16 results on '"Mika, Horiguchi"'
Search Results
2. A simple and rapid method for standard preparation of gas phase extract of cigarette smoke.
- Author
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Tsunehito Higashi, Yosuke Mai, Yoichi Noya, Takahiro Horinouchi, Koji Terada, Akimasa Hoshi, Prabha Nepal, Takuya Harada, Mika Horiguchi, Chizuru Hatate, Yuji Kuge, and Soichi Miwa
- Subjects
Medicine ,Science - Abstract
Cigarette smoke consists of tar and gas phase: the latter is toxicologically important because it can pass through lung alveolar epithelium to enter the circulation. Here we attempt to establish a standard method for preparation of gas phase extract of cigarette smoke (CSE). CSE was prepared by continuously sucking cigarette smoke through a Cambridge filter to remove tar, followed by bubbling it into phosphate-buffered saline (PBS). An increase in dry weight of the filter was defined as tar weight. Characteristically, concentrations of CSEs were represented as virtual tar concentrations, assuming that tar on the filter was dissolved in PBS. CSEs prepared from smaller numbers of cigarettes (original tar concentrations ≤ 15 mg/ml) showed similar concentration-response curves for cytotoxicity versus virtual tar concentrations, but with CSEs from larger numbers (tar ≥ 20 mg/ml), the curves were shifted rightward. Accordingly, the cytotoxic activity was detected in PBS of the second reservoir downstream of the first one with larger numbers of cigarettes. CSEs prepared from various cigarette brands showed comparable concentration-response curves for cytotoxicity. Two types of CSEs prepared by continuous and puff smoking protocols were similar regarding concentration-response curves for cytotoxicity, pharmacology of their cytotoxicity, and concentrations of cytotoxic compounds. These data show that concentrations of CSEs expressed by virtual tar concentrations can be a reference value to normalize their cytotoxicity, irrespective of numbers of combusted cigarettes, cigarette brands and smoking protocols, if original tar concentrations are ≤15 mg/ml.
- Published
- 2014
- Full Text
- View/download PDF
3. A Sialylated Voltage-Dependent Ca
- Author
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Yoichiro, Fujioka, Shinya, Nishide, Toyoyuki, Ose, Tadaki, Suzuki, Izumi, Kato, Hideo, Fukuhara, Mari, Fujioka, Kosui, Horiuchi, Aya O, Satoh, Prabha, Nepal, Sayaka, Kashiwagi, Jing, Wang, Mika, Horiguchi, Yuko, Sato, Sarad, Paudel, Asuka, Nanbo, Tadaaki, Miyazaki, Hideki, Hasegawa, Katsumi, Maenaka, and Yusuke, Ohba
- Subjects
Mice, Inbred BALB C ,Calcium Channels, L-Type ,Hemagglutinin Glycoproteins, Influenza Virus ,Madin Darby Canine Kidney Cells ,Mice, Inbred C57BL ,Mice ,Dogs ,HEK293 Cells ,Orthomyxoviridae Infections ,A549 Cells ,Influenza A virus ,COS Cells ,Chlorocebus aethiops ,Influenza, Human ,Animals ,Humans ,HeLa Cells - Abstract
Influenza A virus (IAV) infection is initiated by the attachment of the viral glycoprotein hemagglutinin (HA) to sialic acid on the host cell surface. However, the sialic acid-containing receptor crucial for IAV infection has remained unidentified. Here, we show that HA binds to the voltage-dependent Ca
- Published
- 2017
4. Agonist-promoted Ubiquitination Differentially Regulates Receptor Trafficking of Endothelin Type A and Type B Receptors
- Author
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Yoichiro Fujioka, Takahiro Horinouchi, Yusuke Ohba, Sarita Karki, Tsunehito Higashi, Koji Terada, Yosuke Mai, Prabha Nepal, Mika Horiguchi, Akimasa Hoshi, Chizuru Hatate, Takuya Harada, and Soichi Miwa
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media_common.quotation_subject ,education ,Blotting, Western ,Mutant ,Biology ,Biochemistry ,Endothelin ,Ubiquitin ,Lysosome ,medicine ,Humans ,Phosphorylation ,Extracellular Signal-Regulated MAP Kinases ,Internalization ,Receptor ,Molecular Biology ,health care economics and organizations ,Late endosome ,media_common ,Trafficking ,Microscopy, Confocal ,Endothelin-1 ,Cell Membrane ,Ubiquitination ,rab7 GTP-Binding Proteins ,Cell Biology ,Receptor, Endothelin A ,Receptor, Endothelin B ,Endocytosis ,Cell biology ,Protein Transport ,HEK293 Cells ,medicine.anatomical_structure ,rab GTP-Binding Proteins ,Mutation ,cardiovascular system ,biology.protein ,Lysosomes ,Intracellular - Abstract
Two types of G protein-coupled receptors for endothelin-1 (ET-1), ET type A receptor (ETAR) and ETBR, closely resemble each other, but upon ET-1 stimulation, they follow totally different intracellular trafficking pathways; ETAR is recycled back to plasma membrane, whereas ETBR is targeted to lysosome for degradation. However, the mechanisms for such different fates are unknown. Here we demonstrated that ETBR but not ETAR was ubiquitinated on the cell surface following ET-1 stimulation and that ETBR was internalized and degraded in lysosome more rapidly than ETAR. The mutant ETBR (designated "5KR mutant") in which 5 lysine residues in the C-tail were substituted to arginine was not ubiquitinated, and its rates of internalization and degradation after ET-1 stimulation became slower, being comparable with those of ETAR. Confocal microscopic study showed that following ET-1 stimulation, ETAR and 5KR mutant of ETBR were co-localized mainly with Rab11, a marker of recycling endosome, whereas ETBR was co-localized with Rab7, a marker of late endosome/lysosome. In the 5KR mutant, ET-1-induced ERK phosphorylation and an increase in the intracellular Ca(2+) concentration upon repetitive ET-1 stimulation were larger. A series of ETBR mutants (designated "4KR mutant"), in which either one of 5 arginine residues of the 5KR mutant was reverted to lysine, were normally ubiquitinated, internalized, and degraded, with ERK phosphorylation being normalized. These results demonstrate that agonist-induced ubiquitination at either lysine residue in the C-tail of ETBR but not ETAR switches intracellular trafficking from recycling to plasma membrane to targeting to lysosome, causing decreases in the cell surface level of ETBR and intracellular signaling.
- Published
- 2014
5. Endothelin-1 activates extracellular signal-regulated kinases 1/2 via transactivation of platelet-derived growth factor receptor in rat L6 myoblasts
- Author
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Takahiro Horinouchi, Soichi Miwa, Akimasa Hoshi, Yosuke Mai, Tsunehito Higashi, Takuya Harada, Chizuru Hatate, Tsunaki Higa, Mika Horiguchi, Koji Terada, and Prabha Nepal
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Dynamins ,Transcriptional Activation ,MAPK/ERK pathway ,Gene Expression Regulation, Enzymologic ,General Biochemistry, Genetics and Molecular Biology ,Cell Line ,Myoblasts ,Transactivation ,Growth factor receptor ,Animals ,Insulin ,Receptors, Platelet-Derived Growth Factor ,Phosphorylation ,General Pharmacology, Toxicology and Pharmaceutics ,Extracellular Signal-Regulated MAP Kinases ,Muscle, Skeletal ,Protein kinase C ,Genes, Dominant ,Endothelin-1 ,biology ,Myogenesis ,Gene Transfer Techniques ,General Medicine ,Receptor, Endothelin A ,Molecular biology ,Rats ,biology.protein ,Calcium ,Insulin Resistance ,Signal transduction ,Platelet-derived growth factor receptor ,Signal Transduction - Abstract
Aims Endothelin (ET) system plays a critical role in the development of insulin resistance and type 2 diabetes. In skeletal muscle, differentiation of myoblasts to myotubes is accompanied by the development of insulin sensitivity. Activation of extracellular signal-regulated kinase (ERK) 1/2 inhibits the differentiation of myoblasts, leading to insulin resistance. Although ET receptor (ETR) stimulation generally activates ERK1/2, the mechanism for ETR-mediated ERK1/2 activation in skeletal muscle is unknown. The purpose of this study was to determine the signal transduction pathway involved in ET-1-stimulated ERK1/2 phosphorylation in L6 myoblasts derived from rat skeletal muscle. Main methods Changes in phosphorylation levels of ERK1/2 following stimulation with ET-1 were analyzed by Western blot in L6 myoblasts. To inhibit receptor internalization, dominant-negative dynamin (K44A) was overexpressed in L6 myoblasts using adenovirus-mediated gene transfer. Key findings ET-1 induced phosphorylation of ERK1/2 in L6 myoblasts. The ERK1/2 phosphorylation was abolished by BQ123 (a selective ET type A receptor (ET A R) antagonist), YM-254890 (a G αq/11 protein inhibitor), and AG370 (a platelet-derived growth factor receptor (PDGFR) kinase inhibitor), while U-73122 (a phospholipase C (PLC) inhibitor) was less potent. The ERK1/2 phosphorylation was inhibited by overexpression of dominant-negative dynamin (K44A). These results suggest that ET A R stimulation induces ERK1/2 phosphorylation in L6 myoblasts through G q/11 protein-dependent, PLC-independent PDGFR transactivation which requires dynamin-dependent ET A R internalization. Significance Because activation of ERK1/2 is considered to inhibit differentiation of myoblasts with the development of insulin sensitivity, the ET A R-mediated PDGFR transactivation and subsequent ERK1/2 activation play an important role in ET-1-induced insulin resistance.
- Published
- 2014
6. Improved FRET Biosensor for the Measurement of BCR-ABL Activity in Chronic Myeloid Leukemia Cells
- Author
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Kosui Horiuchi, Xinxin Li, Shin-ya Nishide, Mari Fujioka, Asuka Nanbo, Yusuke Ohba, Prabha Nepal, Aya O. Satoh, Mika Horiguchi, Yoichiro Fujioka, Takeshi Kondo, and Takanori Teshima
- Subjects
0301 basic medicine ,Drug ,Physiology ,media_common.quotation_subject ,Fusion Proteins, bcr-abl ,Gene Expression ,Antineoplastic Agents ,Biosensing Techniques ,Cleavage (embryo) ,Transfection ,Biomarkers, Pharmacological ,03 medical and health sciences ,Leukemia, Myelogenous, Chronic, BCR-ABL Positive ,medicine ,Fluorescence Resonance Energy Transfer ,Humans ,Myeloid Cells ,Amino Acid Sequence ,Transgenes ,Phosphorylation ,Nuclear export signal ,Molecular Biology ,Protein Kinase Inhibitors ,media_common ,Adaptor Proteins, Signal Transducing ,Nuclear Export Signals ,Chemistry ,Myeloid leukemia ,Nuclear Proteins ,Cell Biology ,General Medicine ,medicine.disease ,CRKL ,Leukemia ,030104 developmental biology ,Amino Acid Substitution ,Immunology ,Cancer research ,K562 Cells ,Tyrosine kinase ,Plasmids - Abstract
Although the co-development of companion diagnostics with molecular targeted drugs is desirable, truly efficient diagnostics are limited to diseases in which chromosomal translocations or overt mutations are clearly correlated with drug efficacy. Moreover, even for such diseases, few methods are available to predict whether drug administration is effective for each individual patient whose disease is expected to respond to the drug(s). We have previously developed a biosensor based on the principle of Forster resonance energy transfer to measure the activity of the tyrosine kinase BCR-ABL and its response to drug treatment in patient-derived chronic myeloid leukemia cells. The biosensor harbors CrkL, one of the major substrates of BCR-ABL, and is therefore named Pickles after phosphorylation indicator of CrkL en substrate. The efficacy of this technique as a clinical test has been demonstrated, but the number of cells available for analysis is limited in a case-dependent manner, owing to the cleavage of the biosensor in patient-derived leukemia cells. Here, we describe an improved biosensor with an amino acid substitution and a nuclear export signal being introduced. Of the two predicted cleavage positions in CrkL, the mutations inhibited one cleavage completely and the other cleavage partially, thus collectively increasing the number of cells available for drug evaluation. This improved version of the biosensor holds promise in the future development of companion diagnostics to predict responses to tyrosine kinase inhibitors in patients with chronic myeloid leukemia.
- Published
- 2016
7. Different binding property of STIM1 and its novel splice variant STIM1L to Orai1, TRPC3, and TRPC6 channels
- Author
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Tsunehito Higashi, Takahiro Horinouchi, Prabha Nepal, Soichi Miwa, Hiroyuki Aoyagi, Tsunaki Higa, Takuya Harada, Mika Horiguchi, Koji Terada, Yosuke Mai, and Chizuru Hatate
- Subjects
inorganic chemicals ,Thapsigargin ,ORAI1 Protein ,Placenta ,Biophysics ,Biology ,Biochemistry ,Cerebral Ventricles ,TRPC6 ,Transient receptor potential channel ,chemistry.chemical_compound ,TRPC3 ,Pregnancy ,Human Umbilical Vein Endothelial Cells ,TRPC6 Cation Channel ,medicine ,Humans ,Protein Isoforms ,RNA, Messenger ,Stromal Interaction Molecule 1 ,Muscle, Skeletal ,Molecular Biology ,TRPC Cation Channels ,ORAI1 ,Endoplasmic reticulum ,Membrane Proteins ,Skeletal muscle ,STIM1 ,Cell Biology ,Neoplasm Proteins ,Cell biology ,HEK293 Cells ,medicine.anatomical_structure ,chemistry ,Calcium ,Female ,Calcium Channels ,HeLa Cells ,Protein Binding - Abstract
Stromal interaction molecule 1 (STIM1) is the endoplasmic reticulum (ER) Ca(2+) sensor to control ER Ca(2+) levels. A recent study has shown that STIM1L, a new splice variant of STIM1, is expressed in various tissues of rodent and in human skeletal muscle, and that the interaction of STIM1L with actin filament allows rapid activation of store-operated Ca(2+) entry (SOCE) mediated through Orai1 channels. Here, we characterize mRNA expression and function of human STIM1 and STIM1L, and compare their binding property to Orai1 functioning as store-operated Ca(2+) channels (SOCCs), and TRPC3 (transient receptor potential canonical 3) and TRPC6 channels functioning as endothelin type A receptor (ET(A)R)-operated Ca(2+) channels (ROCCs). Although mRNA for STIM1 was ubiquitously expressed in human tissues, STIM1L was detected only in skeletal muscle. STIM1L augmented thapsigargin- and endothelin-1-induced SOCE more strongly than STIM1 in human embryonic kidney 293 cells stably expressing ET(A)R, whereas, it tends to suppress ET(A)R-operated Ca(2+) entry (ROCE) via TRPC3 and TRPC6 more strongly than STIM1. Coimmunoprecipitation experiments have revealed that when compared with STIM1, STIM1L binds more abundantly to Orai1 and also to TRPC3 and TRPC6. These results suggest that the higher binding capacity of STIM1L to SOCCs and ROCCs plays an important role in the regulation of Ca(2+) signaling such as the augmentation of SOCE via Orai1 and the inhibition of ROCE via TRPC3 and TRPC6.
- Published
- 2012
8. A Sialylated Voltage-Dependent Ca2+ Channel Binds Hemagglutinin and Mediates Influenza A Virus Entry into Mammalian Cells
- Author
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Aya O. Satoh, Yusuke Ohba, Shin-ya Nishide, Yoichiro Fujioka, Mika Horiguchi, Yuko Sato, Izumi Kato, Sayaka Kashiwagi, Katsumi Maenaka, Sarad Paudel, Tadaaki Miyazaki, Hideki Hasegawa, Hideo Fukuhara, Kosui Horiuchi, Toyoyuki Ose, Tadaki Suzuki, Prabha Nepal, Jing Wang, Mari Fujioka, and Asuka Nanbo
- Subjects
0301 basic medicine ,Host cell surface ,Hemagglutinin (influenza) ,Biology ,medicine.disease_cause ,Microbiology ,Virus ,Cell biology ,Sialic acid ,03 medical and health sciences ,chemistry.chemical_compound ,030104 developmental biology ,Viral replication ,chemistry ,Virology ,Influenza A virus ,medicine ,biology.protein ,Parasitology ,Channel blocker ,Intracellular - Abstract
Influenza A virus (IAV) infection is initiated by the attachment of the viral glycoprotein hemagglutinin (HA) to sialic acid on the host cell surface. However, the sialic acid-containing receptor crucial for IAV infection has remained unidentified. Here, we show that HA binds to the voltage-dependent Ca2+ channel Cav1.2 to trigger intracellular Ca2+ oscillations and subsequent IAV entry and replication. IAV entry was inhibited by Ca2+ channel blockers (CCBs) or by knockdown of Cav1.2. The CCB diltiazem also inhibited virus replication in vivo. Reintroduction of wild-type but not the glycosylation-deficient mutants of Cav1.2 restored Ca2+ oscillations and virus infection in Cav1.2-depleted cells, demonstrating the significance of Cav1.2 sialylation. Taken together, we identify Cav1.2 as a sialylated host cell surface receptor that binds HA and is critical for IAV entry.
- Published
- 2018
9. A simple and rapid method for standard preparation of gas phase extract of cigarette smoke
- Author
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Soichi Miwa, Takuya Harada, Chizuru Hatate, Akimasa Hoshi, Mika Horiguchi, Koji Terada, Yosuke Mai, Yuji Kuge, Yoichi Noya, Takahiro Horinouchi, Prabha Nepal, and Tsunehito Higashi
- Subjects
Mts assay ,Time Factors ,Smoking habit ,Cytotoxicity ,viruses ,lcsh:Medicine ,Buffers ,Toxicology ,Gas phase ,Phosphates ,Tar (tobacco residue) ,Cell Line, Tumor ,Smoke ,Cigarette smoke ,Animals ,Humans ,lcsh:Science ,Reference standards ,Multidisciplinary ,Chromatography ,Cytotoxicity Assay ,Toxicity ,Chemistry ,Cytotoxins ,lcsh:R ,Temperature ,Reproducibility of Results ,Biology and Life Sciences ,Tobacco Products ,Reference Standards ,Hydrocarbons ,Rats ,lcsh:Q ,Gases ,Glass ,Filtration ,Research Article - Abstract
Cigarette smoke consists of tar and gas phase: the latter is toxicologically important because it can pass through lung alveolar epithelium to enter the circulation. Here we attempt to establish a standard method for preparation of gas phase extract of cigarette smoke (CSE). CSE was prepared by continuously sucking cigarette smoke through a Cambridge filter to remove tar, followed by bubbling it into phosphate-buffered saline (PBS). An increase in dry weight of the filter was defined as tar weight. Characteristically, concentrations of CSEs were represented as virtual tar concentrations, assuming that tar on the filter was dissolved in PBS. CSEs prepared from smaller numbers of cigarettes (original tar concentrations = 20 mg/ml), the curves were shifted rightward. Accordingly, the cytotoxic activity was detected in PBS of the second reservoir downstream of the first one with larger numbers of cigarettes. CSEs prepared from various cigarette brands showed comparable concentration-response curves for cytotoxicity. Two types of CSEs prepared by continuous and puff smoking protocols were similar regarding concentration-response curves for cytotoxicity, pharmacology of their cytotoxicity, and concentrations of cytotoxic compounds. These data show that concentrations of CSEs expressed by virtual tar concentrations can be a reference value to normalize their cytotoxicity, irrespective of numbers of combusted cigarettes, cigarette brands and smoking protocols, if original tar concentrations are
- Published
- 2014
10. Molecular mechanism for suppression of insulin signaling by endothelin-1 in skeletal muscle cells
- Author
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Soichi Miwa, Koji Terada, Yosuke Mai, Chizuru Hatate, Tsunehito Higashi, Akimasa Hoshi, Takuya Harada, Prabha Nepal, Mika Horiguchi, Takahiro Horinouchi, and Tsunaki Higa
- Subjects
biology ,Chemistry ,Biochemistry, Genetics and Molecular Biology(all) ,Skeletal muscle ,General Medicine ,Endothelin 1 ,General Biochemistry, Genetics and Molecular Biology ,Cell biology ,Insulin receptor ,Pharmacology, Toxicology and Pharmaceutics(all) ,medicine.anatomical_structure ,biology.protein ,Molecular mechanism ,medicine ,General Pharmacology, Toxicology and Pharmaceutics - Published
- 2013
- Full Text
- View/download PDF
11. Identification of stable cytotoxic factors in the gas phase extract of cigarette smoke and pharmacological characterization of their cytotoxicity
- Author
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Soichi Miwa, Takahiro Horinouchi, Prabha Nepal, Mika Horiguchi, Koh-ichi Seki, Hiroshi Asano, Yuji Kuge, Yoichi Noya, Chizuru Hatate, Koji Terada, Akimasa Hoshi, Tsunehito Higashi, and Yosuke Mai
- Subjects
Nicotine ,Cell Survival ,Cyclopentanes ,Toxicology ,Hydroxylamines ,Gas Chromatography-Mass Spectrometry ,Mass Spectrometry ,chemistry.chemical_compound ,Cell Line, Tumor ,Humans ,Propidium iodide ,Viability assay ,Cytotoxicity ,Protein kinase C ,Chromatography, High Pressure Liquid ,Protein Kinase C ,chemistry.chemical_classification ,Reactive oxygen species ,NADPH oxidase ,biology ,Cell-Free System ,Acrolein ,NADPH Oxidases ,Butanones ,Tars ,Biochemistry ,chemistry ,Apoptosis ,Data Interpretation, Statistical ,biology.protein ,Indicators and Reagents ,Tobacco Smoke Pollution - Abstract
Smoking is a major risk factor for atherosclerotic vascular diseases, but the mechanism for its genesis is unknown. We have recently shown that the gas phase of cigarette smoke (nicotine- and tar-free cigarette smoke extract; CSE) likely to reach the systemic circulation contains stable substances which cause cytotoxicity like plasma membrane damage and cell death in cultured cells, and also that the plasma membrane damage is caused through sequential activation of protein kinase C (PKC) and NADPH oxidase (NOX) and the resulting generation of reactive oxygen species (PKC/NOX-dependent mechanism), whereas cell death is caused through PKC/NOX-dependent and -independent mechanisms. To identify these stable substances, the CSE was prepared by passing the main-stream smoke of 10 cigarettes through a Cambridge glass fiber filter, trapping of the smoke in a vessel cooled at -80°C, and subsequent dissolution in 10ml of water. The CSE was fractionated into nine fractions using reversed-phase HPLC, and each fraction was screened for cytotoxicity in cultured cells, using propidium iodide uptake assay for cell membrane damage and MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium] reduction assay for cell viability. The cytotoxicity was positive in two of the nine fractions (Fr2 and Fr5). After extraction of the active fractions into dichloromethane, GC/MS analysis identified 2-cyclopenten-1-one (CPO) in Fr5 but none in Fr2. After derivatization of the active fractions with O-(2,3,4,5,6-pentafluorobenzyl) hydroxylamine hydrochloride, GC/MS analysis identified acrolein, acetone and propionaldehyde in Fr2, and methyl vinyl ketone (MVK) in Fr5. After 4-h incubation, authentic acrolein and MVK induced concentration-dependent cytotoxicity with EC50 values of 75.9±8.2 and 47.0±8.0μM (mean±SEM; n=3), respectively, whereas acetone, propionaldehyde and CPO were without effect. However, after 24-h incubation, CPO induced concentration-dependent cytotoxicity with an EC50 value of 264.0±16.9μM (n=3). The concentrations of acrolein, MVK and CPO in the CSE were 3368±334, 2429±123 and 392.9±31.8μM (n=4), respectively, which were higher than the cytotoxic concentrations. The cytotoxicity of acrolein and MVK consisted of plasma membrane damage and decreased cell viability: the plasma membrane damage was totally prevented by treatment with an inhibitor of PKC or NOX, whereas the decreased cell viability was only partially prevented by these inhibitors. The cytotoxicity of CPO consisted only of decreased cell viability, which was totally resistant to these inhibitors. These results show that acrolein and MVK are responsible for the acute cytotoxicity of the CSE through PKC/NOX-dependent and -independent mechanisms, whereas CPO is responsible for the delayed cytotoxicity of the CSE through a PKC/NOX-independent mechanism.
- Published
- 2013
12. Nicotine- and tar-free cigarette smoke extract induces cell injury via intracellular Ca2+-dependent subtype-specific protein kinase C activation
- Author
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Takuya Harada, Tsunehito Higashi, Yosuke Mai, Koji Terada, Prabha Nepal, Soichi Miwa, Takahiro Horinouchi, Mika Horiguchi, and Chizuru Hatate
- Subjects
Nicotine ,Protein Kinase C-alpha ,Protein Kinase C-epsilon ,Enzyme activator ,Tar (tobacco residue) ,Smoke ,parasitic diseases ,medicine ,Extracellular ,Tumor Cells, Cultured ,Animals ,Cell damage ,Protein kinase C ,Pharmacology ,NADPH oxidase ,biology ,Kinase ,Chemistry ,lcsh:RM1-950 ,Cell Membrane ,NADPH Oxidases ,Glioma ,Tobacco Products ,medicine.disease ,Molecular biology ,Tars ,Cell biology ,Rats ,Enzyme Activation ,lcsh:Therapeutics. Pharmacology ,biology.protein ,Molecular Medicine ,Calcium ,Reactive Oxygen Species ,Intracellular - Abstract
Nicotine- and tar-free cigarette smoke extract (CSE) is reported to induce cell damage via activation of protein kinase C (PKC) and NADPH oxidase (NOX) in rat C6 glioma cells. Here we determined PKC isozyme(s) activated by CSE and their activation mechanism. In C6 glioma cells, mRNAs for PKCα, PKCδ, PKCε, and PKCι were expressed. CSE triggered translocation of PKCα and PKCε to plasma membrane. CSE-induced cell damage and PKC translocation were inhibited by chelating intracellular Ca2+ but not extracellular Ca2+. These results suggest that CSE induces cell damage through intracellular Ca2+-dependent activation of PKCα and PKCε and subsequent NOX activation. Keywords:: cigarette smoke extract (CSE), protein kinase C (PKC), Ca2+
- Published
- 2012
13. Negative regulation of endothelin type A receptor-operated TRPC6 channel by adenylate cyclase-camp-protein kinase a signaling pathway
- Author
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Tsunaki Higa, Akimasa Hoshi, Prabha Nepal, Koji Terada, Takuya Harada, Yosuke Mai, Tsunehito Higashi, Takahiro Horinouchi, Chizuru Hatate, Soichi Miwa, and Mika Horiguchi
- Subjects
Endothelin receptor type A ,Biochemistry, Genetics and Molecular Biology(all) ,Chemistry ,Adenylate kinase ,General Medicine ,Cyclase ,General Biochemistry, Genetics and Molecular Biology ,TRPC6 ,Cell biology ,Pharmacology, Toxicology and Pharmaceutics(all) ,Channel (broadcasting) ,General Pharmacology, Toxicology and Pharmaceutics ,Protein kinase A signaling ,Endothelin receptor ,Receptor - Published
- 2013
14. Ubiquitin modification plays an important role in ET-1-dependent endothelin type B receptor trafficking
- Author
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Koji Terada, Prabha Nepal, Takahiro Horinouchi, Chizuru Hatate, Mika Horiguchi, Soichi Miwa, Akimasa Hoshi, Tsunehito Higashi, and Yosuke Mai
- Subjects
Pharmacology, Toxicology and Pharmaceutics(all) ,Ubiquitin ,biology ,Biochemistry, Genetics and Molecular Biology(all) ,Chemistry ,biology.protein ,General Medicine ,General Pharmacology, Toxicology and Pharmaceutics ,Receptor ,Endothelin receptor ,General Biochemistry, Genetics and Molecular Biology ,Cell biology - Full Text
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15. Endothelin-1 activates extracellular signal-regulated kinases 1 and 2 through transactivation of platelet-derived growth factor receptor in skeletal muscle cells
- Author
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Takuya Harada, Yosuke Mai, Tsunehito Higashi, Takahiro Horinouchi, Chizuru Hatate, Tsunaki Higa, Soichi Miwa, Mika Horiguchi, Koji Terada, Prabha Nepal, and Akimasa Hoshi
- Subjects
biology ,Biochemistry, Genetics and Molecular Biology(all) ,Chemistry ,Extracellular signal-regulated kinases ,digestive, oral, and skin physiology ,Skeletal muscle ,food and beverages ,General Medicine ,Endothelin 1 ,General Biochemistry, Genetics and Molecular Biology ,Cell biology ,Pharmacology, Toxicology and Pharmaceutics(all) ,Transactivation ,medicine.anatomical_structure ,Growth factor receptor ,embryonic structures ,biology.protein ,medicine ,cardiovascular system ,General Pharmacology, Toxicology and Pharmaceutics ,Platelet-derived growth factor receptor ,Insulin-like growth factor 1 receptor ,circulatory and respiratory physiology - Full Text
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16. Agonist-promoted Ubiquitination Differentially Regulates Receptor Trafficking of Endothelin Type A and Type B Receptors.
- Author
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Koji Terada, Takahiro Horinouchi, Yoichiro Fujioka, Tsunehito Higashi, Prabha Nepal, Mika Horiguchi, Sarita Karki, Chizuru Hatate, Akimasa Hoshi, Takuya Harada, Yosuke Mai, Yusuke Ohba, and Soichi Miwa
- Subjects
- *
G protein coupled receptors , *ENDOTHELINS , *PREPROENDOTHELIN , *LYSOSOMES , *CELL membranes , *LYSINE - Abstract
Two types of G protein-coupled receptors for endothelin-1 (ET-1), ET type A receptor (ETAR) and ETBR, closely resemble each other, but upon ET-1 stimulation, they follow totally different intracellular trafficking pathways; ETAR is recycled back to plasma membrane, whereas ETBR is targeted to lysosome for degradation. However, the mechanisms for such different fates are unknown. Here we demonstrated that ETBR but not ETAR was ubiquitinated on the cell surface following ET-1 stimulation and that ETBR was internalized and degraded in lysosome more rapidly than ETAR. The mutant ETBR (designated "5KR mutant") in which 5 lysine residues in the C-tail were substituted to arginine was not ubiquitinated, and its rates of internalization and degradation after ET-1 stimulation became slower, being comparable with those of ETAR. Confocal microscopic study showed that following ET-1 stimulation, ETAR and 5KR mutant of ETBR were co-localized mainly with Rab11, a marker of recycling endosome, whereas ETBR was co-localized with Rab7, a marker of late endosome/lysosome. In the 5KR mutant, ET-1-induced ERK phosphorylation and an increase in the intracellular Ca2+ concentration upon repetitive ET-1 stimulation were larger. A series of ETBR mutants (designated "4KR mutant"), in which either one of 5 arginine residues of the 5KR mutant was reverted to lysine, were normally ubiquitinated, internalized, and degraded, with ERK phosphorylation being normalized. These results demonstrate that agonist-induced ubiquitination at either lysine residue in the C-tail of ETBR but not ETAR switches intracellular trafficking from recycling to plasma membrane to targeting to lysosome, causing decreases in the cell surface level of ETBR and intracellular signaling. [ABSTRACT FROM AUTHOR]
- Published
- 2014
- Full Text
- View/download PDF
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