Biomedical subjects

Y Ikeuchi

Publications and source records attributed to Y Ikeuchi.

At least 55 records · Page 3Linked to original sources

PubMed1995

ATP activates the potassium channel and enhances cytosolic Ca2+ release via a P2Y purinoceptor linked to pertussis toxin-insensitive G-protein in brain artery endothelial cells.

ATP produced whole-cell potassium currents in cultured endothelial cells of the bovine brain cortical arteries. P2 purinoceptor agonists evoked similar currents with the order of their potency: 2-methylthio ATP > ATP >> alpha, beta-methylene ATP > or = UTP > or = ADP >> AMP. ATP-evoked currents were inhibited by GDP beta S, but not by pertussis toxin (PTX). Furthermore, a phospholipase C (PLC) inhibitor, protein kinase C inhibitor, or cAMP-dependent protein kinase inhibitor had no effect on the currents. In addition to these effects, ATP enhanced intracellular free Ca2+ concentration ([Ca2+]i) in the presence and absence of extracellular Ca2+, and this [Ca2+]i increase was not inhibited by a PLC inhibitor. These results, thus, provide an indication that ATP activates the potassium channel and enhances [Ca2+]i via a P2Y purinoceptor linked to a PTX-insensitive G-protein, which is not involved in a PLC-mediated signaling pathway.

Adenine Nucleotides↗

PubMed1995

Dual effects of ATP on the potassium channel and intracellular Ca2+ release in smooth muscle cells of the bovine brain arteries.

ATP produced whole-cell potassium currents with a latency of 200 msec in cultured smooth muscle cells of bovine brain arteries. The currents were evoked via an ATP receptor linked to a pertussis toxin-insensitive G-protein, which is not involved in phospholipase C (PLC)-mediated signal transduction, and were not regulated by protein kinase A or C. In the cell-attached patches, ATP elicited single channel currents with two major classes of slope conductances and these currents were again induced within 100 msec after bath-application of ATP outside the patch pipette. In addition, ATP enhanced intracellular free Ca2+ concentration ([Ca2+]i) with no initiation time in the presence and absence of extracellular Ca2+, and this enhancement was not blocked by a PLC inhibitor. These results, thus, suggest that the activation of the potassium channel and the enhancement of [Ca2+]i induced by ATP may be caused by a direct action of the G-protein beta gamma subunits.

Adenosine Triphosphate↗

PubMed1995

Adenosine activates the potassium channel via a P2 purinoceptor but not via an adenosine receptor in cultured rat superior colliculus neurons.

The effect of adenosine on superior colliculus neurons was examined by whole-cell patch clamp recording. Adenosine elicited whole-cell potassium currents. A selective A1 or A2a adenosine receptor agonist induced no current and furthermore, adenosine-evoked currents were not inhibited by selective A1 or A2a adenosine receptor antagonists or a non-selective adenosine receptor (P1 purinoceptor) antagonist, indicating that the currents are not mediated by adenosine receptors. In contrast, P2 purinoceptor agonists, such as 2-methylthio ATP (2-MeSATP), ATP, ADP, and AMP, produced similar potassium currents, whereas alpha,beta-methylene ATP, beta,gamma-metylene ATP, or UTP had no response. The order of their potencies for the current amplitudes was 2-MeSATP > ADP > adenosine > ATP >> AMP and this order corresponds to that for the P2Y purinoceptor. These results, thus, suggest that adenosine exerts an inhibitory effect at the postsynaptic site by activating the potassium channel via a P2Y purinoceptor in superior colliculus neurons.

Adenosine↗

PubMed1995

A P2 purinoceptor activated by ADP in rat medullar neurons.

ADP evoked outwardly rectifying potassium currents with a latency of 0.6 s in cultured rat medullar neurons. Purinoceptor agonists, such as 2-methylthio ATP (2-MeSATP), ATP, AMP, alpha,beta-methylene ATP (alpha,beta-MeATP), and UTP, produced similar outward currents with the order of their potencies for current amplitudes: 2-MeSATP > ADP > ATP > or = alpha,beta-MeATP > or = AMP > UTP. This order corresponds to that for a subtype of P2Y purinoceptors. ADP-evoked currents were fully blocked by a broad G-protein inhibitor, guanosine-5'-O-(2-thiodiphosphate) (GDP beta S), whereas a G(i)/G(o)-protein inhibitor, pertussis toxin (PTX) had no effect. The currents were not affected by a phospholipase C (PLC) inhibitor, neomycin. Furthermore, a selective protein kinase C inhibitor, GF109203X or a selective cAMP-dependent protein kinase inhibitor, H-89 showed no effect on the currents. These results suggest that ADP activates the potassium channel via a P2Y purinoceptor linked to a PTX-insensitive G-protein and its channel regulation may be due to a direct action of the G-protein beta gamma subunits regardless of second messenger signaling cascades. Additionally, ADP enhanced intracellular free Ca2+ concentration ([Ca2+]i) both in the presence and absence of extracellular calcium, and this [Ca2+]i increase was not inhibited by neomycin. This provides an additional evidence that ADP binds to a subtype of P2Y purinoceptors, which is not involved in PLC stimulation.

Adenosine Diphosphate↗

PubMed1995

The P2Y purinoceptor-operated potassium channel is possibly regulated by the beta gamma subunits of a pertussis toxin-insensitive G-protein in cultured rat inferior colliculus neurons.

In whole-cell patches of inferior colliculus neurons, ADP evoked outwardly rectifying potassium currents with a latency of < 1 sec via P2Y purinoceptor. These currents were blocked by GDP beta S, while not by pertussis toxin (PTX). Additionally, a selective protein kinase C inhibitor, GF109203X, or a selective phospholipase A2 inhibitor, BPB, had no effect on the currents. In outside-out patches, ADP elicited single channel currents with same slope conductances as those obtained in cell-attached patches and the currents were again blocked by GDP beta S. The results presented here indicate that the P2Y purinoceptor-operated potassium channel in inferior colliculus neurons is activated only by a plasma membrane component, most likely by a direct coupling to the beta gamma subunits of a PTX-insensitive G-protein.

Adenosine Triphosphate↗

PubMed1995

Activation of endogenous protein kinase C enhances currents through alpha 1 and alpha 2 glycine receptor channels.

The effects of Ca2+ /phospholipid dependent (PKC) phosphorylation on the current amplitudes of alpha 1 and alpha 2 glycine receptors expressed in Xenopus oocytes were examined by whole cell voltage clamp recording. In studies using phorbol esters, PKC phosphorylation has been shown to reduce glycine-induced currents. Endogenous PKC activation by pretreatment with serum, however, enhanced the currents to around 140% in both alpha 1 and alpha 2 glycine receptors. This effect was completely blocked by a specific PKC inhibitor, GF109203X. Instead, treatment with a potent PKC activator, 12-O-tetradecanoylphorbol-13-acetate (TPA) revealed a decrease in glycine-gated channel currents. Thus, the present results demonstrate that glycine receptor phosphorylation mediated by endogenous pathway of PKC activation potentiates glycine-induced currents and phorbol esters may have a direct action on glycine receptor channels independent of PKC activation.

Animals↗

PubMed1995

Methylcobalamin induces a long-lasting enhancement of the postsynaptic field potential in hippocampal slices of the guinea pig.

Methylcobalamin (CH3-B12) enhanced postsynaptic field potential (PSP) elicited from hippocampal slices (around 180% at a maximum), lasting more than 1 h after washing out, while it had no effect on antidromic spike potential (AP). By contrast, cyanocobalamin containing CN- instead of CH3- showed no enhancement in the PSP amplitude, suggesting that CH3- plays an important role for the PSP enhancement by methylcobalamin. The treatment with N-methyl-D-aspartate (NMDA) receptor specific antagonist, DL-2-amino-5-phosphonovaleric acid (APV) inhibited methylcobalamin-induced PSP enhancement by 50%, suggesting that NMDA receptor is partially relevant to the formation of this enhancement. Glutamate release from electrically stimulated slices was not enhanced by the treatment with methylcobalamin, suggesting that methylcobalamin affects the postsynaptic sites in hippocampal neurons. In whole cell voltage clamp recordings using cultured hippocampal neurons, methylcobalamin increased the currents elicited by NMDA time-dependently. These results may indicate that CH3- is required to modulate the PSP amplitude at the postsynaptic site and at least, NMDA receptor is involved in the formation of long-lasting PSP potentiation induced by methylcobalamin.

2-Amino-5-phosphonovalerate↗

PubMed1995

ATP-evoked potassium currents in rat striatal neurons are mediated by a P2 purinergic receptor.

The effect of ATP on cultured striatal neurons was examined by whole cell voltage clamp recordings. ATP produced outwardly rectifying currents that reversed near the expected equilibrium potential for the potassium ion and the currents were blocked by intracellular Cs+. Purinergic receptor agonists such as ADP, AMP adenosine, and 2-methylthio ATP (2-MeSATP) also evoked similar outward currents. The order of their potencies was ATP >> 2-MeSATP > or = ADP > adenosine > AMP, corresponding to a P2 purinergic receptor. ATP-evoked currents were blocked by a specific protein kinase C (PKC) inhibitor, GF109203X. In addition, the intracellular perfusion of a G-protein inactivator, GDP beta S abolished ATP-induced currents, whereas pertussis toxin (PTX) had no effect on the currents. These results suggest that ATP activates a potassium channel in striatal neurons, which is regulated by protein kinase C (PKC) activation through a P2 purinergic receptor linked to PTX-insensitive G protein.

Adenosine Triphosphate↗

PubMed1995

Effects of high pressure treatment on Ca2+ release and Ca2+ uptake of sarcoplasmic reticulum.

To clarify the mechanism of pressure-induced meat tenderization or acceleration of meat conditioning, the pressure-induced morphological and biochemical changes in sarcoplasmic reticulum (SR), and Ca2+ release from SR in the rabbit skeletal muscle treated with high pressure (100-300 MPa, 5 min) were investigated in comparison with those of the SR from conditioned muscle. The destruction of the membrane structure of the SR expanded with increasing pressure applied to the muscle. Significant changes in the SDS-PAGE profile were not observed in the SR from the pressurized muscle up to 200 MPa, but a marked decrease of the ATPase protein and high-affinity Ca(2+)-binding protein were observed in the SR from the pressurized muscle at 300 MPa. The ATPase activities increased in the SR isolated from the muscle exposed to high pressure up to 200 MPa. When the muscle was pressurized at 300 MPa, the ATPase activity dropped to the same level with that of the SR from the untreated muscle. Ca2+ uptake ability of the SR vesicles measured using a fluorescent chelating reagent decreased with increasing pressure applied to the muscle. Ultrastructural studies showed that Ca2+, which was mainly localized in the SR region of the untreated fiber bundles, was translocated into myofibrillar space in the pressurized muscle. It is clear that a brief exposure of the muscle to high pressure causes considerable changes in membrane structure and biochemical function of SR as compared with those of SR in the muscle induced by conditioning.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphatases↗

PubMed1995

Cleavage of connectin by calpain and cathepsin D.

mu-Calpain quickly split the alpha-connectin in myofibrils into beta-connectin, and then produced a 1700-kDa component. Cathepsin D also split alpha-connectin into beta-connectin, further degrading it to fragments smaller than the 1700-kDa component with increasing incubation time. The action of cathepsin D on the connectin molecule was distinctly different from that of mu-calpain in terms of the splitting rate and manner. When freshly excised muscle was exposed to a temperature of 37 degrees C, complete disappearance of connectin (alpha, beta and 1700-kDa component) was observed within 36 h. In contrast, at 2 degrees C, about 75% of connectin was retained as beta-form even after 3 weeks. The present data suggest that the degradation of connectin in muscle might be caused by mu-calpain in the early stage of aging, and then with time, this action is replaced by m-calpain or cathepsin D. However, the possibility of other intrinsic proteases participating in the degradation of connectin still remains.

Animals↗

PubMed1994

Toluene inhibits synaptic transmission without causing gross morphological disturbances.

The effects of toluene on synaptic transmission and neuronal morphology were investigated using guinea pig hippocampal slices. Population spikes (PS) were elicited in granule cell layer by stimulation of the perforant path and antidromic potentials (AP) were evoked in the same locii by stimulation of mossy fibers. Toluene at a concentration of 1000 micrograms/ml completely depressed post-synaptic responses within 15 min but increased the AP to 140% of its original value. The PS recovered completely when washout was begun 10 and 20 min after onset of depression, but exhibited only partial recovery following a 40 min depression. There were no evident changes in staining of axons or cell bodies after toluene treatment. These results indicate that toluene at high concentrations (1000 micrograms/ml) inhibits synaptic transmission selectively and with longer exposures causes lasting physiological effects unaccompanied by gross morphological changes.

Animals↗

PubMed1994

The contribution of PKA to the excitatory mechanism of adenosine in guinea pig superior colliculus slices.

The involvement of cyclic AMP-dependent protein kinase (PKA) for excitatory action of adenosine on neurotransmission was investigated using superior colliculus slices. The postsynaptic potential was elicited in the superficial gray layer after optic layer stimulation. Application of dibutyryl cyclic AMP to the medium enhanced the postsynaptic potential, but additional application of adenosine at 100 microM did not change the amplitude. Treatment of slices with a non-selective protein kinase inhibitor, 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H-7) or a selective PKA inhibitor, N-[2-(methylamino)ethyl]-5-isoquinolinesulfonamide (H-8) blocked the excitatory action of adenosine. Forskolin enhanced the postsynaptic potential, which was counteracted by treatment with a highly selective PKA inhibitor, N-[2-(p-bromocinnamylamino)ethyl]-5-isoquinoline sulfonamide (H-89) and subsequent application of 100 microM adenosine did not potentiate the postsynaptic potential. These results indicate that the excitatory effect of adenosine on neurotransmission in the superior colliculus involves PKA system.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗

PubMed1994

Effects of calf breed on milk production and other economic traits of Holstein dams.

Production of purebred or crossbred feeder calves for beef, especially HolsteinxJapanese-Black (HxJB) and Japanese-Black (JB), from dairy cattle using artificial insemination or embryo transfer have been used widely in Japan. However, dairy farmers feel uneasy about the effects of calf breed on the economic traits of dams. In this study, those effects were investigated in 798 Holstein heifers bearing Holstein, HxJB, JB or other breed calves. The results of the least-squares ANOVA indicated the effects of calf breed to be significant for gestation length (P<0.01) and calf birth weight (P<0.01) but not for milk yield, fat yield, protein yield, peak yield, day of peak, number of artificial inseminations per pregnancy or days open. Frequency of dystocia was lower in dams bearing HxJB calves than in those bearing Holstein calves (P<0.05). There were no significant differences for frequencies of still birth, retained placenta and subsequent pregnancy. The present data suggest that the effects of calf breed do not place a serious problem on the economic traits of Holstein dams. In conclusion, it is indicated that the production of JB and HxJB feeder calves from Holstein dams does not result in a decrease in dam productivity.

Journal Article↗

PubMed1993

Excitatory and inhibitory effects of toluene on neural activity in guinea pig hippocampal slices.

To investigate the effect of toluene and its derivatives on neural activity, postsynaptic field potential (population spike, PS) of granule cells as well as antidromic potential (AP) and presynaptic fiber potential (FP) (perforant path) were recorded in the guinea pig hippocampal slices. Toluene at the concentration of 0.2 ng/ml to 20 micrograms/ml in the perfusion medium increased the amplitude of PS to 109-150%. Toluene also increased the amplitude of FP and AP, although the most remarkable enhancement was observed in the PS. However, toluene at the concentrations over 1000 micrograms/ml completely depressed the PS, whereas it increased the amplitude of AP to 130% of the original level. These results indicate that toluene has excitatory and inhibitory biphasic effects on neurotransmission in the hippocampal slices according to concentration applied.

Animals↗

PubMed1993

Effect of high hydrostatic pressure on the conversion of alpha-connectin to beta-connectin.

The factors affecting the conversion of alpha-connectin to beta-connectin induced by pressurization of muscle were investigated over a pressure range from 100 to 400 MPa by using SDS-PAGE and immunoblot analysis. When muscles were exposed to high pressures, the conversion of alpha-connectin to beta-connectin was the most pronounced at a pressure of 300 MPa, and the appearance of 1,200-kDa peptide accompanied by conversion of alpha- to beta-connectin was observed. Connectin was relatively resistant to degradation under a pressure of 400 MPa. The degradative products of beta-connectin reactive with mAb 2D4 were not observed. The effect of high pressure on connectin in isolated myofibrils was similar to that on connectin in muscle. Addition of leupeptin and E-64 to the isolated myofibrils resulted in the prevention of the degradation of connectin at each stage of the pressurization. The ability of calcium-activated protease (calpain) to hydrolyze connectin from alpha to beta gradually declined with increasing pressure. The results indicate that calpain is responsible for the pressure-induced conversion of alpha- to beta-connectin. The rate of this conversion is probably regulated by the pressure-dependent structural change of alpha-connectin and inactivation of calpain.

Animals↗

PubMed1993

[MRI of chronic toluene intoxication].

We examined MRI of 7 chronic "thinner" abusers sniffing mainly toluene for more than 5 years. MRI examination revealed cerebral atrophies in 5 out of 7 patients and hippocampal atrophies in 3 out of 7 patients. Five out of 7 patients showed low intensity signal in bilateral thalamus on T2 weighted images and 2 showed high intensity signal in internal capsule (especially posterior limb) on T2 weighted image together with the change of thalamus. The results of intelligence test for 6 patients showed significant impairment (56 +/- 17, mean +/- S.D.). SPECT with 99mTc-HMPAO of a patient showed non-selective diffuse low perfusion in cerebral cortex. Reduction of cerebral blood flow would be significant for understanding cerebral and hippocampal atrophies or the deterioration of the intelligence of chronic toluene abusers.

Adult↗

PubMed1992

Formation of oligosaccharides from lactitol by Aspergillus oryzae beta-D-galactosidase.

Six oligosaccharides were first formed from lactitol by a transgalactosylation reaction catalyzed by Aspergillus oryzae beta-D-galactosidase. From the results of methylation analysis, MS, and 1H- and 13C-NMR studies, it was concluded that these oligosaccharides are O-beta-D-galactopyranosyl-(1----4)-O-beta-D-galactopyranosyl-(1----4)-D- glucitol, O-beta-D-galactopyranosyl-(1----3)-O-beta-D-galactopyranosyl-(1----4)-D- glucitol, O-beta-D-galactopyranosyl-(1----4)-[O-beta-D-galactopyranosyl-(1----6)]- D- glucitol, O-beta-D-galactopyranosyl-(1----6)-O-beta-D-galactopyranosyl-(1---4)- glucitol, O-beta-D-galactopyranosyl-(1----4)-[O-beta-D-galactopyranosyl- (1----5)]-D-glucitol, and O-beta-D-galactopyranosyl-(1----4)-[O-beta-D-galactopyranosyl-(1----1)]- D-glucitol. The last three are newly observed oligosaccharides.

Aspergillus oryzae↗

PubMed1992

Instability of F-actin in the absence of ATP: a small amount of myosin destabilizes F-actin.

The effects of the neutral salt concentration, pH, and coexistence of myosin on the denaturation of F-actin without ATP at low temperature were studied using the DNase I inhibition assay. The percent denaturation of F-actin gradually increased with a decrease in pH from 8.0 to 5.2, on incubation for 2 weeks in the presence of 50 mM KCl at 0 degrees C. This change was much faster in 0.5 M KCl and more than 75% of the F-actin became denatured on incubation for 1 week at pH 5.2. The buffer composition was found to exert a strong influence on the denaturation of F-actin. That is, there was a tendency for the denaturation of F-actin at pH 6.0 to be faster in MES[2-(N-morpholino)ethanesulfonic acid]-NaOH buffer than in sodium phosphate buffer, the critical concentrations of actin in 0.5 M KCl being 0.31 mg/ml for MES-NaOH buffer and 0.15 mg/ml for sodium phosphate buffer. A sigmoidal relationship was found between the percent denaturation of F-actin and the KCl concentration added, the greatest change occurring at KCl concentrations between 0.25 and 0.75 M. The time courses of the denaturation of F-actin showed that the percent denaturation rose at first and that in time the rate of the increase decreased. In the case of pH 8.0 and 0.5 M KCl, it took about 1 week for the denaturation rate to begin to drop. The pH of 6.0 further promoted the instability of F-actin exposed to high KCl concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)

Actins↗