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Biomedical subjects

M Oike

Publications and source records attributed to M Oike.

49 records · Page 3Linked to original sources
PubMed1993

Electrogenic Na+/K(+)-transport in human endothelial cells.

Na+/K+ pump currents were measured in endothelial cells from human umbilical cord vein using the whole-cell or nystatin-perforated-patch-clamp technique combined with intracellular calcium concentration ([Ca2+]i) measurements with Fura-2/AM. Loading endothelial cells through the patch pipette with 40 mmol/l [Na+] did not induce significant changes of [Ca2+]i. Superfusing the cells with K(+)-free solutions also did not significantly affect [Ca2+]i. Reapplication of K+ after superfusion of the cells with K(+)-free solution induced an outward current at a holding potential of 0 mV. This current was nearly completely blocked by 100 mumol/l dihydroouabain (DHO) and was therefore identified as a Na+/K+ pump current. During block and reactivation of the Na+/K+ pump no changes in [Ca2+]i could be observed. Pump currents were blocked concentration dependently by DHO. The concentration for half-maximal inhibition was 21 mumol/l. This value is larger than that reported for other tissues and the block was practically irreversible. Insulin (10-1000 U/l) did not affect the pump currents. An increase of the intracellular Na+ concentration ([Na+]i) enhanced the amplitude of the pump current. Half-maximal activation of the pump current by [Na+]i occurred at about 60 mmol/l. The concentration for half-maximal activation by extracellular K+ was 2.4 +/- 1.2 mmol/l, and 0.4 +/- 0.1 and 8.7 +/- 0.7 mmol/l for Tl+ and NH4+ respectively. The voltage dependence of the DHO-sensitive current was obtained by applying linear voltage ramps. Its reversal potential was more negative than -150 mV. Pump currents measured with the conventional whole-cell technique were about four times smaller than pump currents recorded with the nystatin-perforated-patch method.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphate↗
PubMed1993

Development of preimplantation guinea-pig embryos in serum-free media.

This study was undertaken to determine how far preimplantation guinea-pig embryos can develop normally in vitro with serum-free media generally used for mouse (Whitten's medium, WM) and rabbit (Kane's medium, KM) embryos. Preimplantation guinea-pig embryos were recovered from naturally mated females at each day of pregnancy between Day 1 and Day 6, and were cultured in vitro with either WM or KM at 37 degrees C in 5% CO2 in air. One-cell (Day-1) and 2-cell (early Day 2) embryos cultured in WM developed to the 4-cell and morulae stages respectively, whereas the corresponding embryos cultured in KM arrested development at the 2-cell and 8-cell stages respectively. In contrast, 4-cell (late Day 2 and Day 3) embryos proceeded to the blastocyst stage in KM, but not in WM. Eight-cell embryos (Day 4) and morulae (Day 5) developed to blastocysts in either medium. However, hatching of the blastocysts and trophoblast outgrowth on the culture dish were supported only by KM even when collected at the blastocyst stage. The present study suggests that, in guinea-pig embryos, a distinct change in nutritional requirements occurs around the time of the second cell division and that, unlike in the mouse, trophoblast outgrowth in the guinea-pig requires no supplementation of serum or extracellular matrices.

Animals↗
PubMed1993

[The role of endothelin in adrenal medulla: identification of endothelin and its receptor in two cases of pheochromocytoma].

To clarify the role of a novel vasoactive peptide endothelin in endocrine hypertension, we examined endothelin (ET)-like immunoreactivity and the ET binding sites in the tissues of two pheochromocytoma. Case 1 was a 25-year-old woman who manifested multiple endocrine neoplasia (MEN) II b with left adrenal pheochromocytoma, and case 2 was a 48-year-old woman with familial bilateral pheochromocytoma. In both cases, fairly high amount of ET-like immunoreactivity was detected in the resected tumors. By competitive binding analysis, high affinity binding sites for 125I-ET-1 were also detected in the same resected tissues. Concomitant existence of ET and its receptor suggests that ET may play an important role in these tumors in modulating the secretion of catecholamines from the tumors by an autocrine/paracrine fashion.

Adrenal Gland Neoplasms↗
PubMed1992

Histamine H3-receptor activation augments voltage-dependent Ca2+ current via GTP hydrolysis in rabbit saphenous artery.

1. Actions of histamine on the voltage-dependent Ba2+(Ca2+) currents (IBa, ICa) were investigated using the whole-cell patch-clamp technique on dispersed smooth muscle cells from the rabbit saphenous artery. 2. Histamine (half-maximal dose, EC50 = 530 nM) augmented the IBa evoked by a brief depolarizing pulse (100 ms duration; to +10 mV from a holding potential of -80 mV) in a concentration-dependent manner. The maximum augmentation was obtained with 30 microM-histamine (1.29 times control). This augmentation of IBa was inhibited by the H3-antagonist, thioperamide (Ki = 30 nM, slope of the Schild plot = 1.0), but not by H1- or H2-antagonists (mepyramine or diphenhydramine, or cimetidine, respectively). 3. An H3-agonist, R alpha-methylhistamine (EC50 = 93 nM), also augmented IBa in a concentration-dependent manner at a holding potential of -80 mV and the maximum augmentation (1.25 times control) was obtained with 10 microM. This augmentation was also inhibited by thioperamide, but not by the above H1- and H2- antagonists. 4. Intracellularly applied 500 microM-guanosine 5'-triphosphate (GTP) enhanced, but 1 mM-guanosine 5'-O-(2-thiodiphosphate) (GDP beta S) abolished, the histamine-induced augmentation of IBa. When one of the non-hydrolysable GTP analogues, guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S; greater than 5 microM), guanylyl-imidodiphosphate (GMP-PNP; 200 microM) or guanylyl (beta, gamma-methylene)-diphosphonate (GMP-PCP; 1 mM) was intracellularly applied, the IBa amplitude evoked without the application of histamine was not affected, but the excitatory effect of histamine on IBa was reversed to an inhibition. Pre-treatment with pertussis toxin (PTX: 300 ng/ml and 3 micrograms/ml) did not modify the histamine-induced responses in the absence or presence of GTP gamma S. 5. 4 beta-Phorbol 12,13-dibutylate (PDBu) increased the amplitude of IBa. However, this action of PDBu was not enhanced by the application of GTP (500 microM) in the pipette, but additional application of histamine further increased the amplitude of IBa. Pre-treatment with a potent non-selective protein kinase inhibitor, 1-(5-isoquinolinesulphonyl)-2-methylpiperazine dihydrochloride (H-7; 100 microM), did not modify the histamine-induced current augmentation or inhibition observed in the presence or absence of intracellular GTP gamma S.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1990

Mechanisms of endothelin-induced augmentation of the electrical and mechanical activity in rat portal vein.

Actions of porcine endothelin (ET) on the electrical and mechanical activity of the rat portal vein were investigated by means of the intracellular microelectrode and isometric tension recording techniques, ET (greater than 0.1 nM) enhanced the amplitude and frequency of the spontaneous contractions which ceased in the presence of 100 nM dihydropyridine derivatives (nifedipine or nicardipine). ET (0.15 nM) increased the frequency of the spontaneous action potentials, with no change in the basal membrane potential. Higher concentrations of ET (greater than or equal to 0.3 nM) further depolarized the membrane potential and increased the spike frequency. After blocking the spontaneous action potentials with nifedipine (100 nM), ET still depolarized the membrane. The depolarization was associated with a reduction in the electrotonic potential and was blocked in a Na-deficient solution (15.5 mM) but not in Ca-free, K-deficient or Cl-deficient solutions. In a Na-deficient solution, ET still evoked action potentials without depolarization. In Ca-free solution, ET depolarized the membrane potential with small oscillations, which were blocked by nifedipine (100 nM). The results indicate that in the rat portal vein, ET enhances electrical and mechanical responses through activation of the dihydropyridine-sensitive and voltage-dependent Ca channels. Acceleration of the Ca entry induced by ET can occur with or without depolarization of the membrane and can enhance the pacemaking mechanism.

Animals↗
PubMed1990

Endothelin augments unitary calcium channel currents on the smooth muscle cell membrane of guinea-pig portal vein.

1. The effects of endothelin (ET) on the Ca2+ channel current in smooth muscle cells of the guinea-pig portal vein were investigated using the patch-clamp technique with whole-cell and cell-attached configurations. 2. ET augmented the macroscopic Ba2+ current in a dose-dependent manner; this effect was inhibited by nifedipine or Cd2+. Augmentation of the inward current by ET did not depend on the amplitude of the depolarizing pulse. Further, when the membrane potential was held at -60 mV, ET increased the amplitude of the Ba2+ inward current measured at the peak and end of the depolarizing pulse to the same extent. 3. By contrast, when the membrane potential was held at -80 mV, depolarizing pulses to potentials more negative than 0 mV produced greater augmentation of the inward current than did those more positive than 0 mV. Moreover, when a depolarizing pulse to below 0 mV was applied, ET increased the peak amplitude of the inward current more than the amplitude measured at the end of pulse. 4. Using the patch-clamp technique with cell-attached configuration, two types of unitary Ba2+ current with conductances of 22 and 12 pS were obtained in 50 mM-Ba2+ solution. Nifedipine inhibited both types of unitary channel current, but the sensitivity of the 22 pS Ca2+ channel to nifedipine was 20-fold higher than the 12 pS Ca2+ channel. 5. Bath application of ET prolonged the mean open time, reduced the number of sweeps in which no Ca2+ channel was opened ('blank' sweep), and increased the number of channel openings evoked by each depolarizing pulse without changes of conductance. As a consequence, ET increased the open probability of both channels. 6. Augmentation of the 12 pS channels by ET was seen only in the early phase of a depolarizing pulse (57 ms from the onset of 170 ms pulse), while augmentation of the 22 pS channels was seen during the entire period of a depolarizing pulse. 7. When ET was added to the pipette solution, the activity of both Ca2+ channels was increased. However, this effect was less frequently observed than when ET was applied in the bath. 8. These results suggest that ET augments both the nifedipine-sensitive and resistant Ca2+ channels in the smooth muscle cell membrane of the guinea-pig portal vein, but in different ways. Presumably, ET acts indirectly on the voltage-dependent Ca2+ channel.

Animals↗
PubMed1990

Dual action of FRC8653, a novel dihydropyridine derivative, on the Ba2+ current recorded from the rabbit basilar artery.

Actions of FRC8653 on the macroscopic and unitary Ba2+ currents were studied using the rabbit basilar artery. Application of (+/-)-FRC8653 (less than 1 microM) increased the amplitude of the inward current when depolarization pulses more negative than -10 mV were applied but inhibited it when depolarization was more positive than 0 mV (in each case from a holding potential of -80 mV). At a holding potential of -40 mV, (+/-)-FRC8653 (greater than 0.1 nM) consistently inhibited the inward current. (-)-FRC8653 (greater than 1 nM) inhibited the amplitude of the inward current evoked by a depolarizing pulse more positive than -10 mV (the holding potential being -80 mV). At the holding potential of -80 mV, but not at -40 mV, (+)-FRC8653 (1 microM) enhanced the current amplitude evoked by a depolarizing pulse more negative than -10 mV but inhibited the current evoked by a pulse more positive than 0 mV. (+/-)-FRC8653 shifted the voltage-dependent inhibition curves to the left, and the slope of the curve became steeper (test pulse of +10 mV). Two types of single Ca2+ channel currents (12 and 23 pS) were recorded from the basilar artery by the cell-attached patch-clamp method. Opening of the 12-pS channel occurred with a depolarizing pulse (-20 mV) from a holding potential of -80 mV, but not from one of -60 mV. (+)-FRC8653 activated, and (-)-FRC8653 inhibited, the 23-pS channel.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1990

Nicorandil opens a calcium-dependent potassium channel in smooth muscle cells of the rat portal vein.

The actions of nicorandil on ionic currents recorded from smooth muscle cells of the rat portal vein were investigated using patch-clamp technique. In the whole-cell voltage clamp experiments, nicorandil (greater than 30 microM) produced an outward current in a concentration-dependent manner. The reversal potential was -80 mV. The nicorandil-induced outward current was suppressed in the nominally Ca-free (2.5 mM Mn-containing) solution. Tetraethylammonium (greater than 1 mM) and 4-aminopyridine (greater than 1 mM) inhibited the nicorandil-induced outward current. Recordings of single K-channels showed that there were two types of K currents with different conductance (132 pS and 10 pS) in the smooth muscle cell membrane. Both channels were sensitive to intracellular Ca concentration. The 10-pS K-channel but not the 132-pS K-channel was activated by lowering the ATP concentration inside of the membrane. Low concentrations of tetraethylammonium (less than 1 mM) completely inhibited the 132-pS K-channel, but had no effect on the 10-pS K-channel. On the other hand, 4-aminopyridine (10 mM) inhibited the 10-pS K-channel but not 132-pS K-channel. Nicorandil (500 microM) applied to outside of the membrane increased the mean open-time and number of appearances but reduced slow component of the mean closed time of the 10-pS K-channel current, without affecting their amplitudes. Nicorandil (less than 500 microM) did not modify the 132-pS K-channel. From above results we concluded that nicorandil activates the Ca-dependent and ATP-sensitive K-channel in smooth muscle cells of the rat portal vein.(ABSTRACT TRUNCATED AT 250 WORDS)

4-Aminopyridine↗