isolated airway smooth muscle

Bryson¹,

Rodger²

1987

British J Pharmacology

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1 The effects of three phosphodiesterase inhibitors (papaverine, igobutyl methyl xanthine (IBMX) and SKF 94120) were examined on tension responses and cyclic nucleotide content (both cyclic AMP and cyclic GMP) of normal and Triton X-100 skinned isolated trachealis of the guinea-pig. 2 The three inhibitors were approximately equipotent in eliciting concentration-dependent relaxation of histamine-induced contractions of the trachealis. 3 Papaverine-induced relaxation was associated with concentration-related increases in the levels of both cyclic nucleotides. 4 IBMX at low concentrations (1 gmol 1-') produced significant relaxation (36%) of histaminecontracted trachealis without changing cyclic nucleotide levels. At a ten fold higher concentration IBMX-induced relaxation (95%) was associated with a selective increase in tissue cyclic GMP levels.Only at the highest concentration tested (100 gmol I-) did IBMX increase cyclic AMP levels significantly.5 SKF 94120(1 pmol 1-') elicited a 23% relaxation of the contracted trachealis without altering the tissue content ofeither cyclic nucleotide. At the two higher concentrations tested (10 and 100 pmol 1 -'), SKF 94120-induced relaxation was accompanied by a selective increase in the levels of cyclic AMP. 6 In the skinned trachealis Ca2" (10 and 20 gmol I ')-induced contractions were significantly inhibited by the calmodulin antagonist calmidazolium (10 gtmol 1I-') and by cyclic AMP (10 pmol 1 1'), the catalytic subunit of cyclic AMP-dependent protein kinase (0.1 pmol I') and cyclic GMP (10 pmol l'). 7 Papaverine (1O00Imoll-') significantly inhibited (31 ± 6%) the Ca2t-induced contractions of the skinned trachealis. Both IBMX and SKF 94120 were without effect. 8 It is concluded that cyclic nucleotide-dependent mechanisms have an inhibitory action on the biochemical processes that lead to contraction of the guinea-pig trachealis. The results suggest that a functional sarcoplasmic reticular and/or plasma membrane is essential for the expression of IBMXand SKF 94120-induced relaxation. This is not the case for papaverine. The results also highlight the fact that significant relaxant responses of airway smooth muscle can be produced by phosphodiesterase-inhibiting drugs without concomitant elevations in tissue cyclic nucleotide content.

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Section: Discussionsupporting

confidence: 92%

Section: Discussionmentioning

confidence: 90%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Effects of phosphodiesterase inhibitors on normal and chemically‐skinned isolated airway smooth muscle

Bryson¹,

Rodger²

1987

British J Pharmacology

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“…Using skinned trachea preparations, a possible mod ification of the contractile apparatus per se due to sensiti zation was investigated by Cortijo et al [16]; no difference could be found between sensitized and control guinea pigs.…”

Section: Discussionmentioning

confidence: 99%

Enhancement of the Responsiveness of Vas Deferens and Ileum Smooth Muscle in Sensitized Guinea Pigs: In vitro Study

Bidon¹,

Blin²,

Gogny³

et al. 1994

Int Arch Allergy Immunol

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Changes in the reactivity of the ileum (to histamine and barium chloride) and vas deferens (to acetylchohne and barium chloride), isolated from actively egg albumen-sensitized guinea pigs, have been investigated. The study was performed on 2 guinea pig strains: the Dunkin-Hartley strain, usually used as an airway allergic model, and the BFA strain. In actively sensitized guinea pigs of both strains, concentration-response curves exhibited a significant dose-dependent upward shift compared to those obtained in control guinea pigs. The maximal contraction strength calculated from these curves was significantly enhanced in both sensitized guinea pig strains, without a change in EC₅₀ values. This study showed that the active antigen sensitization procedure involved several smooth muscle functions, and not exclusively the trachea.

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“…These possible loci of antispasmogenic action were examined by studying the effects of BHA, BHT and nifedipine on skinned fibres. The skinning procedure we adopted, based on Triton X-100, not only disrupts the plasma membrane (Cortijo et al, 1987) but has also been found to destroy the functional integrity of both the sarcoplasmic reticulum and the mitochondria (Meisheri & Riiegg, 1983).…”

Section: Discussionmentioning

confidence: 99%

Calcium antagonist and antiperoxidant properties of some hindered phenols

Sgaragli

Valoti

Gorelli

et al. 1993

British J Pharmacology

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1 The calcium antagonist and antioxidant activities of certain synthetic and natural phenols, related to BHA (2-t-butyl-4-methoxyphenol), were evaluated in rat ileal longitudinal muscle and in lipid peroxidation models respectively. 2 Compounds with a phenol or a phenol derivative moiety, with the exception of 2,2'-dihydroxy-3,-3'-di-t-butyl-5,5'-dimethoxydiphenyl (di-BHA), inhibited in a concentration-dependent manner the BaCl2-induced contraction of muscle incubated in a Ca2+-free medium. Calculated pIC50 (M) values ranged between 3.32 (probucol) and 4.96 [3,5-di-t-butyl-4-hydroxyanisole (di-t-BHA)], with intermediate activity shown by khellin

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Influence of some variables in the Triton X-100 method of skinning the plasmalemmal membrane from guinea pig trachealis muscle

Cited by 22 publications

References 15 publications

Effects of phosphodiesterase inhibitors on normal and chemically‐skinned isolated airway smooth muscle

Effects of phosphodiesterase inhibitors on normal and chemically‐skinned isolated airway smooth muscle

Enhancement of the Responsiveness of Vas Deferens and Ileum Smooth Muscle in Sensitized Guinea Pigs: In vitro Study

Calcium antagonist and antiperoxidant properties of some hindered phenols

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