DOCSLIB.ORG

(TRP) Channels in Bladder Cancer Cells

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
(TRP) Channels in Bladder Cancer Cells J Physiol Sci (2014) 64:305–314 DOI 10.1007/s12576-014-0319-6 ORIGINAL PAPER Potential role of transient receptor potential (TRP) channels in bladder cancer cells Hideki Mizuno • Yoshiro Suzuki • Masaki Watanabe • Takaaki Sokabe • Tokunori Yamamoto • Ryohei Hattori • Momokazu Gotoh • Makoto Tominaga Received: 21 January 2014 / Accepted: 1 May 2014 / Published online: 22 May 2014 Ó The Physiological Society of Japan and Springer Japan 2014 Abstract Transient receptor potential (TRP) channels slowly, as compared to mock-transfected cells. Moreover, play important roles in thermal, chemical, and mechanical expression of dominant-negative TRPV2 significantly sensation in various tissues. In this study, we investigated decreased plasma membrane Ca2? permeability of MBT-2 the differences in urothelial TRP channels between normal cells as compared to that in mock-transfected cells. urothelial cells and bladder cancer cells. TRPV2 and Increases in the expression of TRPV2 mRNA, immunore- TRPM7 expression levels and TRPV2 activator-induced activity, and TRPV2 activator-induced intracellular Ca2? intracellular Ca2? increases were significantly higher, were also observed in T24 human bladder cancer cells. whereas TRPV4 expression and TRPV4 activator-induced These results suggested that TRPV2 and TRPM7 were intracellular Ca2? increases were significantly lower in functionally expressed in bladder cancer cells and served as mouse bladder cancer (MBT-2) cells compared to normal negative regulators of bladder cancer cell proliferation, mouse urothelial cells. The proliferation rate of MBT-2 most likely to prevent excess mechanical stresses. cells overexpressing dominant-negative TRPV2 was sig- nificantly increased. In contrast, treatment with TRPV2 Keywords Bladder cancer Á TRP channel Á Urothelial activators significantly decreased the proliferation rate. cells Á Mechanosensation TRPM7-overexpressing MBT-2 cells proliferated more Introduction Electronic supplementary material The online version of this article (doi:10.1007/s12576-014-0319-6) contains supplementary The transient receptor potential (TRP) superfamily of cation material, which is available to authorized users. channels plays critical roles in sensory physiology including H. Mizuno Á Y. Suzuki (&) Á M. Watanabe Á T. Sokabe Á thermal, chemical, and mechanical responses, in many types M. Tominaga (&) of cells. Recent studies have suggested that environmental Division of Cell Signaling, Okazaki Institute for Integrative factors, such as temperature, chemical exposure, pH, and Bioscience (National Institute for Physiological Sciences), National Institutes of Natural Sciences, 5-1 Higashiyama, mechanical stress, are sensed by specific TRP channels, Myodaiji, Okazaki 444-8787, Japan which may play a role in the regulation of proliferation, e-mail: [email protected] differentiation, apoptosis, and oncogenesis [1–4]. Changes M. Tominaga in the expression of TRPM1 in melanoma cells and TRPV6 e-mail: [email protected] and TRPM8 in prostate cancer cells have been reported [5– 8]. However, the physiological roles of these TRP channels H. Mizuno Á T. Yamamoto Á R. Hattori Á M. Gotoh Department of Urology, Nagoya University Graduate School of in cancer cells are still poorly understood. Medicine, 65 Tsurumai, Showa, Nagoya 466-8550, Japan Bladder cancer is the fourth most common cancer in men and has the eighth highest cancer-related mortality rate in Y. Suzuki Á M. Tominaga adult men [9]. Approximately 70–90 % of new cases are Department of Physiological Sciences, The Graduate University for Advanced Studies, 5-1, Higashiyama, Myodaiji, superficial tumors (stage Ta/T1/in situ). The standard treat- Okazaki 444-8787, Japan ment for these patients is transurethral resection. However, 123 306 J Physiol Sci (2014) 64:305–314 the tumor recurrence rate following transurethral resection of ‘‘everted ball bladder’’ was incubated with 0.05 % trypsin– bladder tumors is relatively high (30–60 %). Postoperative EDTA (Life Technologies, Carlsbad, CA, USA) for 30 min intravesical chemotherapy and immunotherapy have been on a shaker at 37 °C, and urothelial cells were harvested in used as adjuvant treatments in patients with a high risk of supplemented keratinocyte serum-free medium (KSFM; recurrence. However, these treatments only provide an 8 % Life Technologies) at 1 9 106 cells/mL. Drops of the cell reduction in recurrence in this patient population [9]. suspension were seeded on fibronectin (50 lg/mL)-coated Everaerts et al. [10] demonstrated the functional 60-mm dishes or cover slips (diameter 12 mm; Warner expression of TRPV4, TRPV2, and TRPM7 in mouse uro- Instruments Inc., Hamden, CT, USA). All experiments thelial cells. TRPV4, which is thought to be a sensor for with urothelial cells were performed after they had formed hypotonicity [11, 12], senses distension of the bladder clusters, approximately 72 h after the start of cultivation. urothelium; this signal is converted to an ATP signal in the micturition reflex pathway during urine storage [13]. A MBT-2 cells progressive decrease in the expression of the short splice- variant (s-TRPV2) accompanied by a marked increase in MBT-2 cells were derived from carcinogen (N-[4-(5-nitro-2- the expression of full-length TRPV2 (f-TRPV2) was found furyl)-2-thiazolyl]-formamide)-induced bladder tumors in in high-grade and advanced-stage urothelial carcinoma C3H/He mice. This cell line has been used as an animal model tissues [14]. In addition, TRPV2 has been reported to induce to evaluate the efficacy of antitumor drugs in bladder cancer apoptotic cell death in human T24 bladder cancer cells [15]. [16, 17]. MBT-2 cells (provided by Dr. Yamazaki, Kyoto However, the expression patterns and functions of TRP University, Japan) were maintained in RPMI-1640 medium channels in cancer are still not completely understood. supplemented with 10 % heat-inactivated fetal bovine serum, In this study, we sought to investigate the physiological 2mML-glutamine, nonessential amino acids (19), and 1 mM roles of TRP channels in bladder cancer cells by comparing sodium pyruvate at 37 °C in the presence of 5 % CO2. Cells the expression of TRP channels in normal mouse urothelial were passaged twice per week using 0.25 % trypsin and cells and MBT-2 bladder cancer cells. To elucidate the 0.02 % EDTA (Life Technologies). For MTS assays, 0.5 lg physiological implications of TRP channel expression, we of plasmid DNA containing dominant-negative TRPV2 [18, focused on the correlation between TRP channels and 19], rat TRPV2 (provided by Dr. David Julius, University of intracellular Ca2? homeostasis, which is one of the key California San Francisco, CA, USA), or human TRPM7 determinants of cancer cell proliferation. (provided by Dr. Yasuo Mori, Kyoto University, Japan) in OPTI-MEM (Life Technologies) were transfected into MBT- 2cells(29 104 cells/well in 24-well plates, Falcon) using Materials and methods Lipofectamine Plus Reagent (Life Technologies). After a 3.5- h incubation, the medium was changed to normal MBT-2 Chemicals medium, and cells were again incubated for 27 h at 37 °Cin the presence of 5 % CO2 before use. 2-Aminoethoxydiphenyl borate (2-APB), lysophosphati- dylcholine (LPC), GSK1016790A (GSK), and ionomycin T24 cells (Iono) were obtained from Sigma. The T24 cell line is a poorly differentiated human bladder Animals urothelial carcinoma cell line. T24 cells (obtained from the Human Science Research Resources Bank, Tokyo, Japan) All animal experiments were performed using 8-week-old were cultured in Eagle’s Minimum Essential Medium male wild-type (WT) C3H/HeN mice (CLEA Japan Inc., (EMEM; Wako, Osaka, Japan), containing 10 % heat- Tokyo, Japan). All procedures were conducted in accordance inactivated fetal bovine serum, 19 nonessential amino with the policies of the Institutional Animal Care and Use acids (Life Technologies), and 1 mM sodium pyruvate Committee, National Institute for Physiological Sciences. (Life Technologies) at 37 °C in the presence of 5 % CO2. Cells were passaged twice per week using 0.25 % trypsin Preparation of primary mouse urothelial cells and 0.02 % EDTA (Life Technologies). Whole bladders were taken from anesthetized mice, and Real-time polymerase chain reaction (PCR) of TRP urothelial cells were prepared using previously reported channels procedures [13], with slight modification. Briefly, the bladder was everted by pushing the dome downward Total RNA was isolated using a Sepasol G kit (Nakarai, through the bladder neck with a blunt 18-gauge needle. The Kyoto, Japan), and first-strand cDNA was synthesized 123 J Physiol Sci (2014) 64:305–314 307 using Superscript III reverse transcriptase (Life Technol- Measurement of plasma membrane Ca2? permeability ogies). Real-time PCR was performed with 10 lL SYBR Master Mix Reagent (Takara, Otsu, Japan), 0.25 lM To measure plasma membrane Ca2? permeability, we specific primer sets (for all TRPMs, TRPVs, TRPCs, and performed experiments using fura-2, as described previ- TRPA1; see Table S1), and 1 lL of the prepared cDNA. ously [20]. Dominant-negative TRPV2 was transfected into Real-time PCR analysis was performed using a StepOne MBT-2 cells together with Ds-Red plasmid (1/10 dilution), analyzer (Life Technologies). The temperature profile and Ca2? imaging was performed 24 h later using fura-2. consisted of 40 cycles of denaturation at 95 °C for 15 s, The initial Ca2? levels in the cells were measured, and cells annealing at 60 °C for 30 s, and elongation at 72 °C for were then treated with 2 mM EDTA solution without Ca2?.
Load more

Recommended publications
  • Transient Receptor Potential (TRP) Channels in Haematological Malignancies: an Update
    biomolecules Review Transient Receptor Potential (TRP) Channels in Haematological Malignancies: An Update Federica Maggi 1,2 , Maria Beatrice Morelli 2 , Massimo Nabissi 2 , Oliviero Marinelli 2 , Laura Zeppa 2, Cristina Aguzzi 2, Giorgio Santoni 2 and Consuelo Amantini 3,* 1 Department of Molecular Medicine, Sapienza University, 00185 Rome, Italy; [email protected] 2 Immunopathology Laboratory, School of Pharmacy, University of Camerino, 62032 Camerino, Italy; [email protected] (M.B.M.); [email protected] (M.N.); [email protected] (O.M.); [email protected] (L.Z.); [email protected] (C.A.); [email protected] (G.S.) 3 Immunopathology Laboratory, School of Biosciences and Veterinary Medicine, University of Camerino, 62032 Camerino, Italy * Correspondence: [email protected]; Tel.: +30-0737403312 Abstract: Transient receptor potential (TRP) channels are improving their importance in differ- ent cancers, becoming suitable as promising candidates for precision medicine. Their important contribution in calcium trafficking inside and outside cells is coming to light from many papers published so far. Encouraging results on the correlation between TRP and overall survival (OS) and progression-free survival (PFS) in cancer patients are available, and there are as many promising data from in vitro studies. For what concerns haematological malignancy, the role of TRPs is still not elucidated, and data regarding TRP channel expression have demonstrated great variability throughout blood cancer so far. Thus, the aim of this review is to highlight the most recent findings Citation: Maggi, F.; Morelli, M.B.; on TRP channels in leukaemia and lymphoma, demonstrating their important contribution in the Nabissi, M.; Marinelli, O.; Zeppa, L.; perspective of personalised therapies.
    [Show full text]
  • Snapshot: Mammalian TRP Channels David E
    SnapShot: Mammalian TRP Channels David E. Clapham HHMI, Children’s Hospital, Department of Neurobiology, Harvard Medical School, Boston, MA 02115, USA TRP Activators Inhibitors Putative Interacting Proteins Proposed Functions Activation potentiated by PLC pathways Gd, La TRPC4, TRPC5, calmodulin, TRPC3, Homodimer is a purported stretch-sensitive ion channel; form C1 TRPP1, IP3Rs, caveolin-1, PMCA heteromeric ion channels with TRPC4 or TRPC5 in neurons -/- Pheromone receptor mechanism? Calmodulin, IP3R3, Enkurin, TRPC6 TRPC2 mice respond abnormally to urine-based olfactory C2 cues; pheromone sensing 2+ Diacylglycerol, [Ca ]I, activation potentiated BTP2, flufenamate, Gd, La TRPC1, calmodulin, PLCβ, PLCγ, IP3R, Potential role in vasoregulation and airway regulation C3 by PLC pathways RyR, SERCA, caveolin-1, αSNAP, NCX1 La (100 µM), calmidazolium, activation [Ca2+] , 2-APB, niflumic acid, TRPC1, TRPC5, calmodulin, PLCβ, TRPC4-/- mice have abnormalities in endothelial-based vessel C4 i potentiated by PLC pathways DIDS, La (mM) NHERF1, IP3R permeability La (100 µM), activation potentiated by PLC 2-APB, flufenamate, La (mM) TRPC1, TRPC4, calmodulin, PLCβ, No phenotype yet reported in TRPC5-/- mice; potentially C5 pathways, nitric oxide NHERF1/2, ZO-1, IP3R regulates growth cones and neurite extension 2+ Diacylglycerol, [Ca ]I, 20-HETE, activation 2-APB, amiloride, Cd, La, Gd Calmodulin, TRPC3, TRPC7, FKBP12 Missense mutation in human focal segmental glomerulo- C6 potentiated by PLC pathways sclerosis (FSGS); abnormal vasoregulation in TRPC6-/-
    [Show full text]
  • TRPM7 Facilitates Cholinergic Vesicle Fusion with the Plasma Membrane
    TRPM7 facilitates cholinergic vesicle fusion with the plasma membrane Sebastian Brauchi*, Grigory Krapivinsky, Luba Krapivinsky, and David E. Clapham† Department of Cardiology, Howard Hughes Medical Institute, Children’s Hospital Boston, and Department of Neurobiology, Harvard Medical School, Enders Building 1309, 320 Longwood Avenue, Boston, MA 02115 Contributed by David E. Clapham, January 29, 2008 (sent for review December 3, 2007) TRPM7, of the transient receptor potential (TRP) family, is both an the inclusion of a zinc-finger domain (13). This kinase phos- ion channel and a kinase. Previously, we showed that TRPM7 is phorylates TRPM7 (14) and other substrates [e.g., annexin 1 (15) located in the membranes of acetylcholine (ACh)-secreting synaptic and myosin heavy chain (16)], but its physiological role is not vesicles of sympathetic neurons, forms a molecular complex with known. The pore of TRPM7 was proposed to be a major entry proteins of the vesicular fusion machinery, and is critical for mechanism for Mg2ϩ ions and, in this role, critical for cell stimulated neurotransmitter release. Here, we targeted pHluorin survival (17). TRPM7 is localized in the membrane of sympa- to small synaptic-like vesicles (SSLV) in PC12 cells and demonstrate thetic neuronal acetylcholine (ACh)-secreting synaptic vesicles that it can serve as a single-vesicle plasma membrane fusion as part of a molecular complex of synaptic vesicle-specific reporter. In PC12 cells, as in sympathetic neurons, TRPM7 is located proteins and its permeability is critical
    [Show full text]
  • The TRPM7 Ion Channel Functions in Cholinergic Synaptic Vesicles and Affects Transmitter Release
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector Neuron 52, 485–496, November 9, 2006 ª2006 Elsevier Inc. DOI 10.1016/j.neuron.2006.09.033 The TRPM7 Ion Channel Functions in Cholinergic Synaptic Vesicles and Affects Transmitter Release Grigory Krapivinsky,1 Sumiko Mochida,3 (TRPML channels) are presumed intracellular vesicular Luba Krapivinsky,1 Susan M. Cibulsky,1 channels (Di Palma et al., 2002; LaPlante et al., 2002), and David E. Clapham1,2,* but their function in vesicles is not well understood. 1 Howard Hughes Medical Institute Neurotransmitter vesicles release their cargoes by Cardiology fusion with the plasma membrane followed by diffusion Children’s Hospital Boston of their contents into the intercellular space. The fusion 1309 Enders Building process itself is not completely understood, but involves 320 Longwood Avenue a complex of proteins with specialized functions that al- 2 Department of Neurobiology low them to achieve close approximation of the vesicle Harvard Medical School to the plasma membrane and sense changes in [Ca2+] Boston, Massachusetts 02115 that trigger rapid fusion (Sudhof, 2004). Although neuro- 3 Department of Physiology transmitter release requires ion channels, ion channels Tokyo Medical University are best known for their function as the plasma mem- Tokyo 160-8402 brane mediators of the timing and entry of Ca2+ into Japan the cell, which in turn triggers vesicle fusion. Functional ion channels have been recorded in preparations of syn- aptic vesicles (Yakir and Rahamimoff, 1995; Kelly and Summary Woodbury, 1996), and a few well-established plasma membrane channels have been proposed to localize to A longstanding hypothesis is that ion channels are vesicles and regulate their ionic equilibrium (Ehrenstein present in the membranes of synaptic vesicles and et al., 1991; Grahammer et al., 2001), but their molecular might affect neurotransmitter release.
    [Show full text]
  • Free PDF Download
    European Review for Medical and Pharmacological Sciences 2019; 23: 3088-3095 The effects of TRPM2, TRPM6, TRPM7 and TRPM8 gene expression in hepatic ischemia reperfusion injury T. BILECIK1, F. KARATEKE2, H. ELKAN3, H. GOKCE4 1Department of Surgery, Istinye University, Faculty of Medicine, VM Mersin Medical Park Hospital, Mersin, Turkey 2Department of Surgery, VM Mersin Medical Park Hospital, Mersin, Turkey 3Department of Surgery, Sanlıurfa Training and Research Hospital, Sanliurfa, Turkey 4Department of Pathology, Inonu University, Faculty of Medicine, Malatya, Turkey Abstract. – OBJECTIVE: Mammalian transient Introduction receptor potential melastatin (TRPM) channels are a form of calcium channels and they trans- Ischemia is the lack of oxygen and nutrients port calcium and magnesium ions. TRPM has and the cause of mechanical obstruction in sev- eight subclasses including TRPM1-8. TRPM2, 1 TRPM6, TRPM7, TRPM8 are expressed especial- eral tissues . Hepatic ischemia and reperfusion is ly in the liver cell. Therefore, we aim to investi- a serious complication and cause of cell death in gate the effects of TRPM2, TRPM6, TRPM7, and liver tissue2. The resulting ischemic liver tissue TRPM8 gene expression and histopathologic injury increases free intracellular calcium. Intra- changes after treatment of verapamil in the he- cellular calcium has been defined as an important patic ischemia-reperfusion rat model. secondary molecular messenger ion, suggesting MATERIALS AND METHODS: Animals were randomly assigned to one or the other of the calcium’s effective role in cell injury during isch- following groups including sham (n=8) group, emia-reperfusion, when elevated from normal verapamil (calcium entry blocker) (n=8) group, concentrations. The high calcium concentration I/R group (n=8) and I/R- verapamil (n=8) group.
    [Show full text]
  • The Role of Transient Receptor Potential Channels in Metabolic Syndrome
    1989 Hypertens Res Vol.31 (2008) No.11 p.1989-1995 Review The Role of Transient Receptor Potential Channels in Metabolic Syndrome Daoyan LIU1), Zhiming ZHU1), and Martin TEPEL2) Metabolic syndrome is correlated with increased cardiovascular risk and characterized by several factors, including visceral obesity, hypertension, insulin resistance, and dyslipidemia. Several members of a large family of nonselective cation entry channels, e.g., transient receptor potential (TRP) canonical (TRPC), vanil- loid (TRPV), and melastatin (TRPM) channels, have been associated with the development of cardiovascular diseases. Thus, disruption of TRP channel expression or function may account for the observed increased cardiovascular risk in metabolic syndrome patients. TRPV1 regulates adipogenesis and inflammation in adi- pose tissues, whereas TRPC3, TRPC5, TRPC6, TRPV1, and TRPM7 are involved in vasoconstriction and reg- ulation of blood pressure. Other members of the TRP family are involved in regulation of insulin secretion, lipid composition, and atherosclerosis. Although there is no evidence that a single TRP channelopathy may be the cause of all metabolic syndrome characteristics, further studies will help to clarify the role of specific TRP channels involved in the metabolic syndrome. (Hypertens Res 2008; 31: 1989–1995) Key Words: metabolic syndrome, transient receptor potential channel, hypertension, cardiometabolic risk or diastolic blood pressure ≥85 mmHg; high fasting blood ≥ Metabolic Syndrome glucose 110 mg/dL (6.1 mmol/L); hypertriglyceridemia ≥150 mg/dL (1.7 mmol/L); or high-density lipoprotein cho- Metabolic syndrome is associated with several major risk lesterol <40 mg/dL (1.0 mmol/L) in men or <50 mg/dL (1.29 factors, including visceral obesity, hypertension, insulin mmol/L) in women.
    [Show full text]
  • TRP Channels of Intracellular Membranes
    JOURNAL OF NEUROCHEMISTRY | 2010 | 113 | 313–328 doi: 10.1111/j.1471-4159.2010.06626.x The Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, Michigan, USA Abstract potential (TRP) proteins are a superfamily (28 members in Ion channels are classically understood to regulate the flux of mammal) of Ca2+-permeable channels with diverse tissue ions across the plasma membrane in response to a variety of distribution, subcellular localization, and physiological func- environmental and intracellular cues. Ion channels serve a tions. Almost all mammalian TRP channels studied thus far, number of functions in intracellular membranes as well. These like their ancestor yeast TRP channel (TRPY1) that localizes channels may be temporarily localized to intracellular mem- to the vacuole compartment, are also (in addition to their branes as a function of their biosynthetic or secretory path- plasma membrane localization) found to be localized to ways, i.e., en route to their destination location. Intracellular intracellular membranes. Accumulated evidence suggests membrane ion channels may also be located in the endocytic that intracellularly-localized TRP channels actively participate pathways, either being recycled back to the plasma mem- in regulating membrane traffic, signal transduction, and brane or targeted to the lysosome for degradation. Several vesicular ion homeostasis. This review aims to provide a channels do participate in intracellular signal transduction; the summary of these recent works. The discussion will also be most well known example is the inositol 1,4,5-trisphosphate extended to the basic membrane and electrical properties of receptor (IP3R) in the endoplasmic reticulum. Some organellar the TRP-residing compartments.
    [Show full text]
  • TRPM Channels in Human Diseases
    cells Review TRPM Channels in Human Diseases 1,2, 1,2, 1,2 3 4,5 Ivanka Jimenez y, Yolanda Prado y, Felipe Marchant , Carolina Otero , Felipe Eltit , Claudio Cabello-Verrugio 1,6,7 , Oscar Cerda 2,8 and Felipe Simon 1,2,7,* 1 Faculty of Life Science, Universidad Andrés Bello, Santiago 8370186, Chile; [email protected] (I.J.); [email protected] (Y.P.); [email protected] (F.M.); [email protected] (C.C.-V.) 2 Millennium Nucleus of Ion Channel-Associated Diseases (MiNICAD), Universidad de Chile, Santiago 8380453, Chile; [email protected] 3 Faculty of Medicine, School of Chemistry and Pharmacy, Universidad Andrés Bello, Santiago 8370186, Chile; [email protected] 4 Vancouver Prostate Centre, Vancouver, BC V6Z 1Y6, Canada; [email protected] 5 Department of Urological Sciences, University of British Columbia, Vancouver, BC V6Z 1Y6, Canada 6 Center for the Development of Nanoscience and Nanotechnology (CEDENNA), Universidad de Santiago de Chile, Santiago 7560484, Chile 7 Millennium Institute on Immunology and Immunotherapy, Santiago 8370146, Chile 8 Program of Cellular and Molecular Biology, Institute of Biomedical Sciences (ICBM), Faculty of Medicine, Universidad de Chile, Santiago 8380453, Chile * Correspondence: [email protected] These authors contributed equally to this work. y Received: 4 November 2020; Accepted: 1 December 2020; Published: 4 December 2020 Abstract: The transient receptor potential melastatin (TRPM) subfamily belongs to the TRP cation channels family. Since the first cloning of TRPM1 in 1989, tremendous progress has been made in identifying novel members of the TRPM subfamily and their functions. The TRPM subfamily is composed of eight members consisting of four six-transmembrane domain subunits, resulting in homomeric or heteromeric channels.
    [Show full text]
  • TRPM7 Channels As a Bioassay of Internal and External Mg2+ a Thesis
    TRPM7 channels as a bioassay of internal and external Mg2+ A thesis submitted in partial fulfillment of the requirements for the degree of Master of Science by CHARLES T. LUU B.S., Ohio University, 2014 2019 Wright State University WRIGHT STATE UNIVERSITY GRADUATE SCHOOL September 25th, 2019 I HEREBY RECOMMEND THAT THE THESIS PREPARED UNDER MY SUPERVISION BY Charles T. Luu ENTITLED TRPM7 channels as a bioassay of internal and external Mg2+ BE ACCEPTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF Master of Science. _________________________ J. Ashot Kozak, Ph.D. Thesis Director __________________________ Eric Bennett, Ph.D Chair, Department of Neuroscience, Cell Biology and Physiology Committee on Final Examination: ________________________________ J.Ashot Kozak, Ph.D ________________________________ Mauricio Di Fulvio, Ph.D. ________________________________ Kathrin L. Engisch, Ph.D ________________________________ Barry Milligan, Ph.D. Interim Dean of the Graduate School ABSTRACT Luu, Charles.T. M.S., Department of Neuroscience, Cell Biology and Physiology, Wright State University, 2019. TRPM7 as a bioassay of internal and external Mg2+. Magnesium is an important divalent metal cation that is involved in numerous cellular functions. The details of cellular Mg2+ regulation, homeostasis and transport remain unclear. Magnesium transporter protein (MagT1) is a Mg2+ transporter and deficiency of this protein has been reported to lead to impaired Mg2+ influx and a decreased cytoplasmic [Mg2+]. Transient receptor potential melastatin 7 (TRPM7) is a ubiquitously expressed membrane protein containing a channel pore and a C-terminal alpha-type serine/threonine protein kinase domain. Importantly, TRPM7 channel is believed to conduct both Mg2+ and Ca2+. In the present study, we investigated if TRPM7 can be used as a bioassay of internal and external Mg2+ in Jurkat T cells.
    [Show full text]
  • Modulation of Transient Receptor Potential Melastatin 3 by Protons
    bioRxiv preprint doi: https://doi.org/10.1101/2020.10.08.331454; this version posted October 8, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC 4.0 International license. 1 Modulation of Transient receptor potential melastatin 3 by protons through its intracellular binding sites Md Zubayer Hossain Saad 1, 2, #, Liuruimin Xiang 1, 2, #, Yan-Shin Liao 3, Leah R. Reznikov 3, Jianyang Du1, 2, 4, * 1 Department of Anatomy and Neurobiology, University of Tennessee Health Science Center, Memphis, TN 38163, USA. 2 Department of Biological Sciences, University of Toledo, Toledo, OH 43606, USA. 3 Department of Physiological Sciences, University of Florida, Gainesville, FL 32610, USA. 4 Neuroscience Institute, University of Tennessee Health Science Center, Memphis, TN 38163, USA. # These authors contributed equally to this work Correspondence Jianyang Du, Tel: 901-448-3463, Email: [email protected] Running Title: Intracellular acidic pH inhibits TRPM3. bioRxiv preprint doi: https://doi.org/10.1101/2020.10.08.331454; this version posted October 8, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC 4.0 International license. 2 1 Abstract 2 Transient receptor potential melastatin 3 channel (TRPM3) is a calcium-permeable 3 nonselective cation channel that plays an important role in modulating glucose homeostasis 4 in the pancreatic beta cells.
    [Show full text]
  • Mapping TRPM7 Function by NS8593
    International Journal of Molecular Sciences Review Mapping TRPM7 Function by NS8593 Vladimir Chubanov * and Thomas Gudermann * Walther-Straub Institute of Pharmacology and Toxicology, Faculty of Medicine, Ludwig-Maximilians Universität München, 80336 Munich, Germany * Correspondence: [email protected] (V.C.); [email protected] (T.G.); Tel.: +49-89-2180-75740 (V.C.); +49-89-2180-75702 (T.G.) Received: 1 September 2020; Accepted: 21 September 2020; Published: 23 September 2020 Abstract: The transient receptor potential cation channel, subfamily M, member 7 (TRPM7) is a ubiquitously expressed membrane protein, which forms a channel linked to a cytosolic protein kinase. Genetic inactivation of TRPM7 in animal models uncovered the critical role of TRPM7 in early embryonic development, immune responses, and the organismal balance of Zn2+, Mg2+, and Ca2+. TRPM7 emerged as a new therapeutic target because malfunctions of TRPM7 have been associated with anoxic neuronal death, tissue fibrosis, tumour progression, and giant platelet disorder. Recently, several laboratories have identified pharmacological compounds allowing to modulate either channel or kinase activity of TRPM7. Among other small molecules, NS8593 has been defined as a potent negative gating regulator of the TRPM7 channel. Consequently, several groups applied NS8593 to investigate cellular pathways regulated by TRPM7. Here, we summarize the progress in this research area. In particular, two notable milestones have been reached in the assessment of TRPM7 druggability. Firstly, several laboratories demonstrated that NS8593 treatment reliably mirrors prominent phenotypes of cells manipulated by genetic inactivation of TRPM7. Secondly, it has been shown that NS8593 allows us to probe the therapeutic potential of TRPM7 in animal models of human diseases.
    [Show full text]
  • (TRP) Channels in Head-And-Neck Squamous Cell Carcinomas: Diagnostic, Prognostic, and Therapeutic Potentials
    International Journal of Molecular Sciences Review Transient Receptor Potential (TRP) Channels in Head-and-Neck Squamous Cell Carcinomas: Diagnostic, Prognostic, and Therapeutic Potentials 1, 2,3,4, , 5, 3,4,6, Fruzsina Kiss y, Krisztina Pohóczky * y , Arpad Szállási z and Zsuzsanna Helyes z 1 Somogy County Kaposi Mór Teaching Hospital, H-7400 Kaposvár, Hungary; [email protected] 2 Department of Pharmacology, Faculty of Pharmacy, University of Pécs, H-7624 Pécs, Hungary 3 Department of Pharmacology and Pharmacotherapy, Medical School, University of Pécs, H-7624 Pécs, Hungary; [email protected] 4 János Szentágothai Research Centre, Centre for Neuroscience, University of Pécs, H-7624 Pécs, Hungary 5 1st Department of Pathology and Experimental Cancer Research, Semmelweis University, H-1085 Budapest, Hungary; [email protected] 6 PharmInVivo Ltd., H-7629 Pécs, Hungary * Correspondence: [email protected]; Tel.: +36-72-536-001-35386 Fruzsina Kiss and Krisztina Pohóczky should be regarded as joint first authors. y Árpád Szállási and Zsuzsanna Helyes contributed equally to this paper. z Received: 15 July 2020; Accepted: 29 August 2020; Published: 2 September 2020 Abstract: Head-and-neck squamous cell carcinomas (HNSCC) remain a leading cause of cancer morbidity and mortality worldwide. This is a largely preventable disease with smoking, alcohol abuse, and human papilloma virus (HPV) being the main risk factors. Yet, many patients are diagnosed with advanced disease, and no survival improvement has been seen for oral SCC in the past decade. Clearly, new diagnostic and prognostic markers are needed for early diagnosis and to guide therapy. Gene expression studies implied the involvement of transient receptor potential (TRP) channels in the pathogenesis of HNSCC.
    [Show full text]