The effects of Na-free and of K-free solutions on the membrane potential, on tension development, and on 45Ca exchange have been investigated in rabbit ear artery. The contraction induced by Na-free solutions and the tension which develops in K-free solutions after a delay of about 1 h are both submaximal. Exposure for 4 h to K-free solutions does not affect the membrane potential, whereas Na-free solutions depolarize the cells by 10-20 mV, depending on the Na-substitute. Neither the amplitude nor the rate constant of the slowly exchanging 45Ca-fraction is affected by these experimental procedures. Substituting external Na by choline or TMA induces a transient increase of the 45Ca-efflux rate which does not occur in a Ca-free efflux medium, and which can be blocked with La. K readmission to Na-enriched tissues hyperpolarizes the cells up to -100 mV and induces a relaxation, without exerting any effect on the 45Ca efflux rate. The release of Ca from intracellular stores, induced by histamine and FCCP, and its subsequent extrusion through the plasma membrane produce a transient stimulation of the 45Ca efflux, which is not affected by the reduction of the Na gradient. The transient contraction induced by histamine in Ca-free solutions is affected in a different way by different Na substitutes. The results do not fit the Na-Ca exchange hypothesis but are consistent with an effect of the Na gradient on the passive Ca influx.

A contraction of the rabbit ear artery can be induced by depolarizing the cells with a K-rich solution if Ca is present. 10(-9)-10(-6) M noradrenaline and 10(-8)-10(-7) M histamine cause a contraction of this tissue without modifying the membrane potential. If the histamine concentration exceeds 10(-7) M some depolarization of the membrane also occurs. Both noradrenaline and histamine also induce a contraction in Ca-free medium, even if La is present. None of these stimuli produces action potentials or fluctuations of the membrane potential. Besides these tonic contractions, the ear artery can also produce phasic contractions when 10 mM TEA is added to the medium. Such contractions are caused by the appearance of action potentials which are Ca dependent and which are similar to those appearing in visceral smooth muscle. A study of 45Ca fluxes has revealed that K depolarization and noradrenaline cause only a small increase in 45Ca uptake by the cells, while noradrenaline also releases cellular Ca, even in Ca-free medium. A comparison of tension development and 45Ca release induced by noradrenaline in Ca-free medium suggests that Ca extrusion could be very efficient in the rabbit ear artery and that it could play a direct role in its relaxation.

The distribution of Ca in the cellular compartment of smooth muscle cells of the rabbit aorta has been studied by analyzing the effect of norepinephrine, caffeine, and DNP on 45Ca exchange and on the pattern of tension development. These three substances increase the release of 45Ca from the tissue, but DNP acts more slowly than norepinephrine or caffeine. Also, the effect of norepinephrine and caffeine on tension development occurs almost immediately, while that of DNP appears only after a delay of 5 min. Study of the effect of these substances on the Ca efflux has shown that norepinephrine and caffeine act probably on the same Ca compartment, while DNP seems to act on a different compartment with a slower exchange rate. The difference between these two pools could be further demonstrated by studying Ca release after loading the tissues with tracer in either K-rich solution or in a solution with reduced [Ca]o. The K depolarization results in an excessive loading of the cells with 45Ca. Exposing these cells during the efflux procedure to a solution containing DNP causes a much larger release of 45Ca than that observed after a loading procedure in normal solution. In contrast, the release of 45Ca elicited in such tissues by norepinephrine or caffeine disappears. This disappearance is due to the prolonged increase of the Ca exchangeability induced by K depolarization. During initial exposure to PSS the increased exchangeability causes an accelerated loss of tracer from the tissue compartment on which norepinephrine and caffeine act, while the DNP sensitive compartment is not affected. It is suggested that noradrenaline and caffeine act on the same calcium pool close to the membrane and that DNP acts mainly on the mitochondria.