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Biomedical subjects

H Yabu

Publications and source records attributed to H Yabu.

At least 37 records · Page 2Linked to original sources

Dual effect of phosphatase inhibitors on calcium channels in intestinal smooth muscle cells.

The effects of okadaic acid (OA) and calyculin A (CL-A), potent inhibitors of protein phosphatases type 1 (PP1) and type 2A (PP2A), on inward current carried by Ba2+ through voltage-dependent Ca2+ channel in guinea pig teniae coli smooth muscle cells were investigated using whole-cell patch-clamp technique. High concentrations of OA (5 x 10(-8)-5 x 10(-6) M) and CL-A (10(-9)-10(-7) M) dose dependently increased the inward current. The concentration producing apparent half-maximum enhancing effect values for OA and CL-A were 1.12 x 10(-7) and 1.78 x 10(-9) M, respectively. CL-A appeared to be approximately 100-fold more potent in increasing the inward current than OA. Lower concentrations of OA (10(-10)-2 x 10(-8) M) and CL-A (10(-11)-10(-9) M) decreased the inward current. The maximum inhibitory effects of OA and CL-A were observed at 10(-8) M OA and 5 x 10(-10) M CL-A, respectively. CL-A is approximately 100 times more effective inhibitor of PP1 than OA, and lower concentrations of OA and CL-A used in the present study inhibit PP2A activity, but they have no or little effect on PP1 activity (Ishihara, H., B. L. Martin, D. L. Brautigan, H. Karaki, H. Ozaki, Y. Kato, N. Fusetani, S. Watabe, K. Hashimoto, D. Uemura and D. J. Hartshorne. Biochem. Biophys. Res. Commun. 159: 871-877, 1989). In the absence of ATP in pipette solution, OA and CL-A did not affect the inward current.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

[Modulation of the voltage-dependent Ca2+ channels of smooth muscle by phosphorylation].

The voltage-dependent Ca2+ channels (VDC) in smooth muscle cell membranes are the major pathway by which Ca2+ enters the cell during contraction. It has been reported that VDC can be modulated by reversible channel protein phosphorylation and dephosphorylation reactions. In intestinal smooth muscle cell, muscarinic agents have been reported to increase the break-down of phosphatidylinositol 4,5-bisphosphate, which indicates that inositol-1,4,5-trisphosphate and diacylglycerol (DG) can be generated. DG activates protein kinase C. Carbachol (CCh), phorbol 12,13-dibutyrate and phosphatase inhibitors, okadaic acid and calyculin A increased the inward currents passing through the L-type VDCs. These effects were inhibited by protein kinase inhibitors, H-7 and staurosporine. The CCh effect was also inhibited by GDP beta S. Therefore, it seems possible that DG, a product of phosphatidylinositol break-down might mediate muscarinic effects on L-type VDC, the last via stimulation of protein kinase C, and that L-type VDC activity might be modulated by protein kinase C-mediated phosphorylation and protein phosphatase type-1-mediated dephosphorylation of the channel or related protein(s) in guinea pig taenia coli smooth muscle cells.

Animals↗

Contractile responses to electrical field stimulation and ATP in guinea-pig urinary bladder.

1. The responses of guinea-pig urinary bladder smooth muscle to intramuscular nerve stimulation were investigated by isometric tension recording. 2. The nerve mediated contractions, evoked by field stimulation with a stimulus of 0.5 msec duration, a frequency of 15 Hz and a voltage of 50 V, were inhibited by atropine (10(-6) M) and alpha, beta-MeATP (10(-4) M) to about 80% and 20%, respectively, of control. 3. Simultaneous application of atropine (10(-6) M) and alpha, beta-MeATP (10(-4) M) abolished the contraction evoked by selective nerve stimulation. 4. Terodiline (10(-4) M), one of Ca2+ antagonists, inhibited the contraction evoked by both nerve selective stimulation and exogenously applied ATP. 5. Based on these results we suggested that in guinea-pig urinary bladder, ATP was a predominant excitatory transmitter and the nerve-mediated contraction and exogenous ATP were mainly dependent upon Ca2+ influx.

Adenosine Triphosphate↗

Blocking action of terodiline on calcium channels in single smooth muscle cells of the guinea pig urinary bladder.

The blocking action of terodiline, a nonspecific organic Ca++ antagonist, on smooth muscle Ca++ channels of the guinea pig urinary bladder was investigated. Inward Ca++ currents were recorded from smooth muscle cells isolated from the urinary bladder using the whole-cell patch-clamp technique. In the absence of terodiline, a use-dependent reduction in the amplitude of inward Ca++ current was observed at a stimulus frequency of 0.2 Hz. When terodiline (1-10 microM) was applied, the use-dependent reduction was accelerated markedly, depending on the stimulus frequency. The blocking action of terodiline was also dose-dependent; the Kd value as measured at the end of 20 times repetitive stimulation at 0.2 Hz was 1.7 microM. In addition to such a use-dependent block, terodiline produced a hyperpolarizing shift in the steady-state inactivation curve. The results suggest that terodiline preferentially binds to the Ca++ channel in the open state and also in the inactivated state.

Action Potentials↗

Calyculin A increases voltage-dependent inward current in smooth muscle cells isolated from guinea pig taenia coli.

The effects of a potent phosphatase inhibitor, calyculin A (CL-A), on inward currents in guinea pig taenia coli smooth muscle cells were examined. CL-A increased the inward current, and this effect of CL-A was inhibited by a protein kinase C inhibitor, H-7, and by nifedipine. Phorbol 12,13-dibutyrate, an activator of protein kinase C, also increased the inward current and this effect was antagonized by H-7. These results suggest that in guinea pig taenia coli smooth muscle cells CL-A may facilitate the opening of the L-type Ca2+ channels through the protein kinase C-dependent phosphorylation system.

Animals↗

Two types of Ca channels in smooth muscle cells isolated from guinea-pig taenia coli.

1) Two types of voltage dependent Ca channels with different conductances and inactivation kinetics were identified from cell-attached patch clamp recordings. One type, with a larger conductance of 25pS, had a threshold of activation near -40mV and the mean current inactivated slowly. A second type of Ca channel, with a smaller conductance of 12pS channel, but the averaged mean current inactivated rapidly. 2) Cadmium ions inhibited the large conductance Ca channel currents, while the small conductance Ca channel currents was not blocked. 3) Large conductance Ca channel current selectively inhibited by dihydropyridine derivative, Nifedipine. Small conductance Ca channel current is abolished in dose dependent manner by pyrethroid insecticide, tetranethrine.

Animals↗

Multiple types of voltage-dependent Ca channels in mammalian intestinal smooth muscle cells.

(1) Whole-cell and single channel recording techniques have been applied to smooth muscle cells isolated from guinea-pig taenia coli to examine whether multiple types of Ca channels exist. (2) Whole-cell recordings under physiological Ca concentration (1.8 mM) revealed two current components with fast and slow inactivating kinetics. The fast inactivating component was present when cells were held at very negative potentials (-80 mV). It was insensitive to the dihydropyridine (DHP) derivative, nifedipine. In contrast, the slow inactivating component was present at less negative holding potentials. It was blocked by nifedipine. (3) The two current components were found to have closely similar voltage dependencies for activation. (4) These results suggest that the fast inactivating decay of the Ca current was mediated not only by the entry of Ca into the cell but also by a voltage-dependent process via a different type of Ca channel with fast inactivating kinetics. (5) Recordings from cell-attached membrane patches with 100 mM external Ba clearly showed the existence of multiple types of Ca channels with different conductances. (6) The large conductance channels (30 pS) activated at more positive potentials (0 mV) and their averaged current decayed much more slowly. The DHP Ca antagonist, nifedipine, inhibited the large conductance channels increasing the proportion of blank sweeps and reducing the averaged current. On the other hand, the DHP Ca-agonist, BayK 8644, increased the average current by increasing the mean open-times of the large conductance channels. The presence of micromolar Cd in the patch pipettes produced a flickering block of the large conductance channels.(ABSTRACT TRUNCATED AT 250 WORDS)

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗

Whole-cell and unitary Ca channel currents in mammalian intestinal smooth muscle cells: evidence for the existence of two types of Ca channels.

Whole-cell and single Ca channel currents were recorded in smooth muscle cells isolated from guinea-pig taenia coli to examine whether multiple types of Ca channels exist. Two different types of voltage-dependent Ca channels with different conductances and inactivation kinetics were identified from cell-attached patch clamp recordings using 50 mM Ba in the patch pipettes. One type of channel, with a large conductance of 25 pS, had a threshold of activation near -40 mV and the mean current reconstructed by averaging individual current responses inactivated slowly. A second type of channel, with a small conductance of 12 pS, had a similar threshold value to that of the 25 pS channel, but the averaged current inactivated rapidly. The steady state inactivation of the 12 pS channel was complete at a holding potential of -40 mV. We concluded that both channel types represent fast and slow inactivating voltage-dependent Ca channels which have been found in many excitable cells.

Animals↗

Calcium channel modulation by the dihydropyridine derivative Bay K 8644 in mammalian visceral smooth muscle cells.

1. Modulation of Ca channels by the dihydropyridine Ca agonist Bay K 8644 in guinea-pig taenia coli smooth muscle cells was investigated using the patch clamp technique. 2. Single Ca channel activity was obtained from cell-attached patch recordings with the use of pipettes filled with 50 mM Ba. Bath application of the drug markedly increased the opening probability of Ca channels. 3. The effect was found to be due to an increase in the mean opening times of Ca channels. Due to this increase, the mean current reconstructed by averaging individual current trace responses was markedly increased in the presence of Bay K 8644.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗

Relationship between length-tension relation and 20,000 dalton myosin light chain phosphorylation in guinea-pig taenia caeci.

1. Relationship between length-tension relation and phosphorylation of 20,000 dalton myosin light chain (LC20) in guinea-pig taenia caeci was investigated. 2. At in situ length (Lb), a good linear correlation was obtained between isometric tension and LC20 phosphorylation in high-K+-stimulated muscle. 3. In 100 mM K+-stimulated muscle, the active tension decreased at muscle lengths other than Lb, but no significant decrease in degree of LC20 phosphorylation was observed. 4. These results suggest that in guinea-pig taenia caeci, the major portion of the decrease in active tension at muscle lengths other than Lb is not due to a decrease in degree of activation.

Animals↗