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  • Title: Cytosolic calcium concentration-force relation during contractions in the rabbit femoral artery: time-dependency and stimulus specificity.
    Author: Fukuizumi Y, Kobayashi S, Nishimura J, Kanaide H.
    Journal: Br J Pharmacol; 1995 Jan; 114(2):329-38. PubMed ID: 7881732.
    Abstract:
    1. By use of front-surface fluorometry with fura-2-loaded rabbit femoral arterial strips, both the cytosolic Ca2+ concentration ([Ca2+]i) and force were simultaneously monitored. By utilizing the [Ca2+]i-force curves, we were thus able to examine the temporal changes in the relationships between [Ca2+]i and force ([Ca2+]i-force relationship) during contractions induced by a high external K+ solution, noradrenaline (NA) and 5-hydroxytryptamine (5-HT). 2. The 'basic' [Ca2+]i-force relationship of the Ca(2+)-induced contractions was obtained by the cumulative applications of extracellular Ca2+ (0-10 mM) during 118 mM K(+)-depolarization (Ca(2+)-contractions). 3. When each vascular strip was exposed to high external K+ (30 mM K(+)-118 mM K+) solutions, the [Ca2+]i abruptly increased until it reached a peak, and then slightly decreased and eventually reached a steady-state level. The force also rapidly rose to reach a maximum plateau level. The changes in [Ca2+]i were more rapid than those in the force. Thus, the [Ca2+]i-force curves observed during the contractions induced by high+ (30 mM-118 mM) solutions showed a counter-clockwise rotation, over time. The entire curve shifted to the right, in a concentration-dependent manner, as compared with the line of the 'basic' [Ca2+]i-force relationship of the Ca(2+)-contraction. However, the [Ca2+]i-force relationship of the steady-state of contractions induced by the single dose applications of high K+ (30 mM-118 mM) overlapped with the line of the 'basic' [Ca2+]i-force relationship of Ca(2+)-contractions. 4. As references, the levels of [Ca2+]i and the force at rest (without stimulation) and at the steady-state of the contractions induced by a single dose application of 118 mM K+ solution were designated as 0% and 100%, respectively. When the vascular strips were exposed to NA (10(-5) M) and to 5-HT (10(-4) M), the [Ca2+]i abruptly rose, and reached a peak (107.1 +/- 5.8%) and 101.3 +/- 2.8%, respectively) after 1 min and 2 min, respectively (the [Ca2+]i-rising phase), and thereafter declined with a similar time course (the [Ca2+]i-declining phase) until reaching a low steady level (the steady-state phase). The force induced by 10(-5) M NA and 10(-4) M 5-HT reached a peak at 4 min (129%) and at 2 min (115%), respectively, and thereafter gradually declined. In contrast to the similarity in the [Ca2+]i transient between NA and 5-HT, the force induced by NA declined more slowly and reached higher steady levels than that seen with 5-HT. The level of force 20 min after the application of NA and 5-HT was 112% and 72%, respectively.5. In the entire time course of the 5-HT-induced contraction, i.e., in [Ca2+]i-rising, [Ca2+]i-declining and the steady-state phases, the [Ca2+]i-force relation was almost the same as that of the Ca2+-contractions.In the [Ca2+]i-rising phase of NA-induced contraction, the [Ca2+]i-force relation was similar to that of the Ca2+-contractions. However, in the [Ca2+] -declining and the steady-state phases, NA produced a greater force than that expected from a given change in the [Ca2+]i of the Ca2+-contractions, which resulted in a leftward shift of the [Ca2+]i-force relation. The extent of the leftward shift depended on the concentration of NA.6. These results suggest that (1) changes in [Ca2+]i precede changes in the force during the high K+-induced contraction, (2) in the initial [Ca2+]i-rising phase of the contractions induced by NA or by 5-HT, the [Ca2+]i-force relation is similar to that of Ca2+-contractions, and (3) in the subsequent[Ca2+]i-declining and the steady-state phases of the contractions, 5-HT demonstrated little enhancement in force for the given levels of [Ca2+]i, while NA induced a greater force for the given levels of [Ca2+],, in the rabbit femoral artery. Based on the above findings we suggest the presence of a time-dependent and stimulus-specific modulation of the Ca2+ sensitivity in the contractile apparatus of arterial smooth muscles.
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