DOCSLIB.ORG

Distribution Profiles of Transient Receptor Potential Melastatin-Related and Vanilloid-Related Channels in Prostatic Tissue in Rat

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Distribution Profiles of Transient Receptor Potential Melastatin-Related and Vanilloid-Related Channels in Prostatic Tissue in Rat TRPM and TRPV in rat prostate DOI: 10.1111/j.1745-7262.2007.00291.x www.asiaandro.com .Original Article . Distribution profiles of transient receptor potential melastatin-related and vanilloid-related channels in prostatic tissue in rat Huai-Peng Wang*, Xiao-Yong Pu*, Xing-Huan Wang Department of Urology, Guangdong Provnicial People’s Hospital, Guangzhou 510080, China Abstract Aim: To investigate the expression and distribution of the members of the transient receptor potential (TRP) channel members of TRP melastatin (TRPM) and TRP vanilloid (TRPV) subfamilies in rat prostatic tissue. Methods: Pros- tate tissue was obtained from male Sprague-Dawley rats. Reverse transcription polymerase chain reaction (RT-PCR) and quantitative real-time polymerase chain reaction (PCR) were used to check the expression of all TRPM and TRPV channel members with specific primers. Immunohistochemistry staining for TRPM8 and TRPV1 were also per- formed in rat tissues. Results: TRPM2, TRPM3, TRPM4, TRPM6, TRPM7, TRPM8, TRPV2 and TRPV4 mRNA were detected in all rat prostatic tissues. Very weak signals for TRPM1, TRPV1 and TRPV3 were also detected. The mRNA of TRPM5, TRPV5 and TRPV6 were not detected in all RT-PCR experiments. Quantitative real-time RT-PCR showed that TRPM2, TRPM3, TRPM4, TRPM8, TRPV2 and TRPV4 were the most abundantly expressed TRPM and TRPV subtypes, respectively. Fluorescence immunohistochemistry indicated that TRPM8 and TRPV1 are highly expressed in both epithelial and smooth muscle cells. Conclusion: Our results demonstrate that mRNA or protein for TRPM1, TRPM2, TRPM3, TRPM4, TRPM6, TRPM7, TRPM8, TRPV1, TRPV2, TRPV3 and TRPV4 exist in rat prostatic tissue. The data presented here assists in elucidating the physiological function of TRPM and TRPV channels. (Asian J Androl 2007 Sep; 9: 634–640) Keywords: transient receptor potential channels; prostate; cation channels 1 Introduction Correspondence to: Dr Xing-Huan Wang, Department of Urology, Prostate cancer is the most commonly diagnosed Guangdong Provincial People’s Hospital, Guangzhou 510080, cancer in US men and its incidence rate is steadily rising China. in China [1]. It is possible to effectively treat organ- Tel: +86-20-8382-7812 Fax: +86-20-8382-7712 confined prostate cancer by radical prostatectomy and E-mail: [email protected] *The authors equally contribute to the study and preparation of radiation therapy [2]. Targeted-cryosurgical ablation of the manuscript. the prostate with androgen deprivation therapy improves Received 2006-11-19 Accepted 2007-04-04 the quality of life for high-risk prostate cancer patients. .634. © 2007, Asian Journal of Andrology, Shanghai Institute of Materia Medica,http://www.asiaandro.com; Chinese Academy of Sciences. All [email protected] rights reserved. Asian J Androl 2007; 9 (5): 634–640 However, there are very limited treatment options for tant subfamilies of the TRP superfamily, in rat prostate. metastatic prostate cancer. Therefore, it is of great im- Perhaps of the greatest interest is the observation that portance to find novel markers for diagnosing early stages some TRP channels appear to be expressed in a large of the disease and to closely monitor both progression spectrum of normal prostate tissue. Some might poten- and treatment of the disease, as well as to develop new tially be used as markers in prostate cancer diagnosis therapeutic approaches. To achieve this, we need to and for monitoring disease progression during treatment. screen candidate molecules for biomarkers and compare their expression in normal prostatic tissue and neoplastic 2 Materials and methods tissue. To date, there are very few prostate-specific markers available. The best-known and most well-char- 2.1 Rat tissue acterized markers for prostate cancer are prostate spe- Care of animals conformed to standards established cific antigen (PSA) [3] and prostate-specific membrane by the National Institute of Health of China. The antigen (PSMA) [4]. Each of these proteins has also Guangdong Provincial People’s Hospital Animal Care and become the target for novel immunotherapy approaches Use Committee approved all animal protocols. Three- to the treatment of the disease [5, 6]. month-old Sprague-Dawley rats were killed with pento- The transient receptor potential (TRP) channels, have barbital sodium (130 mg/kg i.p.). They were exsangui- not yet been systematically studied in prostate even nated, and the prostate were isolated and removed quickly. though some subfamilies have been explored. The TRP The prostate was cleaned free of connective tissue. protein superfamily consists of a diverse group of cation channels that bear structural similarities to Drosophila 2.2 Reverse transcription reaction (PCR) TRP. Based on sequence homology, the TRP superfa- The prostate tissue was obtained and maintained at mily can be divided into three major subfamilies of ca- –80ºC until processing for mRNA expression analysis. nonical (TRPC), melastatin-related (TRPM) and vanilloid- Total RNA was extracted using TRIzol reagent related (TRPV) channels and into four more distant sub- (Invitrogen, Carlsbad, CA, USA) from the prostate tis- families of polycystin (TRPP), mucolipin (TRPML), sue using the standard procedures. In a 20-μL reaction ankyrin (TRPA), and no mechanoreceptor potential C or 1 μg of total RNA was resuspended in dihexadecylphospha- NOMPC (TRPN)-related channels/proteins. Of all the tidylcholine (DEPC)-treated water. The reverse transcrip- TRP superfamily, only the TRPC subfamily is a store- tion (RT) reaction was performed using an RT system pro- operated channel, whereas others appear to be activated cedure (Promega, Madison, USA). Briefly, 5 mmol/L by production of diacylglycerol or regulated through an MgCl2, 1 × RT buffer (10 mmol/L tris-HCl, 50 mmol/L KCl exocytotic mechanism. Many members of TRPV func- and 0.1% Triton X-100), 1 mmol/L deoxynucleoside triph- tion in sensory physiology and respond to heat, changes osphate (dNTP) mixture (equal amounts of deoxyadenosine in osmolarity, odorants and mechanical stimuli. Two triphosphate [dATP], deoxycytidine triphosphate [dCTP], members of the TRPM family function in sensory per- deoxyguanosine triphosphate [dGTP] and deoxythymidine ception and three TRPM proteins are chanzymes, which triphosphate [dTTP]) (Sigma-Aldrich), 1 μg/μL recombi- contain C-terminal enzyme domains. As for TRPN and nant RNase (Jingmei Biotech, Shenzhen, China) ribonu- TRPA, the proteins are characterized by many ankyrin clease inhibitor, 15 U avian myeloblastosis virus (dAMV) repeats. TRPN proteins function in mechanotransduction, RT (high concentration) and 0.5 µg oligodeoxythymidine whereas TRPA1 is activated by noxious cold and is also (oligo DT) 15 primers were added to the RNA mixture. required for auditory response. In addition to these five The reaction was done at 37ºC for 5 min and 42ºC for closely related TRP subfamilies, which comprise the 1 h, followed by 10 min of heating at 95ºC to destroy the Group 1 TRP, members of the two Group 2 TRP enzyme and RNA. subfamilies, TRPP and TRPML, are distantly related to the Group 1 TRP. Mutations in the found members of 2.3 Conventional PCR amplification these latter subfamilies are responsible for human dis- Sense and anti-sense PCR primers specific to the eases [7]. TRPM and TRPV channels were designed as Yang et al. In the present study, we systematically analyzed the [8]. RT reaction (1 µL) was amplified in a 20-µL reac- expression of TRPM and TRPV channels, two impor- tion containing 1 × PCR buffer, 0.5 unit Taq (Promega, Tel: +86-21-5492-2824; Fax: +86-21-5492-2825; Shanghai, China .635. TRPM and TRPV in rat prostate Madision, NJ, USA) DNA polymerase, 0.25 mmol/L Indirect immunohistochemistry staining was per- dNTP mixture (equal amounts of dATP, dCTP, dGTP formed using the conventional procedure. The primary and dTTP), 0.2 mmol/L sense and antisense primers, antibodies were the same as those used for the Western and 0.25 µL dimethyl sulfoxide (DMSO). Cycle param- blot. Briefly, paraffin section slides were de-paraffinated eters consisted of a 30-s denaturing step at 94ºC, 45-s with 100% xylenes for three times, each time for 5 min, annealing step at 56ºC and 90-s extension step at 72ºC then the sections were rehydrated with 100%, 85% and with 35 cycles per amplification. This was followed by 70% alcohol. Slides were placed in PBS for 5 min, and a final extension at 72ºC for 10 min and the products antigen retrieval was done by covering the sections with were then stored at 4ºC. PCR products were electro- 0.02 mol/L citrate buffer, pH 6.0, and heating them in a mi- phoresed on 1.8% agarose gel (Jingmei Biotech, crowave oven for 3 min. After cooling for 3 min, addi- Shenzhen, China), stained with 0.5 µg/mL ethidium, and tional buffer was added and slides were reheated. This visualized and photographed on an ultraviolet transillu- process was repeated until four cycles of heating and minator bromide (SIM International, Los Angeles, CA, cooling had been performed. Slides were cooled down USA). Parallel reactions were run for each RNA sample to room temperature for 20 min, rinsed with PBS and in the absence of Superscript III to ascertain that there blocked with 8% normal goat serum in PBS for 45 min. was no genomic DNA contamination. The sections were incubated overnight at 4ºC with pri- mary antibody containing 3% serum. After washing three 2.4 Quantitative real-time PCR times with PBS/0.05% Tween-20 (PBST), sections were Gene-specific real-time PCR primers were designed incubated for 1 h at room temperature with Cy3-conju- based upon the published TRPM or TRPV sequences in gated goat anti-rabbit antibody (1:800, Jackson Genbank to obtain predicted PCR products of 100–150 Immunolab, West Grove, USA). Sections were thor- bases. At least one primer of each set was designed to oughly washed with PBST, and then counterstained with span exon-exon junctions in order to minimize the possi- the nuclear dye Hoechest 32258 (1:1 200, Sigma; St.
Load more

Recommended publications
  • The Role of Transient Receptor Potential Cation Channels in Ca2þ Signaling
    Downloaded from http://cshperspectives.cshlp.org/ on October 7, 2021 - Published by Cold Spring Harbor Laboratory Press The Role of Transient Receptor Potential Cation Channels in Ca2þ Signaling Maarten Gees, Barbara Colsoul, and Bernd Nilius KU Leuven, Department of Molecular Cell Biology, Laboratory Ion Channel Research, Campus Gasthuisberg, Herestraat 49, bus 802, Leuven, Belgium Correspondence: [email protected] The 28 mammalian members of the super-family of transient receptor potential (TRP) channels are cation channels, mostly permeable to both monovalent and divalent cations, and can be subdivided into six main subfamilies: the TRPC (canonical), TRPV (vanilloid), TRPM (melastatin), TRPP (polycystin), TRPML (mucolipin), and the TRPA (ankyrin) groups. TRP channels are widely expressed in a large number of different tissues and cell types, and their biological roles appear to be equally diverse. In general, considered as poly- modal cell sensors, they play a much more diverse role than anticipated. Functionally, TRP channels, when activated, cause cell depolarization, which may trigger a plethora of voltage-dependent ion channels. Upon stimulation, Ca2þ permeable TRP channels 2þ 2þ 2þ generate changes in the intracellular Ca concentration, [Ca ]i,byCa entry via the plasma membrane. However, more and more evidence is arising that TRP channels are also located in intracellular organelles and serve as intracellular Ca2þ release channels. This review focuses on three major tasks of TRP channels: (1) the function of TRP channels as Ca2þ entry channels; (2) the electrogenic actions of TRPs; and (3) TRPs as Ca2þ release channels in intracellular organelles. ransient receptor potential (TRP) channels choanoflagellates, yeast, and fungi are primary Tconstitute a large and functionally versatile chemo-, thermo-, or mechanosensors (Cai 2008; family of cation-conducting channel proteins, Wheeler and Brownlee 2008; Chang et al.
    [Show full text]
  • The Structural Basis for an On–Off Switch Controlling Gβγ-Mediated Inhibition of TRPM3 Channels
    The structural basis for an on–off switch controlling Gβγ-mediated inhibition of TRPM3 channels Marc Behrendta,b,1,2, Fabian Grussc,1,3, Raissa Enzerotha,b, Sandeep Demblaa,b, Siyuan Zhaod, Pierre-Antoine Crassousd, Florian Mohra, Mieke Nysc, Nikolaos Lourose, Rodrigo Gallardoe,4, Valentina Zorzinif,5, Doris Wagnera, Anastassios Economou (Αναστάσιoς Οικoνόμoυ)f, Frederic Rousseaue, Joost Schymkowitze, Stephan E. Philippg, Tibor Rohacsd, Chris Ulensc,6, and Johannes Oberwinklera,b,6 aInstitut für Physiologie und Pathophysiologie, Philipps-Universität Marburg, 35037 Marburg, Germany;bCenter for Mind, Brain and Behavior (CMBB), Philipps-Universität Marburg and Justus-Liebig-Universität Giessen, 35032 Marburg, Germany; cLaboratory of Structural Neurobiology, Department of Cellular and Molecular Medicine, KU Leuven, 3000 Leuven, Belgium; dDepartment of Pharmacology, Physiology and Neuroscience, Rutgers New Jersey Medical School, Newark, NJ 07103; eSwitch Laboratory, VIB Center for Brain and Disease Research, Department of Cellular and Molecular Medicine, KU Leuven, 3000 Leuven, Belgium; fLaboratory of Molecular Bacteriology, Department of Microbiology and Immunology, Rega Institute for Medical Research, KU Leuven, 3000 Leuven, Belgium; and gExperimentelle und Klinische Pharmakologie und Toxikologie, Universität des Saarlandes, 66421 Homburg, Germany Edited by László Csanády, Semmelweis University, Budapest, Hungary, and accepted by Editorial Board Member David E. Clapham August 27, 2020 (received for review February 13, 2020) TRPM3 channels play important roles in the detection of noxious also subject to inhibition by activated μORsorotherGPCRs(6, heat and in inflammatory thermal hyperalgesia. The activity of 24–28), a process that has been shown to take place in peripheral these ion channels in somatosensory neurons is tightly regulated by endings of nociceptive neurons since locally applied μOR or μ βγ -opioid receptors through the signaling of G proteins, thereby GABA receptor agonists inhibit TRPM3-dependent pain (24–26).
    [Show full text]
  • The TRPP2-Dependent Channel of Renal Primary Cilia Also Requires TRPM3
    The TRPP2-dependent channel of renal primary cilia also requires TRPM3 Steven J. Kleene1, Brian J. Siroky2, Julio A. Landero-Figueroa3, Bradley P. Dixon4, Nolan W. Pachciarz2, Lu Lu2, and Nancy K. Kleene1 1Department of Pharmacology and Systems Physiology, University of Cincinnati, Cincinnati, Ohio; 2Division of Nephrology and Hypertension, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio; 3Department of Chemistry, University of Cincinnati, Cincinnati, Ohio; 4Renal Section, Department of Pediatrics, University of Colorado School of Medicine, Aurora, Colorado IntroductIon Primary cilia of renal epithelial cells express several members of the transient receptor potential (TRP) class of cation-conducting channel, including TRPC1, TRPM3, TRPM4, TRPP2, and TRPV4. Some cases of autosomal dominant polycystic kidney disease (ADPKD) are caused by defects in TRPP2 (also called polycystin-2, PC2, or PKD2). A large-conductance, TRPP2- dependent channel in renal cilia has been well described, but it is not known whether this channel includes any other protein subunits. Methods To study this question, we investigated the pharmacology of the TRPP2-dependent channel through electrical recordings from the cilia of mIMCD-3 cells, a murine cell line of renal epithelial origin, results The pharmacology was found to match that of TRPM3 channels. The ciliary TRPP2-dependent channel is known to be activated by depolarization and/or increasing cytoplasmic Ca2+. This activation was greatly enhanced by external pregnenolone sulfate, an agonist of TRPM3 channels. Pregnenolone sulfate did not change the current-voltage relation of the channel. CIM0216, another TRPM3 agonist, modestly increased the activity of the ciliary channels. The channels were effectively blocked by isosakuranetin, a specific inhibitor of TRPM3 channels.
    [Show full text]
  • Disease-Associated Mutations in TRPM3 Render the Channel Overactive Via Two Distinct Mechanisms
    bioRxiv preprint doi: https://doi.org/10.1101/2020.04.20.052167; this version posted April 22, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Disease-associated mutations in TRPM3 render the channel overactive via two distinct mechanisms Siyuan Zhao, Yevgen Yudin, Tibor Rohacs Department of Pharmacology, Physiology and Neuroscience, New Jersey Medical School, Rutgers University, Newark, NJ ABSTRACT Transient Receptor Potential Melastatin 3 (TRPM3) is a Ca2+ permeable non-selective cation channel activated by heat and chemical agonists such as pregnenolone sulfate and CIM0216. TRPM3 mutations in humans were recently reported to be associated with intellectual disability and epilepsy; the functional effects of those mutations however were not reported. Here we show that both disease-associated mutations of TRPM3 render the channel overactive, but likely via different mechanisms. The Val to Met substitution in the S4-S5 loop induced a larger increase in basal activity and agonist sensitivity at room temperature than the Pro to Gln substitution in the extracellular segment of S6. In contrast, heat activation was increased more by the S6 mutant than by the S4-S5 segment mutant. Both mutants were inhibited by the TRPM3 antagonist primidone, suggesting a potential therapeutic intervention to treat this disease. INTRODUCTION Transient Receptor Potential Melastatin 3 (TRPM3) is a Ca2+ permeable, non-selective cation channel activated by heat (Vriens et al., 2011) and chemical activators such as the neurosteroid pregnenolone sulfate (PregS) (Wagner et al., 2008) and the synthetic compound CIM0216 (Held et al., 2015).
    [Show full text]
  • Recessive Mutations of the Gene TRPM1 Abrogate on Bipolar Cell Function and Cause Complete Congenital Stationary Night Blindness in Humans
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector REPORT Recessive Mutations of the Gene TRPM1 Abrogate ON Bipolar Cell Function and Cause Complete Congenital Stationary Night Blindness in Humans Zheng Li,1 Panagiotis I. Sergouniotis,1 Michel Michaelides,1,2 Donna S. Mackay,1 Genevieve A. Wright,2 Sophie Devery,2 Anthony T. Moore,1,2 Graham E. Holder,1,2 Anthony G. Robson,1,2 and Andrew R. Webster1,2,* Complete congenital stationary night blindness (cCSNB) is associated with loss of function of rod and cone ON bipolar cells in the mammalian retina. In humans, mutations in NYX and GRM6 have been shown to cause the condition. Through the analysis of a consan- guineous family and screening of nine additional pedigrees, we have identified three families with recessive mutations in the gene TRPM1 encoding transient receptor potential cation channel, subfamily M, member 1, also known as melastatin. A number of other variants of unknown significance were found. All patients had myopia, reduced central vision, nystagmus, and electroretinographic evidence of ON bipolar cell dysfunction. None had abnormalities of skin pigmentation, although other skin conditions were reported. RNA derived from human retina and skin was analyzed and alternate 50 exons were determined. The most 50 exon is likely to harbor an initiation codon, and the protein sequence is highly conserved across vertebrate species. These findings suggest an important role of this specific cation channel for the normal function of ON bipolar cells in the human retina. Congenital stationary night blindness (CSNB) is a group of of the gene encoding transient receptor potential cation genetically determined, nondegenerative disorders of the channel, subfamily M, member 1 (TRPM1 [MIM *603576]) retina associated with lifelong deficient vision in the dark has been discovered in the skin and retina of horses homo- and often nystagmus and myopia.
    [Show full text]
  • The TRPM4 Channel Inhibitor 9-Phenanthrol
    W&M ScholarWorks Arts & Sciences Articles Arts and Sciences 2014 The TRPM4 channel inhibitor 9-phenanthrol R. Guinamard T. Hof C. A. Del Negro College of William and Mary Follow this and additional works at: https://scholarworks.wm.edu/aspubs Recommended Citation Guinamard, R., Hof, T., & Del Negro, C. A. (2014). The TRPM4 channel inhibitor 9‐phenanthrol. British Journal of Pharmacology, 171(7), 1600-1613. This Article is brought to you for free and open access by the Arts and Sciences at W&M ScholarWorks. It has been accepted for inclusion in Arts & Sciences Articles by an authorized administrator of W&M ScholarWorks. For more information, please contact [email protected]. British Journal of DOI:10.1111/bph.12582 www.brjpharmacol.org BJP Pharmacology REVIEW Correspondence Romain Guinamard, Groupe Signalisation, Electrophysiologie et Imagerie des Lésions The TRPM4 channel d’Ischémie/Reperfusion Myocardique, EA4650, Université de Caen Basse Normandie, inhibitor 9-phenanthrol Sciences D, Esplande de la Paix, CS 14032, 14032 Caen Cedex 5, R Guinamard1,2, T Hof1 and C A Del Negro2 France. E-mail: [email protected] ---------------------------------------------------------------- 1EA 4650, Groupe Signalisation, Electrophysiologie et Imagerie des Lésions d’Ischémie-Reperfusion Myocardique, UCBN, Normandie Université, Caen, France, 2Department Keywords 9-phenanthrol; TRPM4; of Applied Science, The College of William and Mary, Williamsburg, VA, USA calcium-activated non-selective cation channel; cardioprotection; NSCCa ---------------------------------------------------------------- Received 1 November 2013 Revised 17 December 2013 Accepted 8 January 2014 The phenanthrene-derivative 9-phenanthrol is a recently identified inhibitor of the transient receptor potential melastatin (TRPM) 4 channel, a Ca2+-activated non-selective cation channel whose mechanism of action remains to be determined.
    [Show full text]
  • Investigational Drugs in Early Phase Clinical Trials Targeting Thermotransient Receptor Potential (Thermotrp) Channels
    Expert Opinion on Investigational Drugs ISSN: (Print) (Online) Journal homepage: https://www.tandfonline.com/loi/ieid20 Investigational drugs in early phase clinical trials targeting thermotransient receptor potential (thermoTRP) channels Asia Fernández-Carvajal , Rosario González-Muñiz , Gregorio Fernández- Ballester & Antonio Ferrer-Montiel To cite this article: Asia Fernández-Carvajal , Rosario González-Muñiz , Gregorio Fernández- Ballester & Antonio Ferrer-Montiel (2020): Investigational drugs in early phase clinical trials targeting thermotransient receptor potential (thermoTRP) channels, Expert Opinion on Investigational Drugs, DOI: 10.1080/13543784.2020.1825680 To link to this article: https://doi.org/10.1080/13543784.2020.1825680 Published online: 29 Sep 2020. Submit your article to this journal Article views: 31 View related articles View Crossmark data Full Terms & Conditions of access and use can be found at https://www.tandfonline.com/action/journalInformation?journalCode=ieid20 EXPERT OPINION ON INVESTIGATIONAL DRUGS https://doi.org/10.1080/13543784.2020.1825680 REVIEW Investigational drugs in early phase clinical trials targeting thermotransient receptor potential (thermoTRP) channels Asia Fernández-Carvajala, Rosario González-Muñizb, Gregorio Fernández-Ballestera and Antonio Ferrer-Montiela aInstituto De Investigación, Desarrollo E Innovación En Biotecnología Sanitaria De Elche (Idibe), Universitas Miguel Hernández, Alicante, Spain; bInstituto De Química Médica, CSIC, Madrid, Spain ABSTRACT ARTICLE HISTORY Introduction: Thermo transient receptor potential (thermoTRP) channels are some of the most inten­ Received 15 June 2020 sely pursued therapeutic targets of the past decade. They are considered promising targets of numer­ Accepted 15 September ous diseases including chronic pain and cancer. Modulators of these proteins, in particular TRPV1-4, 2020 TRPM8 and TRPA1, have reached clinical development, but none has been approved for clinical practice KEYWORDS yet.
    [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]
  • Involvement of TRPC4 and 5 Channels in Persistent Firing in Hippocampal CA1 Pyramidal Cells
    cells Article Involvement of TRPC4 and 5 Channels in Persistent Firing in Hippocampal CA1 Pyramidal Cells Alberto Arboit 1,2,3, Antonio Reboreda 1,4 and Motoharu Yoshida 1,3,4,5,* 1 German Center for Neurodegenerative Diseases (DZNE), 39120 Magdeburg, Germany; [email protected] (A.A.); [email protected] (A.R.) 2 Otto-von-Guericke University, 39120 Magdeburg, Germany 3 Faculty of Psychology, Ruhr University Bochum (RUB), Universitätsstraße 150, 44801 Bochum, Germany 4 Leibniz Institute for Neurobiology (LIN), 39118 Magdeburg, Germany 5 Center for Behavioral Brain Sciences (CBBS), 39106 Magdeburg, Germany * Correspondence: [email protected] Received: 1 December 2019; Accepted: 1 February 2020; Published: 5 February 2020 Abstract: Persistent neural activity has been observed in vivo during working memory tasks, and supports short-term (up to tens of seconds) retention of information. While synaptic and intrinsic cellular mechanisms of persistent firing have been proposed, underlying cellular mechanisms are not yet fully understood. In vitro experiments have shown that individual neurons in the hippocampus and other working memory related areas support persistent firing through intrinsic cellular mechanisms that involve the transient receptor potential canonical (TRPC) channels. Recent behavioral studies demonstrating the involvement of TRPC channels on working memory make the hypothesis that TRPC driven persistent firing supports working memory a very attractive one. However, this view has been challenged by recent findings that persistent firing in vitro is unchanged in TRPC knock out (KO) mice. To assess the involvement of TRPC channels further, we tested novel and highly specific TRPC channel blockers in cholinergically induced persistent firing in mice CA1 pyramidal cells for the first time.
    [Show full text]
  • Heteromeric TRP Channels in Lung Inflammation
    cells Review Heteromeric TRP Channels in Lung Inflammation Meryam Zergane 1, Wolfgang M. Kuebler 1,2,3,4,5,* and Laura Michalick 1,2 1 Institute of Physiology, Charité—Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 10117 Berlin, Germany; [email protected] (M.Z.); [email protected] (L.M.) 2 German Centre for Cardiovascular Research (DZHK), 10785 Berlin, Germany 3 German Center for Lung Research (DZL), 35392 Gießen, Germany 4 The Keenan Research Centre for Biomedical Science, St. Michael’s Hospital, Toronto, ON M5B 1W8, Canada 5 Department of Surgery and Physiology, University of Toronto, Toronto, ON M5S 1A8, Canada * Correspondence: [email protected] Abstract: Activation of Transient Receptor Potential (TRP) channels can disrupt endothelial bar- rier function, as their mediated Ca2+ influx activates the CaM (calmodulin)/MLCK (myosin light chain kinase)-signaling pathway, and thereby rearranges the cytoskeleton, increases endothelial permeability and thus can facilitate activation of inflammatory cells and formation of pulmonary edema. Interestingly, TRP channel subunits can build heterotetramers, whereas heteromeric TRPC1/4, TRPC3/6 and TRPV1/4 are expressed in the lung endothelium and could be targeted as a protec- tive strategy to reduce endothelial permeability in pulmonary inflammation. An update on TRP heteromers and their role in lung inflammation will be provided with this review. Keywords: heteromeric TRP assemblies; pulmonary inflammation; endothelial permeability; TRPC3/6; TRPV1/4; TRPC1/4 Citation: Zergane, M.; Kuebler, W.M.; Michalick, L. Heteromeric TRP Channels in Lung Inflammation. Cells 1. Introduction 2021, 10, 1654. https://doi.org Pulmonary microvascular endothelial cells are a key constituent of the blood air bar- /10.3390/cells10071654 rier that has to be extremely thin (<1 µm) to allow for rapid and efficient alveolo-capillary gas exchange.
    [Show full text]
  • Ca Signaling in Cardiac Fibroblasts and Fibrosis-Associated Heart
    Journal of Cardiovascular Development and Disease Review Ca2+ Signaling in Cardiac Fibroblasts and Fibrosis-Associated Heart Diseases Jianlin Feng 1, Maria K. Armillei 1, Albert S. Yu 1, Bruce T. Liang 1, Loren W. Runnels 2,* and Lixia Yue 1,* 1 Calhoun Cardiology Center, Department of Cell Biology, University of Connecticut Health Center, Farmington, CT 06030, USA; [email protected] (J.F.); [email protected] (M.K.A.); [email protected] (A.S.Y.); [email protected] (B.T.L.) 2 Department of Pharmacology, Rutgers, Robert Wood Johnson Medical School, Piscataway, NJ 08854, USA * Correspondence: [email protected] (L.W.R.); [email protected] (L.Y.) Received: 11 August 2019; Accepted: 18 September 2019; Published: 23 September 2019 Abstract: Cardiac fibrosis is the excessive deposition of extracellular matrix proteins by cardiac fibroblasts and myofibroblasts, and is a hallmark feature of most heart diseases, including arrhythmia, hypertrophy, and heart failure. This maladaptive process occurs in response to a variety of stimuli, including myocardial injury, inflammation, and mechanical overload. There are multiple signaling pathways and various cell types that influence the fibrogenesis cascade. Fibroblasts and myofibroblasts are central effectors. Although it is clear that Ca2+ signaling plays a vital role in this pathological process, what contributes to Ca2+ signaling in fibroblasts and myofibroblasts is still not wholly understood, chiefly because of the large and diverse number of receptors, transporters, and ion channels that influence intracellular Ca2+ signaling. Intracellular Ca2+ signals are generated by Ca2+ release from intracellular Ca2+ stores and by Ca2+ entry through a multitude of Ca2+-permeable ion channels in the plasma membrane.
    [Show full text]
  • Ion Channels 3 1
    r r r Cell Signalling Biology Michael J. Berridge Module 3 Ion Channels 3 1 Module 3 Ion Channels Synopsis Ion channels have two main signalling functions: either they can generate second messengers or they can function as effectors by responding to such messengers. Their role in signal generation is mainly centred on the Ca2 + signalling pathway, which has a large number of Ca2+ entry channels and internal Ca2+ release channels, both of which contribute to the generation of Ca2 + signals. Ion channels are also important effectors in that they mediate the action of different intracellular signalling pathways. There are a large number of K+ channels and many of these function in different + aspects of cell signalling. The voltage-dependent K (KV) channels regulate membrane potential and + excitability. The inward rectifier K (Kir) channel family has a number of important groups of channels + + such as the G protein-gated inward rectifier K (GIRK) channels and the ATP-sensitive K (KATP) + + channels. The two-pore domain K (K2P) channels are responsible for the large background K current. Some of the actions of Ca2 + are carried out by Ca2+-sensitive K+ channels and Ca2+-sensitive Cl − channels. The latter are members of a large group of chloride channels and transporters with multiple functions. There is a large family of ATP-binding cassette (ABC) transporters some of which have a signalling role in that they extrude signalling components from the cell. One of the ABC transporters is the cystic − − fibrosis transmembrane conductance regulator (CFTR) that conducts anions (Cl and HCO3 )and contributes to the osmotic gradient for the parallel flow of water in various transporting epithelia.
    [Show full text]