Tyrosine Kinase Pathways, Smooth Muscle Function and Inos Induction

Total Page:16

File Type:pdf, Size:1020Kb

Tyrosine Kinase Pathways, Smooth Muscle Function and Inos Induction University of Calgary PRISM: University of Calgary's Digital Repository Graduate Studies Legacy Theses 1997 Tyrosine kinase pathways, smooth muscle function and iNOS induction Zheng, Xilong Zheng, X. (1997). Tyrosine kinase pathways, smooth muscle function and iNOS induction (Unpublished doctoral thesis). University of Calgary, Calgary, AB. doi:10.11575/PRISM/17957 http://hdl.handle.net/1880/26938 doctoral thesis University of Calgary graduate students retain copyright ownership and moral rights for their thesis. You may use this material in any way that is permitted by the Copyright Act or through licensing that has been assigned to the document. For uses that are not allowable under copyright legislation or licensing, you are required to seek permission. Downloaded from PRISM: https://prism.ucalgary.ca THE UNIVERSITY OF CALGARY TYROSINE KINASE PATHWAYS, SMOOTEI MUSCLE FUNCTION AND iNOS INDUCTION by Xiiong Zheng A DISSERTATION SUBMKTED TO TEE FACULTY OF GRADUATE STUDiES IN PARTIAL FULE*ILMENT OF THE REQUIREMENTS FOR TEE DEGREE OF DOCTOR OF PEILOSOPHY DEPARTMENT OF MEDICAL SCIENCE CALGARY, ALBERTA JUNE, 1997 0Xüong Zheng 1997 IYPIIOMI uurary OIUIIUU I~UUI IOUUI I~JU 1*1 oiCanada du Canada Acquisitions and Acquisitions et Bibliographie Services services bibliographiques 395 Weilington Street 395, nie Wellington Ottawa ON KlA ON4 Ottawa ON K1A ON4 Canada Canada The author has granted a non- L'auteur a accordé une Licence non exclusive licence ailowing the exclusive permettant a la National Library of Canada to Bibliothèque nationafe du Canada de reproduce, loan, distriiute or sell reproduire, prêter, distniuer ou copies of this thesis in microfonn, vendre des copies de cette thése sous paper or electronic formats. la forme de rnicrofiche/fh, de reproduction sur papier ou sur format électronique. The author retains ownership of the L'auteur conserve la propriété du copyright in this thesis. Neither the droit d'auteur qui protège cette thèse. thesis nor substantial extracts fiom it Ni Ia thèse ni des extraits substantiels may be printed or othewise de celle-ci ne doivent être imprimés reproduced without the author's ou autrement reproduits sans son permission. autorisation- ABSTRACT The role(s) of tyrosine kinase (TYK) pathways in both a conirade response and the induction of induciile nitric oxide synthase (iNOS)have been studied in vasniIar and gastric smooth muscle (SM) tissue. In guinea pig (GP) gastric Iongituduial muscle (LM), contractions caused by epidermal growth factor-urogastrone (EGF), ethanol and a thrombin receptor activating peptide (PAR@) were found to share signalhg pathway enzymes, including TM(, diacylglycerol lipase and cyclooxygenase. The LM responses to EGF and PAR,AP also involved MAP kinase-kinase (MEX), phosphatidyhositol3-kinase and protein kinase C, whereas the ethanol response did not. Ethanol PAR,AP and EGF- induced responses were al1 dependent on extracellular Ca2'; the EGF and PAR,AP responses involved a voltage-operated Ca2+charnel whereas the ethanol response involved a receptor-operated channel. Neither the ethano1 nor the PAR,AP contractile response were due to transactivation of the EGF receptor. In the organ bath, WOS was induced spontaneously in rat aorta (RA) and rat gastric circular muscle (RGCM)tissue, but not in RGLM.in the RA, but not in the CM, both a tyrosine kinase inhibitor (AG213) and a tyrosine phosphatase inhibitor (vanadate) blocked fùnctional iNOS induction. Whereas vanadate blocked both iNOS mRNA induction and the induction of BOS enzyme activity, AG2 13 blocked only the appearance of enzyme activity, as assesseci by an L-arginine relaxation response. In the RA but not in RGCM, iNOS induction was dependent on NF-kB activity; in the RGCM but not in the RA tissue, iNOS induction depended on the synthesis of a cycloheximide-sensitive factor. In the RGCM tissue, tNOS induction occurred in the macrophage-related cens, but not, as in the RA tissue, in the iii smooth muscle element. It was concluded that, depending on the smooth muscle phenotype, TYKs and tyrosine phosphatases may play a role in smooth muscle bction, both in tems of regulating the aaite contractile response and in tems of modulating enzyme induction as typified by NOS. There are no words in the world that can be found for me to express my appreciation to my supervisor, Dr. Morley D. Hollenberg, to whom 1 am deeply indebted. Without Dr. Hollenberg's generous support and bis excellent supervision, the work presented in this thesis wuld not have been accomplished. 1would iike to thank Drs. David L. Severson and Chris R. Triggle, my supervisory cornmittee members, for their wonderfiil suggestions and encouragement during my study. Dr. Keith A Sharkey is highly appreciated for his help with the immunohistochemical studies in this thesis. 1 would also Iike to thank Dr. Sultan Ahmad for his heIp with the molecular biological studies and Ms. Winnie Ho for her technical help with irnmunoùistochemical studies. Dr. Mike Walsh and Dr. Roger Loutzenhiser who were examiners in my candidacy examination are also highly appreciated. The input of Drs. Steven L. Pelech and Keith A Sharkey as the extemal examiners of this thesis is also gratefdiy acknowIedged. Partial sdary support that provided for the completion of this thesis work was provided by Wiam H. Davies Medical Research Scholarship. DEDICATION TO MY MOTHER TO MY WIFE, YU AND MY DAUGHTER JIE TABLE OF CONTENTS APPROVALPAGE ................................................... u.. DEDICATION ...................................................... vi TABLEOFCONTENTS .............................................vi LISTOFTABLES ............................................ xiv LISTOFFIGURES .................................................. xv LISTOFABBREVIATIONS ......................................... xxi CHAPTER ONE: INTRODUCTION ..................................... 1 1.1.0veMew .................................................. 1 1.2. Mechanisrns of smooth muscle contraction and relaxation .............. 5 1 -2.1. Caz' dependent contractile machinery ...................... 5 1.2.2. Regdation of intraceMar fiee Ca2' ....................... 6 1.2.3. Myosin regdation ..................................... 9 1.2.4. Thin filament regdation of smooth muscle ................. 12 1.2.5. Protein kinaseC ..................................... 14 1.3. The role of tyrosine kinase in signal transduction .................... 15 1.3.1.Introduction ........................................ 15 1.3 .2. Tyrosine kinase receptors .............................. 16 1-3 -3 . Activation of c-Src ................................... 18 1-3 .4 . The mitogen activated protein (MAP) kinase cascade ......... 19 1 -3.5 . Phosphatidylinositol3- (PI 3-j kinase ..................... 22 1 .3.6.G protein coupled receptors: Introduction .................. 23 1.3.7. Gpy and the MAP kinase cascade ........................ 27 1 -3.7.1. Gpy-mediated Ras dependent MAP kinase pathway ... 28 1.3.7.2. Tyrosine kinase pathways and MAPK ..............29 1 -3.7.3 . Tyrosine kinases as candidates in the activation of MAPK by G protein coupled receptor .................... 30 1-3 -7 .4 . ûther signaling moIdes ....................... 35 1.3.8. G protein Gq and the activation of MAP kinase ............. 37 1.3.9. Tyrosine kinase pathways and smooth muscle fbnction ........ 38 1.4. Signal transduction pathways and the induction of nitric oxide synthase (mas) ................................................. 41 1 .4.1. Nïm~oxide synthesis ................................. 41 viii 1.4.2. Sigaalling byN0 .................................... 42 1 .4.3. NOS isoforms and distribution .......................... 45 1.4.4. Structure of NOS .................................... 50 1.4.5. Signal transduction and ïNOS induction ...................51 1.4.5.2. NOS gene ..................................51 1.4.5.2.ïNQS mRNA stabüity .......................... 52 1 .4.5 .3 . Functional regulation of 240s ................... 52 1.4.5.4. Signalling for iNOS gene transcription .............53 1.4.5.5. Tyrosine kinase pathway and BOS induction ........ 56 1.5 . PreIiminary observations and rationale for the work described in the thesis ...................................................... 58 1.5.1.Background ........................................ 58 1.5.2. Preliary expenmeuts ............................... 60 1.5.3. Main goak of the thesis ............................... 61 CHAPTER TWO: MATERIALS AND METHODS ........................ 63 2.1. Materials and reagents ........................................ 63 2.2. Bioassay procedures ......................................... 64 2.2.1. Gastric smooth muscle preparation ....................... 64 2.2.2. Rat aorta preparation ................................. 67 2.3. Western blot analysis ......................................... 68 2.4. Preparation of tissue RN& reverse transcriptase-polymerase chah reaction analysis and nuckotide quencing ............................ 69 2.5. L-arginïne-mediateci relaxation assay ............................. 70 2.6. Immunohistochemistry ....................................... 71 CHAPTER THREE: SIGNALLING PATBWAYS FOR THE CONTRACTLE ACTION OF ETHANOL, EGF AND THROMBIN RECEPTOR ACTIVATINGPEPTIDE ........................................ 72 3.1.Introduction ............................................... 72 3.2. Ethanol-induced contractile response ............................. 75 3.2.1. Effts of tyrosine kinase inhibitors, indomethacin and inhibitors of nerve-released
Recommended publications
  • Studio Delle Vie Di Trasduzione Del Segnale Inositide-Dipendente Nelle Sindromi Mielodisplastiche
    UNIVERSITA' DEGLI STUDI DI BOLOGNA Scuola di Dottorato in Scienze Mediche e Chirurgiche Cliniche Dottorato di Ricerca in Scienze Morfologiche Umane e Molecolari Settore Disciplinare BIO/16 Dipartimento di Scienze Anatomiche Umane e Fisiopatologia dell’Apparato Locomotore STUDIO DELLE VIE DI TRASDUZIONE DEL SEGNALE INOSITIDE-DIPENDENTE NELLE SINDROMI MIELODISPLASTICHE Tesi di Dottorato Tutore: Presentata da: CHIAR.MO PROF. LUCIO COCCO DOTT.SSA MATILDE YUNG FOLLO XIX Ciclo Anno Accademico 2005/2006 INDICE Introduzione 3 1. Sindromi Mielodisplastiche (MDS) 4 1.1.Trattamento delle MDS: 5’-azacitidina 8 2. Signalling Inositide-Dipendente: Fosfolipasi cβ1 (PI-PLCβ1) 10 2.1 Struttura del Gene della PI-PLCβ1 11 2.2 Struttura Proteica della PI-PLCβ1 12 3. Asse di Attivazione Fosfoinositide-3-Chinasi (PI3K)/Akt 14 3.1 Isoforme di Akt 16 3.2. Ruolo di Akt nei Disordini Ematopoietici 18 3.3. Ruolo di Akt nei Meccanismi Apoptotici 19 3.4. Ruolo di Akt nella Progressione attraverso il Ciclo Cellulare 20 4. Target Molecolari a Valle di Akt: mTOR, 4E-BP1e p70S6K 21 Scopo della Ricerca 23 Materiali e Metodi 25 1. Colture Cellulari in vitro 26 2. Caratteristiche dei Pazienti 26 3. Separazione delle Cellule Mononucleate 26 4. Ibridazione Fluorescente in Situ (FISH) 27 5. Estrazione del DNA ed Analisi Mutazionale 28 6. Estrazione dell’RNA e Sintesi del cDNA 28 7. Real-Time PCR 28 8. Analisi Immunocitochimica 29 9. Separazione delle Cellule CD33 31 10. Analisi Citofluorimetrica per la Quantificazione dell’Apoptosi 31 11. Analisi Citofluorimetrica per l’Analisi del Fenotipo 32 12. Separazione delle Cellule CD34 33 13.
    [Show full text]
  • Cephalic Sensory Cell Types Provides Insight Into Joint Photo
    RESEARCH ARTICLE Characterization of cephalic and non- cephalic sensory cell types provides insight into joint photo- and mechanoreceptor evolution Roger Revilla-i-Domingo1,2,3, Vinoth Babu Veedin Rajan1,2, Monika Waldherr1,2, Gu¨ nther Prohaczka1,2, Hugo Musset1,2, Lukas Orel1,2, Elliot Gerrard4, Moritz Smolka1,2,5, Alexander Stockinger1,2,3, Matthias Farlik6,7, Robert J Lucas4, Florian Raible1,2,3*, Kristin Tessmar-Raible1,2* 1Max Perutz Labs, University of Vienna, Vienna BioCenter, Vienna, Austria; 2Research Platform “Rhythms of Life”, University of Vienna, Vienna BioCenter, Vienna, Austria; 3Research Platform "Single-Cell Regulation of Stem Cells", University of Vienna, Vienna BioCenter, Vienna, Austria; 4Division of Neuroscience & Experimental Psychology, University of Manchester, Manchester, United Kingdom; 5Center for Integrative Bioinformatics Vienna, Max Perutz Labs, University of Vienna and Medical University of Vienna, Vienna, Austria; 6CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria; 7Department of Dermatology, Medical University of Vienna, Vienna, Austria Abstract Rhabdomeric opsins (r-opsins) are light sensors in cephalic eye photoreceptors, but also function in additional sensory organs. This has prompted questions on the evolutionary relationship of these cell types, and if ancient r-opsins were non-photosensory. A molecular profiling approach in the marine bristleworm Platynereis dumerilii revealed shared and distinct *For correspondence: features of cephalic and non-cephalic r-opsin1-expressing cells. Non-cephalic cells possess a full set [email protected] (FR); of phototransduction components, but also a mechanosensory signature. Prompted by the latter, [email protected] (KT-R) we investigated Platynereis putative mechanotransducer and found that nompc and pkd2.1 co- Competing interest: See expressed with r-opsin1 in TRE cells by HCR RNA-FISH.
    [Show full text]
  • Role of Endothelin-1 in the Gastrointestinal Tract of Horses In
    Louisiana State University LSU Digital Commons LSU Doctoral Dissertations Graduate School 2003 Role of endothelin-1 in the gastrointestinal tract of horses in health and disease Ramaswamy Monickarasi Chidambaram Louisiana State University and Agricultural and Mechanical College Follow this and additional works at: https://digitalcommons.lsu.edu/gradschool_dissertations Part of the Veterinary Medicine Commons Recommended Citation Chidambaram, Ramaswamy Monickarasi, "Role of endothelin-1 in the gastrointestinal tract of horses in health and disease" (2003). LSU Doctoral Dissertations. 1717. https://digitalcommons.lsu.edu/gradschool_dissertations/1717 This Dissertation is brought to you for free and open access by the Graduate School at LSU Digital Commons. It has been accepted for inclusion in LSU Doctoral Dissertations by an authorized graduate school editor of LSU Digital Commons. For more information, please [email protected]. ROLE OF ENDOTHELIN-1 IN THE GASTROINTESTINAL TRACT OF HORSES IN HEALTH AND DISEASE A Dissertation Submitted to the Graduate Faculty of the Louisiana State University and Agricultural and Mechanical College in partial fulfillment of the requirements for the degree of Doctor of Philosophy The Interdepartmental Program in Veterinary Medical Sciences through the Department of Comparative Biomedical Sciences By Ramaswamy M. Chidambaram BVSc, Madras Veterinary College, India, 1996 MSc, Atlantic Veterinary College, Canada, 2000 May, 2003 Dedicated to my parents, Dr. S. Chidambaram Pillai and Mrs. R. Monickarasi, and my siblings for their inspiration and support toward my pursuit of higher knowledge ii ACKNOWLEDGEMENTS I express my sincere thanks and heartfelt gratitude to my mentor Dr. Rustin Moore and Dr. Changaram Venugopal, for their involvement and personal help offered toward the completion of my dissertation.
    [Show full text]
  • On the Mechanism of Task Channel Inhibition by G-Protein Coupled Receptors
    Aus dem Institut für Physiologie und Pathophysiologie (Geschäftsführender Direktor: Prof. Dr. Dr. Jürgen Daut) des Fachbereichs Medizin der Philipps-Universität Marburg ON THE MECHANISM OF TASK CHANNEL INHIBITION BY G-PROTEIN COUPLED RECEPTORS Inaugural-Dissertation zur Erlangung des Doktorgrades der gesamten Humanmedizin, dem Fachbereich Medizin der Philipps-Universität Marburg vorgelegt von Moritz Lindner aus Witten Marburg, 2012 Angenommen vom Fachbereich Medizin der Philipps-Universität Marburg am: 13. Dezember 2012 Gedruckt mit Genehmigung des Fachbereichs. Dekan: Prof. Dr. Matthias Rothmund Referent: Prof. Dr. Dominik Oliver 1. Korreferent: Prof. Dr. Timothy David Plant To my dear parents Abbreviations AMP-PCP β,γ-Methyleneadenosine-5′-triphosphate ATP Adenosine-tri-phosphate BAPTA 2,2′-(Ethylenedioxy)dianiline-N,N,N′,N′-tetraacetic acid CF-Inp54 Fusion construct of a CFP with the FKBP domain from the FK506 binding protein and the yeast Inp54p Phosphatase CFP Cyan fluorescent protein CHO Chinese Hamster Ovary Ci-VSP Ciona intestinalis voltage-sensitive phosphatase DAG Diacylglycerol DNA Deoxyribonucleic acid EGTA Ethylenglycol-bis(aminoethylether)-N,N,N′N′-tetraacetic acid Et-1 Endothelin-1 Ex-0 Standard extracellular solution GFP Green-fluorescent protein GPCR G-protein coupled receptor GqPCR Gq-protein coupled receptor Gqα Q-type alpha-subunit of the G-protein GTP Guanosine-tri-phosphate HEPES 4-(2-Hydroxyethyl)piperazine-1-ethanesulfonic acid ICS Intracellular solution Ins(1,4,5)P3 Inositol-1,4,5-tris-phosphate K2P Two-pore-domain
    [Show full text]
  • Pharmacological Characterisation of the Fatty Acid Receptors GPR120
    Pharmacological characterisation of the fatty acid receptors GPR120 and FFA1 Sarah-Jane Watson, BSc. Thesis submitted to the University of Nottingham for the degree of Doctor of Philosophy NOVEMBER 2013 Abstract In recent years, two G protein coupled receptors have been de-orphanised which respond to long chain free fatty acids (FFAs), and so are able to mediate the signalling of these important nutrient molecules. FFA1 (GPR40) is predominantly expressed in pancreatic -cells, while the expression profile of GPR120 includes gut endocrine cells and adipose tissue. These distributions, together with the potential of both receptors to stimulate insulin and incretin hormone secretion, singled them out as potential drug targets for type 2 diabetes and obesity. The aim of this thesis was to evaluate the pharmacology of these receptors and their signalling properties, including the development of fluorescent FFA receptor ligands to evaluate agonist binding using imaging techniques. GPR120 has been identified to exist as two splice isoforms in humans, differing by a short insertion in the third intracellular loop, but no full isoform specific characterisation of receptor signalling and trafficking had been undertaken. This work therefore studied the GPR120S and GPR120L isoforms in terms of both G protein dependent and arrestin dependent signalling, and trafficking. It was found that the long GPR120L isoform exhibited reduced G protein signalling, but similar -arrestin recruitment and lysosomal intracellular trafficking profiles as GPR120S. Potentially, expression of the long GPR120 isoform provides a mechanism to direct signalling to the -arrestin pathway, for example to produce anti-inflammatory effects in macrophages. As the expression profile of GPR120 overlaps with that of FFA1, for example in i colonic endocrine cells, a series of constrained GPR120 homo-dimers and GPR120:FFA1 heterodimers were created using irreversible bimolecular fluorescence complementation, and the potential for novel pharmacology was investigated by monitoring dimer internalisation.
    [Show full text]
  • ERK1/2: an Integrator of Signals That Alters Cardiac Homeostasis and Growth
    biology Review ERK1/2: An Integrator of Signals That Alters Cardiac Homeostasis and Growth Christopher J. Gilbert †, Jacob Z. Longenecker † and Federica Accornero * Department of Physiology and Cell Biology, Dorothy M. Davis Heart and Lung Research Institute, The Ohio State University, Columbus, OH 43210, USA; [email protected] (C.J.G.); [email protected] (J.Z.L.) * Correspondence: [email protected] † Equal contribution. Simple Summary: Understanding how cardiac cells respond to external stimuli is essential for devel- oping interventions that mitigate pathologies of the heart. Therefore, in this review, we summarize critical knowledge related to a key molecular pathway that mediates cellular responses. Abstract: Integration of cellular responses to extracellular cues is essential for cell survival and adaptation to stress. Extracellular signal-regulated kinase (ERK) 1 and 2 serve an evolutionarily conserved role for intracellular signal transduction that proved critical for cardiomyocyte homeostasis and cardiac stress responses. Considering the importance of ERK1/2 in the heart, understanding how these kinases operate in both normal and disease states is critical. Here, we review the complexity of upstream and downstream signals that govern ERK1/2-dependent regulation of cardiac structure and function. Particular emphasis is given to cardiomyocyte hypertrophy as an outcome of ERK1/2 activation regulation in the heart. Citation: Gilbert, C.J.; Longenecker, J.Z.; Accornero, F. ERK1/2: An Keywords: ERK; extracellular matrix; heart; cardiac hypertrophy Integrator of Signals That Alters Cardiac Homeostasis and Growth. Biology 2021, 10, 346. https:// doi.org/10.3390/biology10040346 1. Introduction Within the heart, cardiomyocytes represent the primary working cell responsible Academic Editor: for contraction.
    [Show full text]
  • Supporting Information
    Supporting Information Tong et al. 10.1073/pnas.0904571106 SI Results Arrestin-Beta. The arrestin-beta gene sequence of E. scolopes predicted a protein longer than that of L. pealei. Both visual and Characteristics of the E. scolopes Sequences Characterized in nonvisual arrestins have been identified in both invertebrates this Study and vertebrates, but thus far, only the visual system-specific R-Opsin. Sequences in the eye and light organ were identical to arrestin has been identified in cephalopods (7). The derived each other and to the sequence for E. scolopes visual rhodopsin amino acid sequence of the E. scolopes light-organ arrestin that had already been reported (1), a finding demonstrating that contains the 5 fingerprint regions that are characteristic of these the same isoform of this protein probably occurs in both the eyes proteins; in these regions, the overall identity is 89%. In addition, and light organs of this squid species. Comparisons of the 5 polar core residues, which are present in the bovine and L. sequence with that of rhodopsins of other cephalopod species pealei visual arrestins, occur in the same positions in the E. confirmed the presence of conserved amino acids, e.g., a lysine scolopes protein. in the seventh transmembrane region, presumably the site of retinal binding, a glycosylation site in the N terminus, a rho- Squid Retinal-Binding Protein. The EST database had a clone that encoded the C-terminal portion of the squid retinal-binding dopsin kinase phosphorylation site, and proline-rich repeats in protein (RALBP), with 82% identity in the region of alignment.
    [Show full text]
  • Chemoreception: a Consideration of Olfactory Signalling Proteins in Non-Olfactory Systems
    Chemoreception: A Consideration of Olfactory Signalling Proteins in Non-Olfactory Systems Yuliya Makeyeva A thesis in fulfillment of the requirements for the degree of Doctor of Philosophy School of Women’s and Children’s Health Faculty of Medicine November 2018 Thesis/Dissertation Sheet Australia's Global UNSWSYDNEY University Surname/Family Name Makeyeva Given Name/s Yuliya Abbreviation for degree as given in the University calendar PhD Faculty Faculty of Medicine School School of Women's and Children's Health Thesis Title Chemoreception: A Consideration of Olfactory Signalling Proteins in Non­ Olfactory Systems Abstract 350 words maximum: Chemoreception is a biological process whereby cells and/or organisms are stimulated by chemicals in their environment. The detectibn of chemicals triggers a response that can be the attraction or aversion by a single II cell or a highly integrative process that initiates a complex behaviour. Olfactory receptors (ORs) are a multigene family of molecules used to monitor extracellular chemical cues. They were originally described in the olfactory system as mediators of the sense of smell but have since been observed in a number of non-olfactory tissues. OR-mediated chemoreception in the olfactory system involves a G protein (Gait), adenylyl cyclase Ill (AC3), and OMP. ORs are difficultto study so little is known about their functions and expression profiles in these tissues. I, Olfactory marker protein (OMP) is a highly expressed protein found in mature olfactory sensory neurons of all vertebrates, co-labels with individual ORs, and is involved in intracellular signal transduction. OMP expression may indicate OR-mediated chemoreception in non-olfactory systems.
    [Show full text]
  • Documento Completo Descargar Archivo
    UNIVERSIDAD NACIONAL DE LA PLATA FACULTAD DE CIENCIAS EXACTAS DEPARTAMENTO DE CIENCIAS BIOLÓGICAS Trabajo de Tesis Doctoral: “Estudio del efecto del receptor de ghre- lina (GHSR) sobre la densidad en membrana de canales de calcio operados por voltaje (CaV) y sobre las propiedades biofísicas del sub- tipo CaV3”. Tesista: Lic. Emilio Román Mustafá Directora: Dra. Jesica Raingo Año: 2019 Agradecimientos Mi tesis doctoral fue uno de los mejores períodos de mi vida profesional y personal, y eso se lo debo a muchas de las personas que transitaron conmigo esta etapa. En primer lugar quiero agradecerle a Jesica, mi directora en esta tesis y mentora en mi carrera profesional. Gracias por permitirme ser parte de tu grupo de laboratorio cuando aún no tenía claro lo que significaba hacer un doctorado. Gracias por motivarme a hacer ciencia buena y de calidad, y sobretodo ciencia rigurosa y reproducible. Gracias por darnos a tus becarios y becarias la libertad de probar nuestras hipótesis cuando las pudimos ge- nerar. Gracias por despertar en nosotros el cuestionamiento a nuestros resultados y ajenos. Gracias por tu paciencia infinita cuando mil veces te he dicho “no entiendo”, gracias por exponerme a nuevos desafíos, y gracias por incentivarme a trabajar en equipo y entender lo valioso de hacer ciencia colectiva. Jesica es una investigadora a la que admiro, y sobre todo a la que le agradezco por ponderar de la misma manera hacer ciencia y formar buenos recursos humanos. También me gustaría agradecer a otras personas importantes en el laboratorio de Electrofisiología. Una de esas personas es Silvia, fue la primera que me enseñó lo que es trabajar en la mesada de un laboratorio realmente, teniendo mucha paciencia conmigo y con cualquier nuevo integrante.
    [Show full text]
  • Role of Molecular Chaperones in G Protein B5-Regulator of G Protein Signaling Dimer Assembly and G Protein by Dimer Specificity
    Brigham Young University BYU ScholarsArchive Theses and Dissertations 2009-04-02 Role of molecular chaperones in G protein B5-Regulator of G protein signaling dimer assembly and G protein By dimer specificity Alyson Cerny Howlett Brigham Young University - Provo Follow this and additional works at: https://scholarsarchive.byu.edu/etd Part of the Biochemistry Commons, and the Chemistry Commons BYU ScholarsArchive Citation Howlett, Alyson Cerny, "Role of molecular chaperones in G protein B5-Regulator of G protein signaling dimer assembly and G protein By dimer specificity" (2009). Theses and Dissertations. 2065. https://scholarsarchive.byu.edu/etd/2065 This Dissertation is brought to you for free and open access by BYU ScholarsArchive. It has been accepted for inclusion in Theses and Dissertations by an authorized administrator of BYU ScholarsArchive. For more information, please contact [email protected], [email protected]. ROLE OF MOLECULAR CHAPERONES IN G PROTEIN β5-REGULATOR OF G PROTEIN SIGNALING DIMER ASSEMBLY AND G PROTEIN βγ DIMER SPECIFICITY by Alyson Cerny Howlett A dissertation submitted to the faculty of Brigham Young University In partial fulfillment of the requirements for the degree of Doctor of Philosophy Department of Chemistry and Biochemistry Brigham Young University August 2009 BRIGHAM YOUNG UNIVERSITY GRADUATE COMMITTEE APPROVAL of a dissertation submitted by Alyson C. Howlett This dissertation has been read by each member of the following graduate committee and by majority vote has been found to be satisfactory. ____________________ __________________________________________ Date Barry M. Willardson, Chair ____________________ __________________________________________ Date Steven W. Graves ____________________ __________________________________________ Date Craig D. Thulin ____________________ __________________________________________ Date Allen R. Buskirk ____________________ __________________________________________ Date Laura C.
    [Show full text]
  • Regulation of Cardiac Hypertrophy and Metabolism by Regulator of G Protein Signalling 2 (RGS2)
    Western University Scholarship@Western Electronic Thesis and Dissertation Repository 9-1-2017 10:45 AM Regulation of Cardiac Hypertrophy and Metabolism by Regulator of G Protein Signalling 2 (RGS2) Katherine N. Lee The University of Western Ontario Supervisor Dr. Peter Chidiac The University of Western Ontario Co-Supervisor Dr. Qingping Feng The University of Western Ontario Graduate Program in Pharmacology and Toxicology A thesis submitted in partial fulfillment of the equirr ements for the degree in Doctor of Philosophy © Katherine N. Lee 2017 Follow this and additional works at: https://ir.lib.uwo.ca/etd Recommended Citation Lee, Katherine N., "Regulation of Cardiac Hypertrophy and Metabolism by Regulator of G Protein Signalling 2 (RGS2)" (2017). Electronic Thesis and Dissertation Repository. 4929. https://ir.lib.uwo.ca/etd/4929 This Dissertation/Thesis is brought to you for free and open access by Scholarship@Western. It has been accepted for inclusion in Electronic Thesis and Dissertation Repository by an authorized administrator of Scholarship@Western. For more information, please contact [email protected]. Abstract Pathological left ventricular hypertrophy is a maladaptive cardiomyocyte growth response to various cardiovascular conditions such as hypertension, and is a major risk factor for heart failure and stroke. The majority of drugs used to treat cardiovascular diseases target G protein coupled receptors (GPCRs), which are regulated by regulator of G protein signalling (RGS) proteins. RGS2 is a GTPase activating protein which limits Gq- and Gs-mediated signalling, which are known to play major roles in the development of pathological cardiac hypertrophy. In addition to its G protein effects, we have previously shown that RGS2 can also inhibit protein synthesis and cultured cardiomyocyte growth via a region in its RGS domain, RGS2eb, which inhibits the rate-limiting eIF2B/eIF2 step of protein synthesis initiation.
    [Show full text]
  • Download from DRYAD: 1246
    bioRxiv preprint doi: https://doi.org/10.1101/2021.01.10.426124; this version posted January 12, 2021. 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 4.0 International license. 1 Characterization of cephalic and non-cephalic sensory cell types 2 provides insight into joint photo- and mechanoreceptor 3 evolution 4 5 Roger Revilla-i-Domingo1,2,3, Vinoth Babu Veedin Rajan1,2, Monika 6 Waldherr1,2, Günther Prohaczka1,2, Hugo Musset1,2, Lukas Orel1,2, Elliot 7 Gerrard4, Moritz Smolka1,2,5, Matthias Farlik6,7, Robert J. Lucas4, Florian 8 Raible1,2,3,@ and Kristin Tessmar-Raible1,2,@ 9 10 1 Max Perutz Labs, University of Vienna, Vienna BioCenter, Dr. Bohr-Gasse 9/4, 1030 Vienna 11 2 Research Platform “Rhythms of Life”, University of Vienna, Vienna BioCenter, Dr. Bohr- 12 Gasse 9/4, A-1030 Vienna 13 3 Research Platform “Single-Cell Regulation of Stem Cells”, University of Vienna, 14 Althanstraße 14, A-1090 Vienna 15 4 Division of Neuroscience & Experimental Psychology, University of Manchester, UK 16 5 Center for Integrative Bioinformatics Vienna, Max Perutz Labs, University of Vienna and 17 Medical University of Vienna, Dr. Bohr-Gasse 9, Vienna, Austria 18 6 CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, 1090, 19 Vienna, Austria 20 7 Department of Dermatology, Medical University of Vienna, Währinger Gürtel 18-20, 1090 21 Vienna 22 23 24 @ Corresponding authors: 25 [email protected], 26 [email protected] 27 1 bioRxiv preprint doi: https://doi.org/10.1101/2021.01.10.426124; this version posted January 12, 2021.
    [Show full text]