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Department of Physiology, University of Nevada School of Medicine, Reno 89557-0046.
1. The relationships between cytosolic Ca2+ ([Ca2+]cyt; expressed as a fluorescence ratio at 400 nm and 500 nm using Indo-1) and contractile force was examined in strips of circular smooth muscles of canine gastric antrum. Rhythmic increases in [Ca2+]cyt were observed and contractions were biphasic. 2. In most muscles (70%), the amplitude of the second phase of the Ca2+ transient was less than or equal to the first phase of the Ca2+ transient, but the second phase of the contraction was much smaller than the first phase, suggesting a decrease in Ca2+ sensitivity during the second contractile phase. In 30% of muscles, the amplitude of the second phase of the Ca2+ transient was 2- to 3-fold greater than the first phase. In these muscles, the second phase of contraction was 10-fold greater than the first phase of contraction. Thus, a non-linear relationship between [Ca2+]cyt and force greatly amplifies force development when [Ca2+]cyt exceeds a threshold level. 3. Acetylcholine (ACh, 0.3-1 microM) increased the amplitudes of Ca2+ transients and basal [Ca2+]cyt between phasic contractions. The increase in basal [Ca2+]cyt did not cause tone to develop. ACh increased the amplitude of Ca2+ transients 2- to 3-fold and this was associated with a 15 to 20-fold increase in the force of phasic contractions. Pentagastrin (0.5 nM) and cholecystokinin octapeptide (CCK, 40 nM) had similar effects on Ca2+ transients and phasic contractions. 4. Bay K 8644 (0.1 microM) and TEA (5 mM) also increased the amplitudes of Ca2+ transients by 2- to 3-fold and phasic contractions by 15- to 30-fold. There was no significant difference observed between the [Ca2+]cyt-force relationships in the presence of agonists (i.e. ACh, pentagastrin and CCK) or when [Ca2+]cyt was increased by Bay K 8644 or TEA. These data suggest that agonist-dependent increases in Ca2+ sensitivity may not significantly regulate the [Ca2+]cyt-force relationship in antral muscles. 5. D600 (5 microM), added during stimulation with ACh (0.3 M), decreased [Ca2+]cyt and force without affecting the [Ca2+]cyt-force relationship. 6. Mechanisms exist for agonist-mediated enhancement of the Ca(2+)-force relationship. In alpha-toxin-permeabilized antrum, ACh (10 microM) with GTP (100 microM) or GTP gamma S (100 microM) increased the Ca(2+)-induced contraction at clamped levels of Ca2+. Phorbol 12,13-dibutyrate (PDBu, 10 microM) also increased the contractile force at a given level of Ca2+.(ABSTRACT TRUNCATED AT 400 WORDS)
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