Ouabain-induced Contraction Research Articles

Mechanism of extraneuronal adrenaline release induced by K+ -free medium and ouabain, but not by high KCl in vas deferens of guinea-pig and rat

The effects of K+-free medium, ouabain or high KCl on the contractile response and spontaneous efflux of adrenaline were analyzed in denervated guinea-pig or rat vasa deferentia preloaded with 3 x 10(-5) M (+/-)-adrenaline. The K+-free medium-induced contraction of guinea-pig tissues was related to the concentration of adrenaline used during preloading and was suppressed by the addition of 2.5 mM Rb+ but not by 2.5 mM Cs+. The contractile effects of K+-free medium and 10(-5) M ouabain, but not the response to high KCl, were prevented by 5 x 10(-6) M phentolamine or 3 x 10(-5) M deoxycorticosterone. These contractions, therefore, appear to be mediated by adrenaline released from the extraneuronal compartment. The K+-free medium- or ouabain-induced contractions of denervated rat vasa deferentia were small in comparison with those of the guinea-pig tissue. The amount of spontaneous efflux of adrenaline from the guinea-pig preparation was significantly enhanced by omission of K+ or by 10(-4) M ouabain but not by 80 mM KCl. K+-free medium and 3 x 10(-5) M ouabain merely elicited a small membrane depolarization accompanied by a slight decrease of membrane resistance in the guinea-pig vas deferens, whereas 30 mM KCl markedly altered these electrical parameters. From these findings, it is suggested that the rapid increase in extraneuronal amine release following inhibition of Na+-K+-ATPase may be due to some direct effect of K+ deficiency or of ouabain on the binding between the enzyme and the catecholamine rather than to alterations of membrane properties as a consequence of enzyme inhibition.

Mechanisms of ouabain-induced arterial muscle contraction.

Ouabain caused dose-related contractions in helically cut strips of cerebral, mesenteric, renal, and femoral arteries isolated from dogs and monkeys; the contractions were elicited at lower concentrations in cerebral than in the peripheral arteries. Contractions induced by ouabain were abolished by exposure of cerebral arteries to Ca2+-free media. Contractions induced by Ca2+ in Ca2+-free media were potentiated by ouabain. Substitution of LiCl for NaCl and reduction of [K+]0 suppressed the contractile response of cerebral arteries to ouabain. Pretreatment of dogs with reserpine abolished the response of peripheral arteries to ouabain but not the cerebroarterial response. Phentolamine also abolished the response of peripheral arteries. Once oubain-induced contractions were stabilized, the alpha-antagonist caused a marked relaxation in peripheral but not in cerebral arteries. Ouabain-induced contractions appear to be mediated by an increase in the Ca2+ influx that results mainly from an inhibition of the electrogenic Na+ pump in low concentrations (cerebral arteries) and from a release of norepinephrine from nerves in high concentrations (mesenteric, renal, and femoral arteries).

Mechanisms of action of transmitters and other substances on smooth muscle.

Mechanisms of action of transmitters and other substances on smooth muscle.

Neuronal norepinephrine as mediator for ouabain-induced smooth muscle contraction

With some latency, ouabain (10 −6-10 −5 M) induced a long-lasting contractile response of the isolated vas deferens of the guinea-pig. The ouabain-induced contraction was potentiated by pretreatment with tropolone, whereas it was prevented by pretreatment with cocaine, bretylium, reserpine or phentolamine. The response was unaffected by atropine, methysergide and tetrodotoxin. Ouabain, perfused into the isolated tissue, enhanced dose-dependently the release of norepinephrine into the medium. However, this enhancement of norepinephrine release was prevented by reserpine or by removal of Ca from the medium. These findings support the hypothesis that the ouabain-induced contraction of vas deferens might be due to norepinephrine released from the adrenergic nerve ending granules through a Ca 2+-dependent process.