Effects of cholera toxin on vasoconstriction and cyclic AMP content of the isolated rabbit ear artery

We have studied the effect of cholera toxin on the constrictor responses of the isolated, perfused rabbit ear artery to nerve stimulation and to norepinephrine infusion. We found that when we perfussed arteries with cholera toxin (1–9 μg/ml) for five minutes or longer, the toxin gradually inhibited the responses to intermittent stimulation of the adrenergic nerves and to brief infusion of norepinephrine. The constrictor responses began to decrease between one and two hours after we added cholera toxin, and the responses were still depressed after 24 hours. Cholera toxin inhibited both the rapid, initial phase and the slower, sustained phase of the biphasic response of the ear artery to nerve stimulation. Propranolol and indomethacin did not block the effect of cholera toxin on vasoconstriction. However, when we mixed the toxin with antitoxin or G_M1 ganglioside, we prevented the inhibitory effect on vasoconstriction. Levels of adenosine 3′:5′-cyclic monophosphate (cyclic AMP) in arteries treated with cholera toxin were greater than levels of cyclic AMP in untreated arteries. The cyclic AMP content increased and the constrictor responses decreased with a similar time course after the arteries were exposed to the toxin. Thus an increase in cyclic AMP may be involved in the relaxation of vascular smooth muscle induced by cholera toxin.     

Seasonal variation of cold-induced vasooscillation on rabbit ear central artery

We studied the seasonal variation of vasooscillation of a rabbit ear central artery induced by exposure of the earlobes to - 7 degrees C liquid. The data were collected over a period of 10 years and analyzed by month. a) The index of arterial temperature fluctuation (IATF), i.e., activation index of cold-induced vasooscillation (CIVO), ranged from 114.5 +/- 26.7 (mean +/- SE) in January to 386.7 +/- 36.1 in June. A significant variation over all 12 months was revealed by analysis of variance (P less than 0.01). The values measured in May (317.1 +/- 47.3), June (386.7 +/- 36.1), and July (315.1 +/- 36.0) were significantly larger than those of other months. b) The monthly IATFs were correlated with the open air temperatures (r = 0.7017, P less than 0.05); however, the peak IATF occurred in June, while the peak open air temperature was in August. c) There was no seasonal variation of the arterial temperature either before or at 18-20 min after -7 degrees C immersion. Arterial temperature was not related to IATF during -7 degrees C exposure. d) When measuring-site temperature was steady, the thermistor temperature changed in parallel with the output from a laser blood volume meter. e) The CIVO was independent of systemic blood pressure and heart rate, which suggested that the occurrence of CIVO was regulated by changes in local vascular resistance.     

Electro- and pharmacomechanical coupling in the smooth muscle cells of the rabbit ear artery

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.     

Sodium and calcium interactions in vascular smooth muscle cells of the rabbit ear artery

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.