Dual concentration-dependent effects of phorbol 12,13-dibutyrate on spontaneous and acetylcholine-induced electrical responses recorded from isolated circular smooth muscle of the guinea-pig stomach antrum

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  • Nakamura Eri
    Department of Physiology, Nagoya City University Medical School
  • Suzuki Hikaru
    Department of Physiology, Nagoya City University Medical School

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Intracellular recordings of electrical activity were made from circular smooth muscle cells in small segments of tissue isolated from the guinea-pig stomach antrum. Every cell that was impaled exhibited a rhythmic generation of slow potentials. Experiments were carried out to test the effects of three different concentrations (1, 10 and 100 nM) of phorbol 12, 13-dibutyrate (PDBu) on these slow potentials and on the responses produced by acetylcholine (ACh), in the presence of nifedipine and Nω-nitro-L-arginine (nitroarginine), known inhibitors of L-type Ca-channels and nitric oxide synthase, respectively. The resting membrane potential was -62 ± 7 mV, while the frequency and amplitude of the slow potentials were 1.6 ± 0.1 cycle per min (cpm) and 33 ± 1 mV, respectively. Application of 1 nM PDBu increased the frequency of slow potentials, with no significant change in the membrane potential and amplitude of slow potentials. At a concentration of 100 nM, PDBu depolarized the membrane by about 6 mV, and either decreased the amplitude and frequency of the slow potentials or abolished them. The amplitude and frequency of the slow potentials were not significantly changed in the presence of 10 nM PDBu. In the presence of chelerythrine (1-2 μM), a known inhibitor of protein kinase C (PKC), the increase in frequency of slow potentials by 1 nM PDBu and depolarization produced by 100 nM PDBu were not elicited. The increase in frequency of slow potentials by 100 nM ACh was inhibited by PDBu, in a concentration-dependent manner, and ACh-responses were abolished in the presence of 100 nM PDBu. These results indicate that PDBu has dual actions on the spontaneous activity of antral circular muscle, with low concentrations increasing and high concentrations inhibiting the frequency of the slow potentials. The former may be produced by activation of protein kinase C (PKC). As the ACh-induced excitation of slow potentials is inhibited by PDBu, a possible causal relationship between the inhibition and over-activation of PKC is considered.<br>

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