KINDAI UNIVERSITY


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WADA Tetsuyuki

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FacultyDepartment of Pharmacy / Graduate School of Medicine
PositionAssociate Professor
Degree
Commentator Guidehttps://www.kindai.ac.jp/meikan/809-wada-tetsuyuki.html
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Last Updated :2020/09/01

Education and Career

Education

  • Kindai University, Faculty of Pharmacy
  • Kindai University, Graduate School of Pharmacy
  • Kinki University, Graduate School, Division of Pharmaceutical Sciences

Research Activities

Research Areas

  • Life sciences, Pharmacology
  • Life sciences, Pharmaceuticals - health and biochemistry

Published Papers

  • Ethyl mercury induces Ca2+ uptake through the P2X7 receptor in a mouse cerebellar microglia cell line., Sakamoto Y, Nakamoto Y, Wada T Ichida, S Minami T, Toxicological and Environmental Chemistry., Toxicological and Environmental Chemistry., 96(6), 1 - 14, 2015 , Refereed
  • A search for the risk factors for hiccups and evaluation of antiemetic therapy in CDDP-based chemotherapy, using cluster analysis., Asano H, Watanabe M, Kawaguchi A, Yanae M, Funakami Y, Wada T, Matzno S, Yamazoe Y, Nishida S, Ichida S, Gan To Kagaku Ryoho., Gan To Kagaku Ryoho., 40(8), 1031 - 1036, Aug. 2013 , Refereed
  • Effects of the α1-adrenoceptor agonist phenylephrine on SART stress-induced orthostatic hypotension in rats, BioPsychoSocial Medicine, BioPsychoSocial Medicine, 4(13), Oct. 2010
  • Analytical study of interaction between spdider toxin and glutamate receptors., Peptide Science 2009, Peptide Science 2009, 349 - 352, 2010
  • Study directed toward the analysis of interaction between the spider toxin NPTX-594 and glutamate receptors., Peptide Science, Peptide Science, 2008, 401 - 404, 2009
  • Study directed toward the analysis of interaction between the spider toxin NPTX-594 and glutamate receptors., Peptide Science, Peptide Science, 2008, 401 - 404, 2008
  • Paclitaxel-2 '-ethylcarbonate prodrug can circumvent P-glycoprotein-mediated cellular efflux to increase drug cytotoxicity, Tadatoshi Tanino, Akihiro Nawa, Eisaku Kondo, Fumitaka Kikkawa, Tohru Daikoku, Tatsuya Tsurumi, ChenHong Luo, Yukihiro Nishiyama, Yuki Takayanagi, Katuhiko Nishimori, Seiji Ichida, Tetsuyuki Wada, Yasuyoshi Miki, Masahiro Iwaki, PHARMACEUTICAL RESEARCH, PHARMACEUTICAL RESEARCH, 24(3), 555 - 565, Mar. 2007
    Summary:Purpose: The aim of the study was to investigate whether 2'-ethylcarbonate-linked paclitaxel (TAX-2'-Et) circumvents P-glycoprotein (P-gp)-mediated cellular efflux and cytotoxicity enhanced by TAX-2'-Et activation within human culture cells transfected with a rabbit liver carboxylesterase (Ra-CES) cDNA. Materials and Methods: TAX-2'-Et transport was characterized in a human colon carcinoma cell line (Caco-2) and paclitaxel (TAX)-resistant ovarian carcinoma cells (SKOV3/TAX60). Expression of P-gp, multidrug resistance protein (MRP) 2 and Ra-CES was detected by Western blotting. Cytotoxicity against Ra-CES-expressing cells and cellular amount of TAX produced were determined by MTT assay and using HPLC, respectively. Results: Unlike rhodamine123 and TAX, TAX-2'-Et did not exhibit polarized transport in the Caco-2 cells in the absence or presence of verapamil. P-gp levels were expressed much higher in the SKOV3/TAX60 cells than in the Caco-2 cells. MRP2 protein was not detectable in the SKOV3/TAX60 cells. Uptake by the SKOV3/TAX60 cells was similar in quantity to the amount internalized by P-gp-negative SKOV3 cells. In the SKOV3/TAX60 cells, cellular uptake of TAX-2'-Et was not altered regardless of the absence or presence of verapamil. The cytotoxicity to the untransfected SKOV3 cells induced by TAX-2'-Et was significantly lower than that induced by TAX. In the Ra-CES-expressing SKOV3 line, the EC50 value of TAX (10.6 nM) was approximately four-fold higher than that of TAX-2'-Et (2.5 nM). Transfection of Ra-CES into another TAX-resistant ovarian carcinoma cells (KOC-7c) conferred a high level of TAX-2'-Et cytotoxicity via prodrug activation. The intracellular levels of TAX produced from TAX-2'-Et in the Ra-CES-positive KOC-7c cells significantly increased compared with the levels seen in exposure of the untransfected KOC-7c cells to TAX. Conclusions: TAX-2'-Et can circumvent P-gp-associated cellular efflux of TAX. TAX-2'-Et is converted into TAX by the Ra-CES, supporting its potential use as a theoretical GDEPT strategy for cancer cells expressing high levels of P-gp. The TAX-2'-Et prodrug efficiently increased the amount of intracellular TAX, which mediates tumor cell death.
  • Sodium influx through cerebral sodium-glucose transporter type 1 exacerbates the development of cerebral ischemic neuronal damage, Yui Yamazaki, Shinichi Harada, Tetsuyuki Wada, Teruki Hagiwara, Shigeru Yoshida, Shogo Tokuyama, EUROPEAN JOURNAL OF PHARMACOLOGY, EUROPEAN JOURNAL OF PHARMACOLOGY, 799, 103 - 110, Mar. 2017 , Refereed
    Summary:We recently reported that cerebral sodium-glucose transporter type 1 (SGLT-1) plays a role in exacerbation of cerebral ischemia. However, the mechanism by which cerebral SGLT-1 acts remains unclear. Here we demonstrated that sodium influx through cerebral SGLT-1 exacerbates cerebral ischemic neuronal damage. SGLT-specific sodium ion influx was induced using alpha-methyl-D-glucopyranoside (alpha-MG). Intracellular sodium concentrations in primary cortical neurons were estimated using sodium-binding benzofuran isophthalate fluorescence. SGLT-1 knockdown in primary cortical neurons and mice was achieved using SGLT-1 siRNA. The survival rates of primary cultured cortical neurons were assessed using biochemical assays 1 day after treatment. Middle cerebral artery occlusion (MCAO) was used to generate a focal cerebral ischemic model in SGLT-1 knockdown mice. The change in fasting blood glucose levels, infarction development, and behavioral abnormalities were assessed 1 day after MCAO. Treatment with 200 mM alpha-MG induced a continuous increase in the intracellular sodium concentration, and this increase was normalized after a-MG removal. Neuronal SGLT-1 knockdown had no effect on 100 mu M H2O2-induced neuronal cell death; however, the knockdown prevented the neuronal cell death induced by 17.5 mM glucose and the co-treatment of 100 mu M H2O2/8.75 mM glucose. Neuronal SGLT-1 knockdown also suppressed the cell death induced by alpha-MG alone and the co-treatment of 100 mu M H2O2/0.01 mM alpha-MG. Our in vivo results showed that the exacerbation of cerebral ischemic neuronal damage induced by the intracerebroventricular administration of 5.0 mu g alpha-MG/mouse was ameliorated in cerebral SGLT-1 knockdown mice. Thus, sodium influx through cerebral SGLT-1 may exacerbate cerebral ischemia-induced neuronal damage.
  • Sodium transport through the cerebral sodium-glucose transporter exacerbates neuron damage during cerebral ischaemia, Yui Yamazaki, Shinichi Harada, Tetsuyuki Wada, Shigeru Yoshida, Shogo Tokuyama, JOURNAL OF PHARMACY AND PHARMACOLOGY, JOURNAL OF PHARMACY AND PHARMACOLOGY, 68(7), 922 - 931, Jul. 2016 , Refereed
    Summary:ObjectivesWe recently demonstrated that the cerebral sodium-glucose transporter (SGLT) is involved in postischaemic hyperglycaemia-induced exacerbation of cerebral ischaemia. However, the associated SGLT-mediated mechanisms remain unclear. Thus, we examined the involvement of cerebral SGLT-induced excessive sodium ion influx in the development of cerebral ischaemic neuronal damage. Methods[Na+]i was estimated according to sodium-binding benzofuran isophthalate fluorescence. In the in vitro study, primary cortical neurons were prepared from fetuses of ddY mice. Primary cortical neurons were cultured for 5 days before each treatment with reagents, and these survival rates were assessed using biochemical assays. In in vivo study, a mouse model of focal ischaemia was generated using middle cerebral artery occlusion (MCAO). Key findingsIn these experiments, treatment with high concentrations of glucose induced increment in [Na+]i, and this phenomenon was suppressed by the SGLT-specific inhibitor phlorizin. SGLT-specific sodium ion influx was induced using a-methyl-D-glucopyranoside (a-MG) treatments, which led to significant concentration-dependent declines in neuronal survival rates and exacerbated hydrogen peroxide-induced neuronal cell death. Moreover, phlorizin ameliorated these effects. Finally, intracerebroventricular administration of a-MG exacerbated the development of neuronal damage induced by MCAO, and these effects were ameliorated by the administration of phlorizin. ConclusionsHence, excessive influx of sodium ions into neuronal cells through cerebral SGLT may exacerbate the development of cerebral ischaemic neuronal damage.
  • Ouabain exerts cytoprotection by diminishing the intracellular K+ concentration increase caused by distinct stimuli in human leukemic cells, Masayuki Takechi, Tetsuyuki Wada, Hideki Yagi, Takashi Masuko, Atsufumi Kawabata, JOURNAL OF PHARMACY AND PHARMACOLOGY, JOURNAL OF PHARMACY AND PHARMACOLOGY, 67(1), 126 - 132, Jan. 2015 , Refereed
    Summary:Objectives We tested if modulation of cytosolic K+ levels by ouabain, an inhibitor of Na+/K+-ATPase, exerts cytoprotection against distinct stressful stimuli in human leukemic cells. Methods The cytosolic K+, Na+ or Ca2+ levels and the cytotoxicity were evaluated by flow cytometry. Key findings Various cytotoxic chemicals and ultraviolet irradiation induced cell death and increased intracellular concentrations of K+, Na+ or Ca2+. Ouabain reduced the cytotoxicity and the elevation of cytosolic levels of K+ but not those of Na+ or Ca2+. Conclusions Our data thus suggest that elevated cytosolic K+ levels are associated with the cytotoxicity in response to distinct stressful stimuli and that ouabain exerts cytoprotection most probably by regulating intracellular K+ levels.
  • Differential Expression of the 5-HT3A and 5-HT3B Receptor in Differentiated NG108-15 Cells, Hajime Asano, Akinori Miyamoto, Mariko Nakao, Chiyuki Wakaki, Takuma Iida, Yoshinori Funakami, Tetsuyuki Wada, Seiji Ichida, NEUROCHEMICAL RESEARCH, NEUROCHEMICAL RESEARCH, 37(8), 1738 - 1746, Aug. 2012 , Refereed
    Summary:Previous work from this laboratory has shown that the serotonin (5-HT) induced response is significantly augmented in differentiated NG108-15 (NG) cells treated with dibutyryl cAMP (Bt(2)cAMP) due to qualitative and quantitative changes in the expression of the 5-HT3 receptor as demonstrated by specific [H-3] LY-278584 (a selective 5HT(3) receptor antagonist) binding. In this study, we investigated whether there is any change in the relative expression of the 5-HT3A and 5-HT3B subunits in NG cells differentiated following Bt(2)cAMP treatment cells. The major findings of this study were that the relative amount of 5-HT3B subunit mRNA in Bt(2)cAMP-treated NG cells 5 days following Bt(2)cAMP-treatment was greater than that in the untreated cells. In contrast, the relative expression of the 5-HT3B subunit protein in the Bt(2)cAMP-treated NG cells was much less than in the untreated cells, but the relative expression of the 5-HT3A subunit in the Bt(2)cAMP-treated NG cells was similar to the untreated cells. Therefore, no relationship between mRNA and protein expression for 5-HT3A and 5-HT3B subunits in Bt(2)cAMP treated and untreated NG cells were observed. It was also found that fluorescent intensity for the 5-HT3B subunit in the cell body of the Bt(2)cAMP treated and untreated NG cells gradually decreased from the day 1-5 after Bt(2)cAMP treatment. However, in specific areas such as the varicosity and nerve endings of the Bt(2)cAMP treated cells, staining intensity for the 5-HT3B subunits was stronger than in the untreated cells at the all time points, peaking at day 5 post-treatment. These results suggest that the augmented response induced by 5-HT acting via 5-HT3 receptors in differentiated NG cells may be due to changes in the relative amount of the 5-HT3B subunit, particularly the ratio and distribution of the 5-HT3A to (3B) subunits.
  • Specific Alternation of Rhythm in Temperature (SART) Stress-Induced Irritable Bowel Syndrome-Like Changes in Mice and Effects of Drugs, Yoshinori Funakami, Eiji Itoh, Taeko Hata, Tetsuyuki Wada, Seiji Ichida, BIOLOGICAL & PHARMACEUTICAL BULLETIN, BIOLOGICAL & PHARMACEUTICAL BULLETIN, 33(9), 1545 - 1549, Sep. 2010
    Summary:Stress is closely associated with the manifestation and progress of irritable bowel syndrome (IBS). For the purpose of establishing experimentally the relationship between IBS and stress, the transportation capacity of the small intestine in specific alternation of rhythm in temperature (SART)-stressed animals was studied using charcoal transportation method. The charcoal suspension was administered orally into the stomach of fasting mice. Mice were sacrificed after a certain time and %charcoal transit (%CT) of the small intestine was measured. The %CTs in SART-stressed mice were greater than those in unstressed or continuously cold-stressed mice. This increase in %CT remained for I week after discontinuation of SART stress loading. Cholinergic blockers decreased %CTs in SART-stressed mice. Increases in %CT by a cholinesterase inhibitor were less in SART-stressed mice than in unstressed mice. Increases of %CT in SART-stressed mice were suppressed by Neurotropine. These results suggested that the parasympathetic hypertonicity, not just cold, played a role in the increases in the transportation capacity in SART-stressed mice and that these animals can be a useful tool for elucidation of the mechanism of IBS.
  • Characteristics for Enhanced Response of Serotonin-Evoked Ion Dynamics in Differentiated NG108-15 Cells, Takashi Imanishi, Kayoko Matsushima, Akinori Kawaguchi, Hajime Asano, Yoshinori Funakami, Tetsuyuki Wada, Takashi Masuko, Shigeru Yoshida, Seiji Ichida, NEUROCHEMICAL RESEARCH, NEUROCHEMICAL RESEARCH, 34(5), 1011 - 1019, May 2009
    Summary:Characteristics for the up-regulated response in the concentration of intracellular calcium ion ([Ca(2+)] (i) ) and in the sodium ion (Na(+)) current by serotonin (5-HT) were investigated in differentiated neuroblastoma x glioma hybrid NG108-15 (NG) cells. The results for the changes in [Ca(2+)] (i) by 5-HT were as follows, (1) The 5-HT-induced Ca(2+) response was inhibited by 3 x 10(-9) M tropisetron (a 5-HT(3) receptor blocker), but not by other types of 5-HT receptor blockers; (2) The 5-HT-induced Ca(2+) response was mainly inhibited by calciseptine (a L-type Ca(2+) blocker), but not by other types of Ca(2+) channel blockers or 10(-7) M TTX (a voltage-sensitive Na(+) channel blocker); (3) When the extracellular Na(+) was removed by exchange with choline chloride or N-methyl-d-glucamine, the 5-HT-induced Ca(2+) response was extremely inhibited. The results for the 5-HT-induced Na(+) current by the whole cell patch-clamp technique were as follows, (1) The 5-HT-induced Na(+) current in differentiated cells was significantly larger than that in undifferentiated cells; (2) The ED(50) value for 5-HT-induced Na(+) current in undifferentiated and differentiated cells was almost the same, about 4 x 10(-6) M each other; (3) The 5-HT-induced Na(+) current was completely blocked by 3 x 10(-9) M tropisetron, but not by other 5-HT receptor antagonists and 10(-7) M TTX. These results suggested that 5-HT-induced Ca(2+) response in differentiated NG cells was mainly due to L-type voltage-gated Ca(2+) channels allowing extracellular Na(+) to enter via 5-HT(3) receptors, but not through voltage-gated Na(+) channels.
  • Hydrogen sulfide as a novel nociceptive messenger, Atsufumi Kawabata, Tsuyoshi Ishiki, Kelta Nagasawa, Shigeru Yoshida, Yumi Maeda, Tomoko Takahashi, Fumiko Sekiguchi, Tetsuyuki Wada, Seiji Ichida, Hiroyuki Nishikawa, PAIN, PAIN, 132(1-2), 74 - 81, Nov. 2007
    Summary:Hydrogen sulfide (H2S), an endogenous gasotransmitter, modulates various biological events such as inflammation in the mammalian body. The present study investigated possible involvement of H2S in peripheral nociceptive processing. Intraplantar (i.pl.) administration of NaHS, a H2S donor, produced prompt hyperalgesia in rats, accompanied by expression of Fos in the spinal dorsal horn. The H2S-evoked hyperalgesia was blocked by 5,5'-dithio-bis-(2-nitrobenzoic acid) (DTNB), an oxidizing agent, or ethosuximide and mibefradil, T-type Ca2+ channel inhibitors. L-Cysteine, an endogenous source for H2S, given i.pl., also elicited hyperalgesia, an effect being abolished by DL-propargylglycine (PPG) and beta-cyanoalanine (BCA), inhibitors of cystathionine-gamma-lyase, a H2S synthesizing enzyme. PPG and/or BCA partially inhibited the hyperalgesia induced by i.pl. lipopolysaccharide, an effect being reversed by i.pl. NaHS. In the patch-clamp study using undifferentiated NG108-15 cells that express T-type, but not other types, of Ca2+ channels, NaHS enhanced the currents through the T-type channels, an effect being blocked by DTNB. Thus, H2S appears to function as a novel nociceptive messenger through sensitization of T-type Ca2+ channels in the peripheral tissues, particularly during inflammation. (C) 2007 International Association for the Study of Pain. Published by Elsevier B.V. All rights reserved.
  • Enhancement of serotonin- and bradykinin-evoked calcium ion dynamics in differentiated NG108-15 cells, Takashi Imanishi, Kayoko Matsushima, Akinori Kawaguchi, Tetsuyuki Wada, Shigeru Yohida, Seiji Ichida, NEUROSCIENCE LETTERS, NEUROSCIENCE LETTERS, 405(1-2), 1 - 4, Sep. 2006
    Summary:Dynamic changes in the concentration of intracellular free-calcium ion ([Ca2+](i)) by carbachol (M) and neurotransmitter candidates was investigated in undifferentiated and differentiated neuroblastoma x glioma hybrid NG108-15 (NG)cells. [Ca2+], was increased in a dose-dependent manner by bradykinin (BK) and serotonin (5-HT) in differentiated NG cells, and the response to BK and 5-HT was significantly greater than that in undifferentiated NG cells. The EC50 value of BK was approximately 1.5 x 10(-8) M in both undifferentiated and differentiated NG cells. The EC50 value of 5-HT in differentiated NG cells was about 5 x 10(-6) M. The response to BK and 5-HT was almost completely inhibited by 10 nM Hoe140 (a BK B2 receptor antagonist) and 3 nM tropisetron (a 5-HT3 receptor antagonist), respectively. These results suggest that there are some mechanisms by which the response evoked by BK and 5-HT is up-regulated in differentiated NG cells. (c) 2005 Elsevier Ireland Ltd. All rights reserved.
  • Enhancement of veratridine-induced sodium dynamics in NG108-15 cells during differentiation, T Imanishi, K Matsushima, A Kawaguchi, T Wada, T Masuko, S Yoshida, S Ichida, BIOLOGICAL & PHARMACEUTICAL BULLETIN, BIOLOGICAL & PHARMACEUTICAL BULLETIN, 29(4), 701 - 704, Apr. 2006
    Summary:Developmental changes in dynamics of Na+ were studied in neuroblastoma x glioma hybrid NG108-15 cells during differentiation which was induced by dibutyryl cAMP (Bt(2)cAMP). Ratiometric Na+ imaging with a Na+-sensitive fluorescent dye SBFI (sodium-binding benzofuran isophthalate) revealed that the intracellular Na+ concentration ([Na+](i),) was not affected by the application of high K+ (60 mm) solution to either control or differentiated cells. When cells were exposed to 50,mu m veratridine (Vtd), an agonist of voltage-sensitive sodium channels (VSSCs), a significant increase in [Na+](i) was observed in differentiated but not in undifferentiated cells. Calculated mean [Na+], value increased from the basal 10.4 to 44.1 mm in response to 50 mu M Vtd. This Vtd response was reversibly inhibited by tetrodotoxin (TTX), a specific blocker for VSSCs, in a dose-dependent manner (IC50=1 nm). It is suggested that VSSCs in NG108-15 cells are sensitive to TTX and Vtd and that the number of VSSCs increases during differentiation.
  • Increased response to high KCl-induced elevation in the intracellular-Ca2+ concentration in differentiated NG108-15 cell and the inhibitory effect of the L-type Ca2+ channel blocker, calciseptine, T Imanishi, K Matsushima, A Kawaguchi, T Wada, S Yoshida, S Ichida, NEUROCHEMICAL RESEARCH, NEUROCHEMICAL RESEARCH, 31(1), 33 - 40, Jan. 2006
    Summary:Characteristics of the increasing effect for the concentration of intracellular calcium ions ([Ca2+](i)) by high-KCl application were investigated in the neuroblastoma x glioma hybrid NG108-15 cell line ( NG108-15 cells). The present study confirmed that the increasing effect of [ Ca2+](i) by high-KCl application in single NG108-15 cells, differentiated with dibutyryl cAMP ( Bt(2)cAMP), was significantly enhanced, compared to undifferentiated cells. The following observations were made at first: ( 1) The response to high-KCl application, in both undifferentiated and differentiated cells, was significantly inhibited by calciseptine ( CaS), an L-type Ca2+ channel blocker, but not by N-, P- and R- type Ca2+ channel blockers. The IC50 values for CaS in both undifferentiated and differentiated cell was almost identical. ( 2) The inhibitory effect of CaS was irreversible. ( 3) The increasing effect for [ Ca2+](i) by high-KCl application was completely dependent on the presence of extracellular calcium ions. ( 4) The increased [ Ca2+](i) by high-KCl application under a plateau concentration was quickly decreased to basal levels when the high- KCl solution was exchanged for a high- KCl solution containing EGTA ( without CaCl2). Together, these results suggest that the enhancement of the response effect of [ Ca2+](i) by high-KCl application in differentiated single NG108-15 cells was mainly due to the quantitative increase of L-type voltage-sensitive calcium channels ( VSCCs), which were irreversibly inhibited by CaS.
  • Signal transduction for proteinase-activated receptor-2-triggered prostaglandin E-2 formation in human lung epithelial cells, N Kawao, M Nagataki, K Nagasawa, S Kubo, K Cushing, T Wada, F Sekiguchi, S Ichida, MD Hollenberg, WK MacNaughton, H Nishikawa, A Kawabata, JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS, JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS, 315(2), 576 - 589, Nov. 2005
    Summary:We investigated proteinase-activated receptor-2 (PAR(2))-triggered signal transduction pathways causing increased prostaglandin E-2 (PGE(2)) formation in human lung-derived A549 epithelial cells. The PAR(2) agonist, SLIGRL-NH2 (Ser-Leu-Ile-Gly-Arg-Leu-amide), evoked immediate cytosolic Ca2+ mobilization and delayed (0.5-3 h) PGE(2) formation. The PAR(2)-triggered PGE(2) formation was attenuated by inhibition of the following signal pathway enzymes: cyclooxygenases 1 and 2 (COX-1 and COX-2, respectively), cytosolic Ca2+-dependent phospholipase A(2) (cPLA(2)), the mitogenactivated protein kinases (MAPKs), mitogen-activated protein kinase/extracellular signal-regulated kinase kinase (MEK)-extracellular signal-regulated kinase (ERK) and p38 MAPK, Src family tyrosine kinase, epidermal growth factor (EGF) receptor tyrosine kinase (EGFRK), and protein kinase C (PKC), but not by inhibition of matrix metalloproteinases. SLIGRL-NH2 caused prompt ( 5 min) and transient ERK phosphorylation, blocked in part by inhibitors of PKC and tyrosine kinases but not by an EGFRK inhibitor. SLIGRL-NH2 2 also evoked a relatively delayed (15 min) and persistent (30 min) phosphorylation of p38 MAPK, blocked by inhibitors of Src and EGFRK but not by inhibitors of COX-1 or COX-2. SLIGRL-NH2 elicited a Src inhibitor-blocked prompt (5 min) and transient phosphorylation of the EGFRK. SLIGRL-NH2 up-regulated COX-2 protein and/or mRNA levels that were blocked by inhibition of p38 MAPK, EGFRK, Src, and COX-2 but not MEK-ERK. SLIGRL-NH2 also caused COX-1-dependent up-regulation of microsomal PGE synthase-1 (mPGES-1). We conclude that PAR(2)-triggered PGE(2) formation in A549 cells involves a coordinated up-regulation of COX-2 and mPGES-1 involving cPLA(2), increased cytosolic Ca2+, PKC, Src, MEK-ERK, p38 MAPK, Src-mediated EGF receptor trans-activation, and also metabolic products of both COX-1 and COX-2.
  • Effects of calmodulin and Ca2+ channel blockers on omega-conotoxin GVI a binding to crude membranes from alpha(1B) subunit (Ca(v)2.2) expressed BHK cells and mice brain lacking the alpha(1B) subunits, T Wada, T Imanishi, A Kawaguchi, MX Mori, Y Mori, K Imoto, S Ichida, NEUROCHEMICAL RESEARCH, NEUROCHEMICAL RESEARCH, 30(8), 1045 - 1054, Aug. 2005
    Summary:Characteristics for the specific binding of I-125-omega-CTX GVIA and I-125-omega-CTX MVIIC to crude membranes from BHKN101 cells expressing the alpha(1B) subunits of Ca(v)2.2 channels and from mice brain lacking the alpha(1B) subunits of Ca(v)2.2 channels, particularly, the effects of CaM and various Ca2+ channel blockers on these specific bindings were investigated. Specific binding of I-125-omega-CTX GVIA to the crude membranes from BHKN101 cells was observed, but not from control BHK6 cells. omega-CTX GVIA, omega-CTX MVIIC and omega-CTX SVIB inhibited the specific binding of I-125-omega-CTX GVIA to crude membranes from BHKN101 cells, and the IC50 values for omega-CTXGVIA, omega-CTX MVIIC and omega-CTX SVIB were 0.07, 8.5 and 1.7 nM, respectively. However, omega-agatoxin IVA and calciseptine at concentrations of 10(-9)-10(-6) M did not inhibit specific binding. Specific binding was also about 80% inhibited by 20 mu g protein/ml CaM. The amount of I-125-omega-CTX GVIA (30 pM) specifically bound to membranes from brain of knockout mice lacking alpha(1B) subunits of Ca(v)2.2 channels was about 30% of that to the crude membranes from brain of wild-type. On the other hand, specific binding of I-125-omega-CTX MVIIC (200 pM) was observed on the crude membranes of both BHKN101 and control BHK6 cells. The specific binding of I-125-omega-CTX MVIIC (200 pM) was not inhibited by omega-CTX GVIA and omega-CTX SVIB, and also omega-Aga IVA and calciseptine at concentrations of 10(-9)-10(-7) M, although specific binding was almost completely dose dependently inhibited by non-radiolabeled omega-CTX MVIIC (IC50 value was about 0.1 nM). 20 mu g protein/ml CaM did not inhibit specific binding. Therefore, these results suggest that BHKN101 cells have a typical Ca(v)2.2 channels which are also inhibited by CaM and have not specific binding sites for omega-CTX MVIIC, although omega-CTX MVIIC is a blocker for both Ca(v)2.1 (alpha(1A;) P/Q-type) and Ca(v)2.2 channels.

Conference Activities & Talks

  • Utility of the human organs observation as part of the humanistic education,   2017 03
  • RELATIONSHIP BETWEEN INTRACELLULAR K+ CONCENTRATION INCREASE AND CELL DIFFERENTIATION, T. Wada, M. Takechi, K. Sakai, K. Ueshiba, Y. Funakami, S. Yoshida, S. Ichida, THE FEDERATION OF EUROPEAN NEUROSCIENCE SOCIETIES (FENS) 9th FENS,   2014 07
  • A CHARACTERISTIC OF THE FUNCTION OF THE HYPOTHALAMUS-PITUITARY-ADRENAL AXIS IN AGED AND CHRONIC STRESSED MICE, Y. Funakami, T. Miyamoto, T. Iida, K. Sakai, K. Ueshiba, T. Wada, THE FEDERATION OF EUROPEAN NEUROSCIENCE SOCIETIES (FENS) 9th FENS,   2014 07
  • CHRONIC STRESS DECREASES THE FUNCTION OF THE HYPOTHALAMUS-PITUITARY-ADRENAL AXIS VIA GLUCOCORTICOID RECEPTORS IN MICE, S. Ichida, Y. Funakami, T. Miyamoto, T. Iida, K. Sakai, K. Ueshiba, T. Wada, THE FEDERATION OF EUROPEAN NEUROSCIENCE SOCIETIES (FENS) 9th FENS,   2014 07
  • Chronic stress attenuates neuronal activity of the brain stress circuit and the function of the hypothalamus-pituitary-adrenal axis in mice., Y Funakami, T Miyamoto, T Iida, K Kurokawa, K Sakai, K Ueshiba, M Nagano, T Wada, Y Shigeyoshi, S Ichida, Pharmacology 2013 London,   2013 12
  • Diazepam improves the functional disorder of the alveolar macrophage caused by chronic stress in mice., S Ichida, Y Funakami, C Wakaki, K Ueshiba, M Nakao, K Sakai, T Wada, Pharmacology 2013 London,   2013 12
  • Changes in intracellular concentration of potassium in the neuronal differentiation,   2013 09
  • Effects of Diazepam on c-Fos expression in hypothalamus of SART-stressed mice, Y Funakami, T Iida, T Miyamoto, M Kishimoto, K Toyoda, Y Taniguchi, S Ohura, Y Okabe, H, Asano,T Wada, M Nagano, Y Shigeyoshi, S Ichida, Neuro2012 Nagoya,   2012 09
  • Characteristics for response to serotonin and serotonin 3A/B receptor localization at nerve endings, in NG108-15 cells at the early phase after Bt2cAMP treatment,   2011 03
  • Change for 3A and 3B subunits of 5-HT3 receptors in differentiated NG108-15 cells, The 38th annual meeting of the Society for Neuroscience,   2008 11 , The 38th annual meeting of the Society for Neuroscience
  • Change for 3A and 3B subunits of 5-HT3 receptors during neural differentiation, Seiji Ichida, Hajime Asano, Yoshinori Funakami, Tetsuyuki Wada, Shigeru Yoshida,   2008 07
  • Zinc induces calcium uptake in rat pancreatic AR42J cells., T. Minami, T. Wada, H. Sugito, A. Kubo, S. Ichida, S. Yoshida, International Congress of Toxicology,Canada POSTER,   2007 06
  • Characteristic of serotonin-induced Na+ current in undifferentiated and differentiated NG108-15 cells, Neuroscience2006,   2006 10 , Neuroscience2006
  • Hydrogen sulfide sensitizes T-type calcium channels via redox modulation., Neuroscience 2006,   2006 10 , Neuroscience 2006
  • Decreased Intestinal Detoxification Functions in an Absorption Process in Adjuvant-Induced Arthritis Rats,   2005
  • Characteristics of N-type calcium channels stably expressed in BHK N101cells.,   2004 10
  • Characteristics of calcium influx elicited by high-KCl in differentiated and undifferentiated NG108-15 cells, Neuroscience2004,   2004 10 , Neuroscience2004
  • Signal transduction mechanisms for prostanoid formation caused by protease-activated receptor-2 activation in human lung epithelial cells., XVIIth International Congress on Fibrinolysis and Proteolysis,   2004 03 , XVIIth International Congress on Fibrinolysis and Proteolysis
  • Study for the characteristic of specific omega-conotoxin MVIIC binding on CaV2.1 and CaV2.2 channels fixed with antibodies and on membranes from N101 cells induced by CaV2.2 channel ?1 subunits,   2003 10

Misc

  • Inhibitory effects of tetrahydroisoquinoline derivatives on Ca2+ and Na+ channels in crude nerve endings, Y Zhang, J Abe, T Wada, S Ichida, GY Xu, BIOLOGICAL & PHARMACEUTICAL BULLETIN, 23, 3, 375, 378,   2000 03 , 10.1248/bpb.23.375
    Summary:Semi-synthetic tetrahydroisoquinoline derivatives prepared from natural alkaloids, possess Ca2+ antagonistic properties. These derivatives significantly blocked KCl-stimulated Ca2+ uptake (In chick and rat crude nerve endings) which fan be partially inhibited by the selective N-type Ca2+ channel blocker omega-conotoxin GVIA or the selective P-type Ca2+ channel blocker omega-agatoxin IVA. Moreover, PX42 (10 mu M; for the tetrahydroisoquinoline compounds in this study) could inhibit the activity of calmodulin-dependent phosphodiesterase and block veratridine-induced (or tetrodotoxin-sensitive) Na+ uptake. The possible mechanism(s) of non-selective inhibition of ion channels of PX42 is discussed.
  • Calcium/calmodulin inhibits the binding of specific [I-125]omega-conotoxin GVIA to chick brain membranes, S Ichida, J Abe, YA Zhang, K Sugihara, K Imoto, T Wada, H Sohma, NEUROCHEMICAL RESEARCH, 25, 3, 335, 340,   2000 03 , 10.1023/A:1007532818918
    Summary:The effect of Ca2+/calmodulin (CaM) on the specific binding of [I-125]omega-conotoxin GVIA (I-125-omega-CTX) to crude membranes from chick brain was investigated. When we examined the effects of the activation of various endogenous protein kinases on specific [I-125]omega-CTX binding to crude membranes, we observed that Ca2+/CaM had an inhibitory effect regardless of whether or not the standard medium contained ATP (0.5 mM). Ca2+/CaM also had an inhibitory effect in a simple binding-assay medium containing HEPES-HCl buffer, BSA, Ca2+ and CaM,and this effect was dependent on the concentration of Ca2+. The effect of Ca2+/CaM was attenuated by the CaM antagonists W-7 and CaM-kinase II fragment (290-309). An experiment with modified ELISA using purified anti omega-CTX antibody indicated that Ca2+/CaM did not affect the direct binding of [I-125]omega-CTX and CaM. These results suggest that Ca2+/CaM either directly or indirectly affects specific [I-125]omega-CTX binding sites, probably N-type Ca2+ channels in crude membranes from chick whole brain.
  • Proportions of Ca2+ channel subtypes in chick or rat P2 fraction and NG108-15 cells using various Ca2+ blockers, Zhang Yu-an, Takashi Imanishi, Tetsuyuki Wada, Seiji Ichida, Neurochemical Research, 24, 8, 1059, 1066,   1999 , 10.1023/A:1021065028647
    Summary:The proportions of calcium (Ca2+) channel subtypes in chick or rat P2 fraction and NG 108-15 cells were investigated using selective L-, N-, P- and P/Q- type Ca2+ channel blockers. KCl-stimulated 45Ca2+ uptake by chick P2 fraction was blocked by 40~50% using N-type Ca2+ channel blockers [ω- conotoxin GVIA, aminoglycoside antibiotics and dynorphin A(1-13)], but was not inhibited by P- or P/Q-type blockers (ω-agatoxin IVA or ω-conotoxin MVIIC). On the other hand, KCl-stimulated 45Ca2+ uptake by rat P2 fraction was blocked by 30~40% using P- or P/Q-type Ca2+ channel blockers, but was not inhibited by N-type Ca2+ channel blockers. The L-type Ca2+ channel blockers 1,4-dihydropyridines, diltiazem and verapamil, but not calciseptine (CaS), inhibited both KCl-stimulated 45Ca2+ uptake and veratridine-induced 22Na+ uptake by chick or rat P2 fraction with similar IC50 values. CaS did not have any effect on 45Ca2+ uptake by either chick or rat P2 fraction. In NG 108-15 cells, CaS, ω-agatoxin IVA and ω- conotoxin MVIIC, but not ω-conotoxin GVIA, inhibited KCl-stimulated 45Ca2+ uptake by 30-40%. Various combinations of these Ca2+ channel blockers had no significant additional effects in chick or rat P2 fraction or NG 108-15 cells. These findings suggest that KCl-stimulated 45Ca2+ uptake by chick or rat P2 fraction and NG 108-15 cells is a convenient and useful model for screening whether or not natural or synthetic substances have selective effects as L-, N-, P-, or P/Q- type Ca2+ channel antagonists or agonists.

Research Grants & Projects

  • Study on characteristics of Specific 125I-ω-conotoxin GVTA binding sites