DOCSLIB.ORG

And Insulin Receptor-Encoding Mrnas in Rainbow Trout

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
And Insulin Receptor-Encoding Mrnas in Rainbow Trout REGULATION OF INSULIN-AND INSULIN RECEPTOR-ENCODING mRNAS IN RAINBOW TROUT, Oncorhynchus mykiss A Dissertation Submitted to the Graduate Faculty of the North Dakota State University of Agriculture and Applied Science By Michael Alexander Caruso In Partial Fulfillment of the Requirements for the Degree of DOCTOR OF PHILOSOPHY Major Program: Cellular and Molecular Biology July 2011 Fargo, North Dakota North Dakota State University Graduate School Title REGULATION OF INSULIN-AND INSULIN RECEPTOR-ENCODING mRNAS IN RAINBOW TROUT, Oncorhynchus mykiss By Michael A. Caruso The Supervisory Committee certifies that this disquisition complies with North Dakota State University’s regulations and meets the accepted standards for the degree of DOCTOR OF PHILOSOPHY SUPERVISORY COMMITTEE: Dr. Mark Sheridan Chair Dr. Peggy Biga Dr. Wendy Reed Dr. Jane Schuh Approved: 7/29/2011 Dr. Wendy Reed Date Department Chair ABSTRACT In this work, rainbow trout were used as a model system to examine the regulation of insulin (INS)- and insulin receptor (IR)-encoding mRNA expression profiles. INS- and IR-encoding mRNAs were isolated, cloned, and sequenced; and shown to be differentially expressed within and among multiple tissue types. Regulation was examined through various nutritional and hormonal treatments (in vivo and in vitro). A real-time quantitative- PCR assay was developed to measure the respective levels of mRNA expression. Fasting, growth hormone (GH), and somatostatin (SS) differentially regulated INS and IR mRNAs within selected tissues, in vivo. Glucose, GH, SS, and insulin-like growth factor-1 (IGF-I) differentially regulated INS and IR mRNAs within selected tissues, in vitro. The results of this dissertation research display the identification and differential regulation of multiple INS- and IR-encoding mRNAs and suggest that independent mechanisms may serve to regulate the various isoforms in a tissue-specific manner. Future studies are also suggested. iii ACKNOWLEDGMENTS It is a pleasure to thank the many people without whom this dissertation work could not have been accomplished. First, I must give gratitude to my doctoral advisor, Dr. Mark Sheridan, who believed in me from day one and motivated me to pursue my career as a scientist. To say I look up to Mark is a gross understatement -- as a scholar and role model. Throughout my tenure at NDSU, Mark provided encouragement, sound advice, unparalleled teaching, and good company. I am deeply grateful to my graduate committee: Drs. Jane Schuh, Wendy Reed, and Peggy Biga for their elegant questions, feedback, and invaluable recommendations toward my dissertation research and future endeavors. This research was supported by several agencies. This project was funded by grants from the National Science Foundation, USA to M.A.S. North Dakota Experimental Program to Stimulate Competitive Research provided a Doctoral Dissertation Assistantship. The Department of Biological Sciences provided a teaching assistantship. I am forever indebted to the many research specialists and students who provided their support and camaraderie. Many of the technical achievements would not have been possible without the guidance of Jeff Kittilson. I am grateful for the technical assistance provided by Dr. Katie Reindl, Chad Walock, Rian Lee, Lincoln Martin, Felica Lamb, Sarah Danielson, Lindsey Norbeck, Elizabeth Ellens, Andrea Hanson, and Heather Bergan. I gratefully acknowledge Patrick Blaufus and Sujan Mamidi for their influence and expertise concerning phylogenetic analysis; and Curt Doetkott for statistical analysis. Also, I would like to thank Jeff and Chad for their insightful discussions on varied scientific and technical topics. iv Lastly, and most importantly, I would like to thank my family. My brother, Tony, who encouraged me to come to North Dakota and has never stopped pushing me to achieve my goals and beyond. My parents, Tom and Nancy, without their love and support, none of my dreams would ever be realized. And, to my son Brodyn, who I love and cherish every day. v TABLE OF CONTENTS ABSTRACT .......................................................................................................................... iii ACKNOWLEDGMENTS .................................................................................................... iv LIST OF TABLES .............................................................................................................. xiv LIST OF FIGURES ............................................................................................................. xv GENERAL INTRODUCTION .............................................................................................. 1 Insulin ................................................................................................................................. 3 Biosynthesis .................................................................................................................... 3 Structure .......................................................................................................................... 7 Evolution ...................................................................................................................... 11 Distribution ................................................................................................................... 14 Secretion ....................................................................................................................... 17 Function ........................................................................................................................ 20 Feeding/Fasting/Satiety ................................................................................................ 21 Growth .......................................................................................................................... 23 Development ................................................................................................................. 27 Intermediary Metabolism: Carbohydrates .................................................................... 28 Intermediary Metabolism: Lipids ................................................................................. 34 Nervous Tissue ............................................................................................................. 37 Insulin Receptor ............................................................................................................... 38 vi Biosynthesis .................................................................................................................. 39 Structure ........................................................................................................................ 40 Evolution ...................................................................................................................... 42 Distribution ................................................................................................................... 46 Binding ......................................................................................................................... 47 Signal Transduction ...................................................................................................... 49 References ........................................................................................................................ 57 OBJECTIVES ...................................................................................................................... 91 CHAPTER ONE. RAINBOW TROUT (Oncorhynchus mykiss) POSSESS TWO INSULIN-ENCODING mRNAS THAT ARE DIFFERENTIALLY EXPRESSED .......... 92 Abstract ............................................................................................................................ 92 Introduction ...................................................................................................................... 93 Materials and methods ..................................................................................................... 95 Experimental Animals .................................................................................................. 95 RNA Extraction ............................................................................................................ 96 Oligonucleotide Primers And Probes ........................................................................... 97 Isolation And Sequence Analysis Of Insulin-Encoding cDNAs .................................. 97 Preparation Of cDNA Standards .................................................................................. 98 Real-Time Reverse Transcription PCR ........................................................................ 99 Data Analysis .............................................................................................................. 100 vii Statistical Analyses ..................................................................................................... 100 Results ............................................................................................................................ 101 Characterization Of Two Insulin cDNAs ................................................................... 101 Differential Expression Of INS mRNAs Among And Within Tissues ...................... 102 Patterns Of INS mRNA Expression During Embryonic Development ...................... 103 Discussion .....................................................................................................................
Load more

Recommended publications
  • Chemistry of Pancreatic Polypeptide Hormone with Official Preparation
    wjpmr, 2020,6(9), 102-114 SJIF Impact Factor: 5.922 WORLD JOURNAL OF PHARMACEUTICAL Review Article Arpan et al. World Journal of Pharmaceutical and Medical Research AND MEDICAL RESEARCH ISSN 2455-3301 www.wjpmr.com Wjpmr CHEMISTRY OF PANCREATIC POLYPEPTIDE HORMONE WITH OFFICIAL PREPARATION Arpan Chanda*1, Arunava Chandra Chandra2, Dr. Dhrubo Jyoti Sen2 and Dr. Dhananjoy Saha3 1Department of Pharmaceutical Chemistry, Netaji Subhas Chandra Bose Institute of Pharmacy, Roypara, Chakdaha Dist–Nadia, Pin‒741222, West Bengal, India. 2Department of Pharmaceutical Chemistry, School of Pharmacy, Techno India University, Salt‒Lake City, Sector‒V, EM‒4, Kolkata‒700091, West Bengal, India. 3Deputy Director of Technical Education, Directorate of Technical Education, Bikash Bhavan, Salt Lake City, Kolkata‒700091, West Bengal, India. *Corresponding Author: Arpan Chanda Department of Pharmaceutical Chemistry, Netaji Subhas Chandra Bose Institute of Pharmacy, Roypara, Chakdaha Dist–Nadia, Pin‒741222, West Bengal, India. Article Received on 05/07/2020 Article Revised on 26/07/2020 Article Accepted on 16/08/2020 ABSTRACT Insulin which is a peptide hormone is produced by β‒cells of the pancreatic islets and it is considered to be the main anabolic hormone of the body. It further regulates the metabolism of carbohydrates, fats and protein by promoting the absorption of glucose from the blood into liver, fat and skeletal muscle cells. In these tissues the absorbed glucose from the blood is thus converted into either glycogen via glycogenesis or fats (triglycerides) via lipogenesis, or, in the case of the liver both glycogenesis and lipogenesis. Glucose production and secretion by the liver is strongly supported by high concentrations of insulin in the blood.
    [Show full text]
  • Identification, Characterization and Application of Autoantigens in Type 1 Diabetes Mellitus
    IDENTIFICATION, CHARACTERIZATION AND APPLICATION OF AUTOANTIGENS IN TYPE 1 DIABETES MELLITUS Henk-Jan Aanstoot IDENTIFICATION, CHARACTERIZATION AND APPLICATION OF AUTOANTIGENS IN TYPE 1 DIABETES MELLITUS IDENTIFICATIE, KARAKTERISERING EN TOEPASSINGEN VAN AUTOANTIGENEN BIJ TYPE 1 DIABETES MELLITUS PROEFSCHRIFT TER VERKRIJGING VAN DE GRAAD VAN DOCTOR AAN DE ERASMUS UNIVERSITEIT ROTTERDAM OP GEZAG VAN DE RECTOR MAGNIFICUS Prof. Dr. P.WC. AKKERMANS M.Lit EN VOLGENS BESLUIT VAN HET COLLEGE VAN DEKANEN. DE OPENBARE VERDEDIGING ZAL PLAATSVINDEN OP VRIJDAG 5 NOVEMBER 1993 OM 16.00 UUR DOOR HENDRIK-JAN AANSTOOT GEBOREN TE DEVENTER OFFSETDRUKKEA!J HAVEKA BV, ALBL.ASSERDAM PROMOTIECOMMISSIE PROMOTORES: Prof. Dr. H.K.A. Visser Prof. Dr. H. Galjaard CO-PROMOTOR: Dr. G.J. Bruining OVERIGE LEDEN: Prof. Dr. S.W.J. Lamberts Prof Dr. R. Benner The work presented in this thesis was generously supported by grants from the "Ter Meulen Fonds" (Koninklijke Nederlandse Academie van Wetenschappen), the "Albert Renold Fellowship" of the European Association for the Study of Diabetes (EASD), the Sophia Foundation for Medical Research (SSWO), the Child Health and Wellbeing Fund, Rotterdam and Novo-Nordisk A/S, Denmark. Additional generous support was given by the Rotterdam Council of Clinical Genetics (Stichting Klinische Genetica Rotterdam). Printing of this thesis was possible with generous support form Novo-Nordisk, Nederland B. V. 'Door de vreugde van de ontdekking verblind, zullen wij niet uit het oog verliezen, dat het insuline nog s/echts een eerste schrede is op den weg eener oorzakelijke behandeling der suikerziekte' A.A. Hijmans van den Berg, 1925, Utrecht , Voordrachten over suikerziekte, 278. opgedragen aan de grate mensen van morgen 7 CONTENTS OUTLINE AND AIMS OF THIS THESIS 11 Chapter 1 GENERAL INTRODUCTION 13 1 .1 Diabetes Mellitus.
    [Show full text]
  • Anatomic and Molecular Characterization of the Endocrine
    Fortin et al. Zoological Studies (2015) 54:21 DOI 10.1186/s40555-014-0093-4 RESEARCH Open Access Anatomic and molecular characterization of the endocrine pancreas of a teleostean fish: Atlantic wolffish (Anarhichas lupus) Jessica S Fortin1*, Ariane Santamaria-Bouvier2, Stéphane Lair2, André D Dallaire1 and Marie-Odile Benoit-Biancamano1* Abstract Background: The biologic attributes of the endocrine pancreas and the comparative endocrinology of islet amyloid polypeptide (IAPP) of fish are not well described in the literature. This study describes the endocrine pancreas of one teleostean fish. Ten captive Atlantic wolffish (Anarhichas lupus) from the Montreal Biodome were submitted for necropsy and their pancreata were collected. Results: Grossly, all the fish pancreata examined contained 1-3 nodules of variable diameter (1-8 mm). Microscopically, the nodules were uniform, highly cellular, and composed of polygonal to elongated cells. Immunofluorescence for pancreatic hormones was performed. The nodules were immunoreactive for insulin most prominent centrally, but with IAPP and glucagon only in the periphery of the nodules. Exocrine pancreas was positive for chromogranin A. Not previously recognized in fish, IAPP immunoreactivity occurred in α, glucagon-containing, cells and did not co-localize with insulin in β cells. The islet tissues were devoid of amyloid deposits. IAPP DNA sequencing was performed to compare the sequence among teleost fish and the potency to form amyloid fibrils. In silico analysis of the amino acid sequences 19-34 revealed that it was not amyloidogenic. Conclusions: Amyloidosis of pancreatic islets would not be expected as a spontaneous disease in the Atlantic wolffish. Our study underlines that this teleost fish is a potential candidate for pancreatic xenograft research.
    [Show full text]
  • Ontogenetic and Phylogenetic Development of the Endocrine Pancreas (Islet Organ) in Fishes
    General and Comparative Endocrinology 116, 303–335 (1999) Article ID gcen.1999.7376, available online at http://www.idealibrary.com on REVIEW Ontogenetic and Phylogenetic Development of the Endocrine Pancreas (Islet Organ) in Fishes John H. Youson and Azza A. Al-Mahrouki Department of Zoology and Division of Life Sciences, University of Toronto at Scarborough, Scarborough, Ontario M1C 1A4 Canada Accepted August 25, 1999 The morphology of the gastroenteropancreatic (GEP) relatively small islets in the generalized euteleosts and system of fishes was reviewed with the objective of the tendency for the concentration into Brockmann providing the phylogenetic and ontogenetic development bodies of large (principal) islets (with or without second- of the system in this vertebrate group, which includes ary islets) in the more derived forms. The holostean agnathans and gnathostome cartilaginous, actinoptyeryg- actinopterygians (Amiiformes and Semiontiformes) share ian, and sarcopterygian fishes. Particular emphasis is with the basal teleosts (osteoglossomorphs, elopo- placed on the fish homolog of the endocrine pancreas of morphs) the diffuse arrangement of the components of other vertebrates, which is referred to as the islet organ. the islet organ that is seen in generalized euteleosts. The one-hormone islet organ (B cells) of larval lampreys Since principal islets are also present in adult lampreys is the most basic pattern seen among a free-living the question arises whether principal islets are a derived vertebrate, with the two-hormone islet organ (B and D or a generalized feature among teleosts. There is a cells) of hagfish and the three-hormone islet organ (B, D, paucity of studies on the ontogeny of the GEP system in and F cells) of adult lampreys implying a phylogenetic fish but it has been noted that the timing of the trend toward the classic four-hormone islet tissue (B, D, appearance of the islet cell types parallels the time that F,and A cells) in most other fishes.
    [Show full text]
  • Xenotransplantation of Caprine Pancreatic Islets in Diabetic Mice
    UNIVERSITI PUTRA MALAYSIA XENOTRANSPLANTATION OF CAPRINE PANCREATIC ISLETS IN DIABETIC MICE HOMAYOUN HANI IB 2013 27 XENOTRANSPLANTATION OF CAPRINE PANCREATIC ISLETS IN DIABETIC MICE UPM By HOMAYOUN HANI ThesisCOPYRIGHT Submitted to the School of Graduate Studies, Universiti Putra Malaysia, in Fulfillment of the Requirements for the Degree of Doctor of Philosophy © December 2013 i DEDICATION I dedicate with appreciation to my parents, Javad and Maryam to my brothers, Reza and Hadi to my beloved wife, Kazhal Sarsaifi, for their great support, patience, and motivation UPM COPYRIGHT © ii Abstract of thesis presented to the Senate of Universiti Putra Malaysia in fulfillment of the requirement for the degree of Doctor of Philosophy XENOTRANSPLANTATION OF CAPRINE PANCREATIC ISLETS IN DIABETIC MICE By HOMAYOUN HANI December 2013 Chairman : Zeenathul Nazariah Allaudin, PhD Faculty : Institute of Bioscience Diabetes is one of the aggressive metabolic syndromes, which allied with high levels of blood glucose subsequent of imperfections in insulin production thatUPM causes glucose to increase in the body. For decades, excellent improvement has been obtained in clinical transplantation of pancreatic islet. Because of that, islet transplantation has become the main alternative cure for type 1 diabetics. However, important obstacles still remain to use of islet transplants routinely as a treatment choice. The deficiency of human islet donors makes the search for alternative islet sources mandatory for future developments in pancreatic islet transplantation. Venturing into xenotransplantation provides tremendous extensive sources of islet cells. The improvement of genetically engineered pigs expressing human complement regulatory proteins to defeat immune damaging pathways has been promising; however, the necessity of massive immunosuppressive and the concern for zoonotic viral transmission are some of the constraints.
    [Show full text]
  • Characterization of the Gastro-Entero-Pancreatic System of Osteoglossomorpha: an Immunohistochemical, Immunocytochemical, and Molecular Study
    Characterization of the gastro-entero-pancreatic system of Osteoglossomorpha: an immunohistochemical, immunocytochemical, and molecular study Azza Abdul Fattah Al-Mahrouki A thesis submitted in conformity with the requirements for the degree of Doctor of Philosophy in the Department of Zoology University of toronto @ a Al-Mahrouki, 2001 National Library Bibliothèque nationale 1*1 of Canada du Canada Acquisitions and Acquisitions et Bibliographie Services services bibliographiques 395 Wellington Street 395. nie Wellington Ottawa ON KI A ON4 Ottawa ON KIA ON4 Canada Canada vow& Yom réienww Our fi& Notre réftirence The author has granted a non- L'auteur a accordé une licence non exclusive licence allowing the exclusive permettant à la National Lïbrary of Canada to Bibliothèque nationale du Canada de reproduce, loan, distri'bute or sell reproduire, prêter, distribuer ou copies of this thesis in microform, vendre des copies de cette thèse sous paper or electronic formats. la forme de microfiche/nlm, de reproduction sur papier ou sur format électronique. The author retains ownership of the L'auteur conserve la propriété du copyright in thesis. Neither the droit d'auteur qui protège cette thèse. thesis nor substantial extracts fiom it Ni la thèse ni des extraits substantiels may be printed or otherwise de celle-ci ne doivent être imprimés reproduced without the author's ou autrement reproduits sans son permission. autorisation- Abstract Characterization of the gastro-entero-pancreatic system of Osteoglossomorpha: An immunohistochemical, immunocytochemical and molecular study. Doctor of Philosophy, 2001- Azza Abdul Fattah Al-Mahrouki. Department of Zoology. University of Toronto. The principal objectives of this study were twofold: fjrst, to compare the distrr'bution and structure of the GEP system of the osteoglossomorphs with that of the other teleosts and with vertebrates in general; second, to examine the possibility of using the primary structure of the GEP system peptides in a phylogenetic analysis.
    [Show full text]
  • What Hormones May Regulate Food Intake in Fish?
    Ayuut. Living Resour.. 1997, 10, 37 1-379 What hormones may regulate food intake in fish? Pierre-Yves Le Bail (') and Gilles Rteiif2) (') 1,nhoratoir~d~ Phytio1og-i~rkr Poissons, INRA, Campus de Umulieu, 35042 I~L~LP~,Fruncp. 1;-mail: [email protected]./r '2' Laboratoire de PhysiolobIz'e des Poissons, ibI~M;IcK. I'. 70, 29280 l'louz~n6,France. E.:-mail: ghoeu/@ijiremer.j? Received May 2, 1997; acccptcd October 23, 1997. Le Bail P.-Y., G. Bœuf. Aquat. Livin~Resour., 1997, 10,371-379. Abstract This is an ovcmicw of thc hormoncs which may be involved in food intake control in fish, and some hypo- thetical pathways of thcir action arc givcn based on mammalian knowledge. Most of the observed effects of these hormones may result from four types of mechanisms, each hormone acting by one or several as follows: (1) hormones could have a direct effect on central nervous system centres, associated with food intake behaviour or via vagal afferent neurons; (2) an indirect effect may occur via the gut which slows gastrointestinal transit, thus rcsulting in stomach distention which activates vagal afferent neurons; (3) they could have an indirect cffcct, acting dircctly on intermediary metabolism via glucose, free fatty acids or amino acids mohilization or storage; (4) the last possible pathway is an indirect effect by modifying directly or indirectly secretions of other hormones involved in food intake control. Some of thesc hormones (CCK, PYY, glucagon, adrenalin) act as short-term factors which regulate meal ingestion and are generally inhibi- tory factors.
    [Show full text]