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  • 1
    ISSN: 1432-1912
    Keywords: Isolated trachea ; Preganglionic stimulation ; Transmural stimulation ; [3H]Acetylcholine release ; [3H]Phosphorylcholine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Basal and stimulated outflow of radioactive acetylcholine, phosphorylcholine and choline from rat and guinea-pig isolated tracheae were measured by reverse phase HPLC followed by liquid-scintillation-spectrometry. Tracheae were stimulated either by an electrical field (transmural stimulation) or by a local stimulation of the innervating parasympathetic nerves (preganglionic stimulation). Epithelium was removed in most experiments, as the epithelium inhibits acetylcholine release. The basal tritium efflux (1,600 dpm/3min) from rat isolated tracheae incubated with [3H]choline consisted of 56% [3H]phosphorylcholine and 38% [3H]choline. Preganglionic stimulation (15 Hz, 1,200 pulses) caused a 2-fold increase in tritium outflow that was abolished by the removal of extracellular calcium or by the addition of tetrodotoxin. The stimulated outflow of tritium induced by preganglionic nerve stimulation was caused by an exclusive release of [3H]acetylcholine, whereas the efflux of [3H]phosphorylcholine and [3H]choline remained unaffected by this stimulation mode. Transmural stimulation of the rat or guinea-pig trachea, however, caused, in addition to the release of [3H]acetylcholine, the outflow of [3H]phosphorylcholine. Hexamethonium (300 μmol/l) or tubocurarine (100 μmol/l) inhibited (80%) the increase in tritium outflow evoked by preganglionic stimulation, but did not affect tritium outflow evoked by transmural stimulation. Oxotremorine reduced [3H]acetylcholine release evoked by both stimulation modes, but oxotremorine was less potent with transmural stimulation. Scopolamine (0.3 μmol/l) enhanced (120%) the release of [3H]acetylcholine evoked by preganglionic nerve stimulation indicating the blockade of an endogenous negative muscarinic feedback mechanism. Epithelium-dependent inhibition of [3H]acetylcholine release was evident with both preganglionic and transmural stimulation. The present experiments demonstrate the release of [3H]acetylcholine evoked from the isolated trachea by stimulation of the preganglionic trunk of the parasympathetic cholinergic nerves. Qualitative and quantitative differences were observed in comparison to transmural stimulation. Preganglionic nerve stimulation allows a selective excitation of pulmonary, parasympathetic nerve fibres, mimics the physiological excitation of intramural neurones and is not followed by the liberation of phosphorylcholine from non-neuronal cells.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-1912
    Keywords: Longitudinally opened trachea ; Tube-preparation ; Cromakalim ; Epithelium ; Inhibition of [3H]acetylcholine release
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Rat isolated tracheae were labelled by incubation with [3H]choline to measure the tritium efflux elicited by electrical stimulation of the extrinsic parasympathetic nerves in vitro. Stimulated tritium efflux reflects the neuronal release of newly synthesized acetylcholine; the effects of potassium channel openers on the stimulated tritium efflux were investigated. In tracheae opened longitudinally neither cromakalim nor its 3S,4R-enantiomer, BRL 38227, reduced the stimulated tritium efflux, whereas in intact tube-preparations cromakalim (0.01–1 μmol/l) mediated a concentration-dependent inhibition. The inhibitory effect of 1 μmol/l cromakalim was prevented by 0.1 μmol/l glibenclamide. Likewise, BRL 38227 (0.01 and 0.1 μmol/l) inhibited the stimulated tritium efflux, but the inhibitory effect vanished at high concentrations (1 and 10 μmol/l). The 3R,4S-enantiomer of cromakalim, BRL 38226 (0.1, 1 and 10 μmol/l), on its own did not significantly inhibit the stimulated tritium efflux, but a combination of both enantiomers (0.5 or 1 μmol/l of each) produced an inhibition similar to that caused by 1 μmol/l cromakalim. In epithelium-denuded tube-preparations neither cromakalim nor BRL 38227 reduced the stimulated tritium efflux. The mucosal/submucosal microenvironment is better preserved in intact tube-preparations than in longitudinally-opened tracheae which are cut along their whole length so that the luminal surface is exposed directly to the surrounding medium. The present experiments show an neuronal inhibitory effect of cromakalim which is mediated by an epithelium-dependent mechanism.
    Type of Medium: Electronic Resource
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