DOCSLIB.ORG

Calcium Regulation of Cell-Cell Communication and Extracellular Signaling

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Calcium Regulation of Cell-Cell Communication and Extracellular Signaling Georgia State University ScholarWorks @ Georgia State University Chemistry Dissertations Department of Chemistry 8-12-2016 CALCIUM REGULATION OF CELL-CELL COMMUNICATION AND EXTRACELLULAR SIGNALING Juan Zou Follow this and additional works at: https://scholarworks.gsu.edu/chemistry_diss Recommended Citation Zou, Juan, "CALCIUM REGULATION OF CELL-CELL COMMUNICATION AND EXTRACELLULAR SIGNALING." Dissertation, Georgia State University, 2016. https://scholarworks.gsu.edu/chemistry_diss/123 This Dissertation is brought to you for free and open access by the Department of Chemistry at ScholarWorks @ Georgia State University. It has been accepted for inclusion in Chemistry Dissertations by an authorized administrator of ScholarWorks @ Georgia State University. For more information, please contact [email protected]. CALCIUM REGULATION OF CELL-CELL COMMUNICATION AND EXTRACELLULAR SIGNALING by pro JUAN ZOU Under the Direction of Jenny Yang, PhD ABSTRACT As a highly versatile signal, Ca2+ operates over a wide temporal range to regulate many different cellular processes, impacting nearly every aspect of cellular life including excitability, exocytosis, motility, apoptosis, and transcription. While it has been well recognized that Ca2+ acts as both a second messenger to regulate cell-cell communication upon external stimuli and as a first messenger to integrate extracellular with intracellular signaling in various cell types. Molecular bases for such regulation and related human diseases are largely hampered by the challenges related to key membrane proteins. In the present study, we first investigated the regulatory role of 2+ 2+ 2+ intracellular Ca ([Ca ]i) on Connexin45 (Cx45) gap junction through a ubiquitous Ca sensor protein-Calmodulin (CaM). Using bioluminescence resonance energy transfer assay, this study provides the first evidence of direct association of Cx45 and CaM in a Ca2+-dependent manner in cells. Complementary approaches including bioinformatics analysis and various biophysical methods identified a putative CaM-binding site in the intracellular loop of Cx45 with high Ca2+/CaM-binding affinity and Ca2+-dependent binding mode that is different from alpha family of connexins. To understand the role of extracellular calcium in regulation of gap junction hemichannels, we would like to prove a possible Ca2+-binding site predicted by our computational algorithm MUGSR in Connexin 26 (Cx26) through mutagenesis study, metal binding affinity measurement, conformational changes examination of purified Cx26 protein from Sf9; however, we failed to achieve this goal due to either the limitation of available methods or lethal effect of mutating the predicted Ca2+-binding ligand. Additionally, in this study, we identified a putative Ca2+-binding site in metabotropic glutamate receptor 5 (mGluR5) and demonstrated the importance of this Ca2+-binding site in activation of mGluR5 and modulating the actions of other orthosteric ligands on mGluR5. In addition, we successfully solved the first crystal structure of the extracellular domain of Ca2+-sensing receptor (CaSR) bound with Mg2+ and an unexpected Trp derivative. The extensive study of mechanism of CaSR function specifically through Mg2+-binding site and the unexpected ligand-binding site was done using several cell-based assays in wild type CaSR and mutants. Studies in this dissertation provides more information on how Ca2+ regulates gap junction channels, modulates mGluR5 activities and structural basis for regulation of CaSR by Mg2+ and an unexpected Trp derivative co-agonist. INDEX WORDS: Ca2+; Mg2+; CaM; Cx45; gap junction; mGluR5; CaSR. CALCIUM REGULATION OF CELL-CELL COMMUNICATION AND EXTRACELLULAR SIGNALING by JUAN ZOU A Dissertation Submitted in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy in the College of Arts and Sciences Georgia State University 2016 Copyright by Juan Zou 2016 CALCIUM REGULATION OF CELL-CELL COMMUNICATION AND EXTRACELLULAR SIGNALING by JUAN ZOU Committee Chair: Jenny Yang Committee: Jun Yin Ivaylo Ivanov Electronic Version Approved: Office of Graduate Studies College of Arts and Sciences Georgia State University May 2016 iv DEDICATION I want first of all to thank our previous lab members who paved the path before me and provided support for me to stand to extend the work. Without the method they established in our lab, I could not achieve all these. This is also a tribute to my loving family and friends who supported me on this journey. Their thoughtfulness and kindness always warms me. Their constant encouragement always ring in my ears. They have never left my side no matter how many mistakes I have made. Their generous tolerance promote me to be grateful. I would like to extend my gratitude to the Brain and Behavior fellowship from Georgia State Univeristy. Without it, I could not finish my PhD study here. The great seminar classes provided by Brain and Behavior also introduced me to the mysterious and interesting neuron science field. v ACKNOWLEDGEMENTS First and foremost, I would like to express my sincere gratitude to my advisor Professor Dr. Jenny Yang for the continuous support of my Ph. D study and research. I thank her for introducing me to this wonderful research world. Her immense knowledge, patience guidance, continuous encouragement allows me to grow as a research scientist. Her enthusiasm, strong thirst for knowledge and motivation always inspire me. She has written countless proposals in the past six years in order to obtain continuous support from NIH and to survive in the competitive academic field. I am motivated by the countless sleepless nights she spent on reading and writing papers, numerous times she sat together with us to discuss the experimental plans, joined us to learn from outside experts and she proudly introduced her students to others in conferences. She has set a good example for me. I could not have imagined having a better advisor and mentor for my Ph. D study. Not only in research, but she also has given me lots of unselfish help, motherly love and forgiveness in life. She is always a genuinely caring and helpful person. She is always the first one that I can share my feelings with, the first shoulder I can cry on, and the first one who gets me through my troubles. Her advice on both my research as well as on my life have been invaluable. Besides my advisor, I would like to thank the rest of my committee members, Dr. Jun Yin and Dr. Ivaylo Ivanov for serving as my committee members. Their insightful comments, brilliant suggestions, and encouragement helped me all the time in my research and writing of this dissertation. Thank you for making my defense an enjoyable experience. The hard questions in the defense sparked new ideas and invigorated me to widen my research. vi My sincere gratitude also goes to Drs. Richard Veenstra and Edward M. Brown. Their profound knowledge in gap junction and G-protein coupled receptor field is always like the lighthouse, which gives me hope and guidance when I am floating like the boat on the ocean of knowledge. During my Ph. D study, especially in the first few years, they are my encyclopedia. Discussions with them always help me solve problems and puzzles, and enlightened me. I will never forget their support and inspiration. I am particularly indebted to our collaborators: Dr. John Helper from Emory, Dr. Jian Hu from Michigan State University, and Dr. Kelly Moremen from University of Georgia. In 2013, my research was at standstill and I could not prove the interaction between calmodulin and Cx45 in the living cell. We received am email from Dr. Helper asking collaboration on their RGS14 project. In the presentation they gave in our lab, I was so excited to realize that their BRET assay can be used in my experiment to monitor protein-protein interaction in vivo. Nicole from Dr. Helper’s lab sent me the related plasmid and gave me detailed training on performing the BRET assay. They also kindly allowed to culture cells in their lab and use their microplate reader. With their help, we finally could obtained promising result. Without their precious support it would not be possible to conduct the Cx45 project. Furthermore, both Dr. Hu’s and Dr. Moremen’s group contributed extensively to the CaSR project. Their expertise in membrane protein expression in mammalian cells and crystallization moved our CaSR project a significant step forward. Without them, we would not be successful to solve the CaSR-ECD structure and identified an unexpected CaSR ligand. Words cannot express how grateful I am to all of them. I am grateful to Drs. Yanyi Chen and Chen Zhang for their patient hands-on teaching and training. Dr. Chen introduced me to the gap junction field when I first joined in Dr. Yang’s group vii in 2010. I learnt all important biophysical technique from him, applying what I have learned from him into research. His critical thinking and quick learning also impressed me a lot. I am grateful to Dr. Chen for enlightening me with the first glance of research. In my second year of my Ph. D study, I was attracted by the wide application of cell studies in research. Therefore, I turned to Dr. Zhang and asked her to train me. She said “yes” to me without any hesitation and became my second mentor in Dr. Yang’s lab. How lucky I am to have them as my mentors. Their training and advice on both research as well as on my career are priceless. I would also like to thank all of the rest labmates and friends, particularly Jie Jiang, Jinjuan Qiao, Yusheng Jiang, You Zhuo, Jie Feng, Li Zhang, Mani Salarian, Rakshya Gokhali and Cassie Miller, for the stimulating discussions, for the time we were working together to meet deadlines, and for the fun we had together in the past six years. They are also my close friends in my life, who were always aside to support me.
Load more

Recommended publications
  • Viewed Under 23 (B) Or 203 (C) fi M M Male Cko Mice, and Largely Unaffected Magni Cation; Scale Bars, 500 M (B) and 50 M (C)
    BRIEF COMMUNICATION www.jasn.org Renal Fanconi Syndrome and Hypophosphatemic Rickets in the Absence of Xenotropic and Polytropic Retroviral Receptor in the Nephron Camille Ansermet,* Matthias B. Moor,* Gabriel Centeno,* Muriel Auberson,* † † ‡ Dorothy Zhang Hu, Roland Baron, Svetlana Nikolaeva,* Barbara Haenzi,* | Natalya Katanaeva,* Ivan Gautschi,* Vladimir Katanaev,*§ Samuel Rotman, Robert Koesters,¶ †† Laurent Schild,* Sylvain Pradervand,** Olivier Bonny,* and Dmitri Firsov* BRIEF COMMUNICATION *Department of Pharmacology and Toxicology and **Genomic Technologies Facility, University of Lausanne, Lausanne, Switzerland; †Department of Oral Medicine, Infection, and Immunity, Harvard School of Dental Medicine, Boston, Massachusetts; ‡Institute of Evolutionary Physiology and Biochemistry, St. Petersburg, Russia; §School of Biomedicine, Far Eastern Federal University, Vladivostok, Russia; |Services of Pathology and ††Nephrology, Department of Medicine, University Hospital of Lausanne, Lausanne, Switzerland; and ¶Université Pierre et Marie Curie, Paris, France ABSTRACT Tight control of extracellular and intracellular inorganic phosphate (Pi) levels is crit- leaves.4 Most recently, Legati et al. have ical to most biochemical and physiologic processes. Urinary Pi is freely filtered at the shown an association between genetic kidney glomerulus and is reabsorbed in the renal tubule by the action of the apical polymorphisms in Xpr1 and primary fa- sodium-dependent phosphate transporters, NaPi-IIa/NaPi-IIc/Pit2. However, the milial brain calcification disorder.5 How- molecular identity of the protein(s) participating in the basolateral Pi efflux remains ever, the role of XPR1 in the maintenance unknown. Evidence has suggested that xenotropic and polytropic retroviral recep- of Pi homeostasis remains unknown. Here, tor 1 (XPR1) might be involved in this process. Here, we show that conditional in- we addressed this issue in mice deficient for activation of Xpr1 in the renal tubule in mice resulted in impaired renal Pi Xpr1 in the nephron.
    [Show full text]
  • Metabotropic Glutamate Receptors
    mGluR Metabotropic glutamate receptors mGluR (metabotropic glutamate receptor) is a type of glutamate receptor that are active through an indirect metabotropic process. They are members of thegroup C family of G-protein-coupled receptors, or GPCRs. Like all glutamate receptors, mGluRs bind with glutamate, an amino acid that functions as an excitatoryneurotransmitter. The mGluRs perform a variety of functions in the central and peripheral nervous systems: mGluRs are involved in learning, memory, anxiety, and the perception of pain. mGluRs are found in pre- and postsynaptic neurons in synapses of the hippocampus, cerebellum, and the cerebral cortex, as well as other parts of the brain and in peripheral tissues. Eight different types of mGluRs, labeled mGluR1 to mGluR8, are divided into groups I, II, and III. Receptor types are grouped based on receptor structure and physiological activity. www.MedChemExpress.com 1 mGluR Agonists, Antagonists, Inhibitors, Modulators & Activators (-)-Camphoric acid (1R,2S)-VU0155041 Cat. No.: HY-122808 Cat. No.: HY-14417A (-)-Camphoric acid is the less active enantiomer (1R,2S)-VU0155041, Cis regioisomer of VU0155041, is of Camphoric acid. Camphoric acid stimulates a partial mGluR4 agonist with an EC50 of 2.35 osteoblast differentiation and induces μM. glutamate receptor expression. Camphoric acid also significantly induced the activation of NF-κB and AP-1. Purity: ≥98.0% Purity: ≥98.0% Clinical Data: No Development Reported Clinical Data: No Development Reported Size: 10 mM × 1 mL, 100 mg Size: 10 mM × 1 mL, 5 mg, 10 mg, 25 mg (2R,4R)-APDC (R)-ADX-47273 Cat. No.: HY-102091 Cat. No.: HY-13058B (2R,4R)-APDC is a selective group II metabotropic (R)-ADX-47273 is a potent mGluR5 positive glutamate receptors (mGluRs) agonist.
    [Show full text]
  • A Computational Approach for Defining a Signature of Β-Cell Golgi Stress in Diabetes Mellitus
    Page 1 of 781 Diabetes A Computational Approach for Defining a Signature of β-Cell Golgi Stress in Diabetes Mellitus Robert N. Bone1,6,7, Olufunmilola Oyebamiji2, Sayali Talware2, Sharmila Selvaraj2, Preethi Krishnan3,6, Farooq Syed1,6,7, Huanmei Wu2, Carmella Evans-Molina 1,3,4,5,6,7,8* Departments of 1Pediatrics, 3Medicine, 4Anatomy, Cell Biology & Physiology, 5Biochemistry & Molecular Biology, the 6Center for Diabetes & Metabolic Diseases, and the 7Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202; 2Department of BioHealth Informatics, Indiana University-Purdue University Indianapolis, Indianapolis, IN, 46202; 8Roudebush VA Medical Center, Indianapolis, IN 46202. *Corresponding Author(s): Carmella Evans-Molina, MD, PhD ([email protected]) Indiana University School of Medicine, 635 Barnhill Drive, MS 2031A, Indianapolis, IN 46202, Telephone: (317) 274-4145, Fax (317) 274-4107 Running Title: Golgi Stress Response in Diabetes Word Count: 4358 Number of Figures: 6 Keywords: Golgi apparatus stress, Islets, β cell, Type 1 diabetes, Type 2 diabetes 1 Diabetes Publish Ahead of Print, published online August 20, 2020 Diabetes Page 2 of 781 ABSTRACT The Golgi apparatus (GA) is an important site of insulin processing and granule maturation, but whether GA organelle dysfunction and GA stress are present in the diabetic β-cell has not been tested. We utilized an informatics-based approach to develop a transcriptional signature of β-cell GA stress using existing RNA sequencing and microarray datasets generated using human islets from donors with diabetes and islets where type 1(T1D) and type 2 diabetes (T2D) had been modeled ex vivo. To narrow our results to GA-specific genes, we applied a filter set of 1,030 genes accepted as GA associated.
    [Show full text]
  • Src Regulation of Cx43 Phosphorylation and Gap Junction Turnover
    biomolecules Article Src Regulation of Cx43 Phosphorylation and Gap Junction Turnover Joell L. Solan 1 and Paul D. Lampe 1,2,* 1 Translational Research Program, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA; [email protected] 2 Department of Global Health, Pathobiology Program, University of Washington, Seattle, WA 98109, USA * Correspondence: [email protected] Received: 27 October 2020; Accepted: 22 November 2020; Published: 24 November 2020 Abstract: The gap junction protein Connexin43 (Cx43) is highly regulated by phosphorylation at over a dozen sites by probably at least as many kinases. This Cx43 “kinome” plays an important role in gap junction assembly and turnover. We sought to gain a better understanding of the interrelationship of these phosphorylation events particularly related to src activation and Cx43 turnover. Using state-of-the-art live imaging methods, specific inhibitors and many phosphorylation-status specific antibodies, we found phospho-specific domains in gap junction plaques and show evidence that multiple pathways of disassembly exist and can be regulated at the cellular and subcellular level. We found Src activation promotes formation of connexisomes (internalized gap junctions) in a process involving ERK-mediated phosphorylation of S279/282. Proteasome inhibition dramatically and rapidly restored gap junctions in the presence of Src and led to dramatic changes in the Cx43 phospho-profile including to increased Y247, Y265, S279/282, S365, and S373 phosphorylation. Lysosomal inhibition, on the other hand, nearly eliminated phosphorylation on Y247 and Y265 and reduced S368 and S373 while increasing S279/282 phosphorylation levels. We present a model of gap junction disassembly where multiple modes of disassembly are regulated by phosphorylation and can have differential effects on cellular signaling.
    [Show full text]
  • Genes in Eyecare Geneseyedoc 3 W.M
    Genes in Eyecare geneseyedoc 3 W.M. Lyle and T.D. Williams 15 Mar 04 This information has been gathered from several sources; however, the principal source is V. A. McKusick’s Mendelian Inheritance in Man on CD-ROM. Baltimore, Johns Hopkins University Press, 1998. Other sources include McKusick’s, Mendelian Inheritance in Man. Catalogs of Human Genes and Genetic Disorders. Baltimore. Johns Hopkins University Press 1998 (12th edition). http://www.ncbi.nlm.nih.gov/Omim See also S.P.Daiger, L.S. Sullivan, and B.J.F. Rossiter Ret Net http://www.sph.uth.tmc.edu/Retnet disease.htm/. Also E.I. Traboulsi’s, Genetic Diseases of the Eye, New York, Oxford University Press, 1998. And Genetics in Primary Eyecare and Clinical Medicine by M.R. Seashore and R.S.Wappner, Appleton and Lange 1996. M. Ridley’s book Genome published in 2000 by Perennial provides additional information. Ridley estimates that we have 60,000 to 80,000 genes. See also R.M. Henig’s book The Monk in the Garden: The Lost and Found Genius of Gregor Mendel, published by Houghton Mifflin in 2001 which tells about the Father of Genetics. The 3rd edition of F. H. Roy’s book Ocular Syndromes and Systemic Diseases published by Lippincott Williams & Wilkins in 2002 facilitates differential diagnosis. Additional information is provided in D. Pavan-Langston’s Manual of Ocular Diagnosis and Therapy (5th edition) published by Lippincott Williams & Wilkins in 2002. M.A. Foote wrote Basic Human Genetics for Medical Writers in the AMWA Journal 2002;17:7-17. A compilation such as this might suggest that one gene = one disease.
    [Show full text]
  • Hearing Loss Carrie Crain, B.S
    R.C.P.U. NEWSLETTER Editor: Heather J. Stalker, M.Sc. Director: Roberto T. Zori, M.D. R.C. Philips Research and Education Unit Vol. XIX No. 2 A statewide commitment to the problems of mental retardation January 2008 R.C. Philips Unit ♦ Division of Pediatric Genetics, Box 100296 ♦ Gainesville, FL 32610 ♦ (352)392-4104 E Mail: [email protected]; [email protected] Website: http://www.peds.ufl.edu/divisions/genetics/newsletters.htm Hearing Loss Carrie Crain, B.S. Regional Follow-Up Coordinator, Newborn Hearing Screening Introduction Phase 1: Screening It is estimated that 70 million people world-wide, or 1% of the All newborns are screened for hearing loss shortly after birth world’s population have some form of hearing loss that affects using either evoked otoacoustic emissions (EOAE) or auditory language communication. Profound congenital hearing loss is brainstem response testing (ABR) to look for permanent estimated to affect 1 in 1000 births (Tekin et al. 2001). If bilateral, unilateral, sensory or conductive hearing loss significant hearing loss is not detected early in life, it can have averaging 30-40 dB or more. negative impacts on speech, language and cognitive ABR testing uses a series of clicks to evoke responses, development. originating in the eighth cranial nerve and auditory brainstem and recorded by electrodes placed on the head. EOAE testing Newborn Screening uses a probe inserted into the ear to measure sounds The history of newborn screening dates back to the 1960’s originating within the cochlea. EOAEs reflect the activity of the when newborns were first screened for phenylketonuria (PKU).
    [Show full text]
  • Type of PKD1 Mutation Influences Renal Outcome in ADPKD
    CLINICAL RESEARCH www.jasn.org Type of PKD1 Mutation Influences Renal Outcome in ADPKD † †‡ †‡ Emilie Cornec-Le Gall,* Marie-Pierre Audrézet, Jian-Min Chen, Maryvonne Hourmant,§ | †† Marie-Pascale Morin, Régine Perrichot,¶ Christophe Charasse,** Bassem Whebe, ‡‡ || † Eric Renaudineau, Philippe Jousset,§§ Marie-Paule Guillodo, Anne Grall-Jezequel,* †‡ †‡ † Philippe Saliou, Claude Férec, and Yannick Le Meur* *Nephrology Department, University Hospital, Brest, France; †Université de Bretagne Occidentale and European University of Brittany, Brest, France; ‡Molecular Genetic Department, INSERM U1078, University Hospital, EFS- Bretagne, Brest, France; §Nephrology Department, University Hospital, Nantes, France; |Nephrology Department, University Hospital, Rennes, France; ¶Nephrology Department, Vannes Hospital, Vannes, France; **Nephrology Department, Saint Brieuc Hospital, Saint Brieuc, France; ††Nephrology Department, Quimper Hospital, Quimper, France; ‡‡Nephrology Department, Broussais Hospital, Saint Malo, France; §§Nephrology Department, Pontivy Hospital, Pontivy, France; and ||Association des Urémiques de Bretagne, Brest, France ABSTRACT Autosomal dominant polycystic kidney disease (ADPKD) is heterogeneous with regard to genic and allelic heterogeneity, as well as phenotypic variability. The genotype-phenotype relationship in ADPKD is not completely understood. Here, we studied 741 patients with ADPKD from 519 pedigrees in the Genkyst cohort and confirmed that renal survival associated with PKD2 mutations was approximately 20 years longer than that associated with PKD1 mutations. The median age at onset of ESRD was 58 years for PKD1 carriers and 79 years for PKD2 carriers. Regarding the allelic effect on phenotype, in contrast to previous studies, we found that the type of PKD1 mutation, but not its position, correlated strongly with renal survival. The median age at onset of ESRD was 55 years for carriers of a truncating mutation and 67 years for carriers of a nontruncating mutation.
    [Show full text]
  • Human Gene Copy Number Spectra Analysis in Congenital Heart Malformations Aoy Tomita-Mitchell Medical College of Wisconsin
    CORE Metadata, citation and similar papers at core.ac.uk Provided by epublications@Marquette Marquette University e-Publications@Marquette Mathematics, Statistics and Computer Science Mathematics, Statistics and Computer Science, Faculty Research and Publications Department of 5-1-2012 Human gene copy number spectra analysis in congenital heart malformations Aoy Tomita-Mitchell Medical College of Wisconsin Donna K. Mahnke Medical College of Wisconsin Craig Struble Marquette University, [email protected] Maureen E. Tuffnell Marquette University Karl D. Stamm Marquette University See next page for additional authors Accepted version. Physiological Genomics, Vol. 44, No. 9 (May 2012): 518-541. DOI. © 2012 The American Physiological Society. Used with permission. Authors Aoy Tomita-Mitchell, Donna K. Mahnke, Craig Struble, Maureen E. Tuffnell, Karl D. Stamm, Mats Hidestrand, Susan Harris, Mary A. Goetsch, Pippa Simpson, David P. Bick, Ulrich Broeckel, Andrew N. Pelech, James S. Tweddell, and Michael Mitchell This article is available at e-Publications@Marquette: https://epublications.marquette.edu/mscs_fac/272 NOT THE PUBLISHED VERSION; this is the author’s final, peer-reviewed manuscript. The published version may be accessed by following the link in the citation at the bottom of the page. Human Gene Copy Number Spectra Analysis in Congenital Heart Malformations Aoy Tomita-Mitchell Department of Surgery, Division of Cardiothoracic Surgery; Biotechnology and Bioengineering Center; Human and Molecular Genetics Center; Medical College of Wisconsin; Milwaukee, WI Donna K. Mahnke Department of Surgery, Division of Cardiothoracic Surgery; Biotechnology and Bioengineering Center; Human and Molecular Genetics Center; Medical College of Wisconsin; Milwaukee, WI Craig A. Struble Department of Mathematics, Statistics and Computer Science; Marquette University; Milwaukee, WI Maureen E.
    [Show full text]
  • Changes in Expression of P2X1 Receptors and Connexin 43 in the Rat Myometrium During Pregnancy
    Changes in expression of P2X1 receptors and connexin 43 in the rat myometrium during pregnancy Tina Khanam, B.Sc., and Geoffrey Burnstock, Ph.D., D.Sc. Autonomic Neuroscience Centre, Royal Free and University College Medical School, London, United Kingdom Objective: To investigate the expression of P2X1 receptors and connexin 43 in gap junctions between smooth mus- cle cells. Contraction mediated by P2X receptors is known to occur in the bladder and male reproductive tract, and cell–cell coupling of smooth muscle via gap junctions is essential for synchronized rhythmic activity of these tissues. Design: We selected for this study rat myometrial smooth muscle during pregnancy and at postpartum day l. Setting: University medical school. Animal(s): Laboratory rats. Intervention(s): Rats were mated and became pregnant. Main Outcome Measure(s): Immunostaining and fluorescence and confocal microscopy. Result(s): The level of P2X1 receptor expression remained low throughout pregnancy (days 4 to 20) but was greatly up-regulated at day 22 (postpartum day 1). Connexin 43 expression showed a pattern of up-regulation, with progression through pregnancy and peaking near labor, but exhibited a rapid down-regulation after parturition. Conclusion(s): The functional significance of the changes in connexin 43 and P2X1 receptor expression that have been observed is discussed in relation to triggering and modulation of uterine contractility during and after preg- nancy. (Fertil SterilÒ 2007;88(Suppl 2):1174–9. Ó2007 by American Society for Reproductive Medicine.) Key Words: P2X1 receptor, connexin 43, myometrium, rat, confocal microscopy, immunofluorescence A recent article has shown that P2X1 receptors are closely as- connexins and P2 receptor-mediated processes.
    [Show full text]
  • Supplementary Materials
    Supplementary materials Supplementary Table S1: MGNC compound library Ingredien Molecule Caco- Mol ID MW AlogP OB (%) BBB DL FASA- HL t Name Name 2 shengdi MOL012254 campesterol 400.8 7.63 37.58 1.34 0.98 0.7 0.21 20.2 shengdi MOL000519 coniferin 314.4 3.16 31.11 0.42 -0.2 0.3 0.27 74.6 beta- shengdi MOL000359 414.8 8.08 36.91 1.32 0.99 0.8 0.23 20.2 sitosterol pachymic shengdi MOL000289 528.9 6.54 33.63 0.1 -0.6 0.8 0 9.27 acid Poricoic acid shengdi MOL000291 484.7 5.64 30.52 -0.08 -0.9 0.8 0 8.67 B Chrysanthem shengdi MOL004492 585 8.24 38.72 0.51 -1 0.6 0.3 17.5 axanthin 20- shengdi MOL011455 Hexadecano 418.6 1.91 32.7 -0.24 -0.4 0.7 0.29 104 ylingenol huanglian MOL001454 berberine 336.4 3.45 36.86 1.24 0.57 0.8 0.19 6.57 huanglian MOL013352 Obacunone 454.6 2.68 43.29 0.01 -0.4 0.8 0.31 -13 huanglian MOL002894 berberrubine 322.4 3.2 35.74 1.07 0.17 0.7 0.24 6.46 huanglian MOL002897 epiberberine 336.4 3.45 43.09 1.17 0.4 0.8 0.19 6.1 huanglian MOL002903 (R)-Canadine 339.4 3.4 55.37 1.04 0.57 0.8 0.2 6.41 huanglian MOL002904 Berlambine 351.4 2.49 36.68 0.97 0.17 0.8 0.28 7.33 Corchorosid huanglian MOL002907 404.6 1.34 105 -0.91 -1.3 0.8 0.29 6.68 e A_qt Magnogrand huanglian MOL000622 266.4 1.18 63.71 0.02 -0.2 0.2 0.3 3.17 iolide huanglian MOL000762 Palmidin A 510.5 4.52 35.36 -0.38 -1.5 0.7 0.39 33.2 huanglian MOL000785 palmatine 352.4 3.65 64.6 1.33 0.37 0.7 0.13 2.25 huanglian MOL000098 quercetin 302.3 1.5 46.43 0.05 -0.8 0.3 0.38 14.4 huanglian MOL001458 coptisine 320.3 3.25 30.67 1.21 0.32 0.9 0.26 9.33 huanglian MOL002668 Worenine
    [Show full text]
  • Draft COMP Agenda 16-18 January 2018
    12 January 2018 EMA/COMP/818236/2017 Inspections, Human Medicines Pharmacovigilance and Committees Committee for Orphan Medicinal Products (COMP) Draft agenda for the meeting on 16-18 January 2018 Chair: Bruno Sepodes – Vice-Chair: Lesley Greene 16 January 2018, 09:00-19:30, room 2F 17 January 2018, 08:30-19:30, room 2F 18 January 2018, 08:30-18:30, room 2F Health and safety information In accordance with the Agency’s health and safety policy, delegates are to be briefed on health, safety and emergency information and procedures prior to the start of the meeting. Disclaimers Some of the information contained in this agenda is considered commercially confidential or sensitive and therefore not disclosed. With regard to intended therapeutic indications or procedure scopes listed against products, it must be noted that these may not reflect the full wording proposed by applicants and may also vary during the course of the review. Additional details on some of these procedures will be published in the COMP meeting reports once the procedures are finalised. Of note, this agenda is a working document primarily designed for COMP members and the work the Committee undertakes. Note on access to documents Some documents mentioned in the agenda cannot be released at present following a request for access to documents within the framework of Regulation (EC) No 1049/2001 as they are subject to on- going procedures for which a final decision has not yet been adopted. They will become public when adopted or considered public according to the principles stated in the Agency policy on access to documents (EMA/127362/2006).
    [Show full text]
  • Genetic Determinants of Non-Syndromic Enlarged Vestibular Aqueduct: a Review
    Review Genetic Determinants of Non-Syndromic Enlarged Vestibular Aqueduct: A Review Sebastian Roesch 1 , Gerd Rasp 1, Antonio Sarikas 2 and Silvia Dossena 2,* 1 Department of Otorhinolaryngology, Head and Neck Surgery, Paracelsus Medical University, 5020 Salzburg, Austria; [email protected] (S.R.); [email protected] (G.R.) 2 Institute of Pharmacology and Toxicology, Paracelsus Medical University, 5020 Salzburg, Austria; [email protected] * Correspondence: [email protected]; Tel.: +43-(0)662-2420-80564 Abstract: Hearing loss is the most common sensorial deficit in humans and one of the most common birth defects. In developed countries, at least 60% of cases of hearing loss are of genetic origin and may arise from pathogenic sequence alterations in one of more than 300 genes known to be involved in the hearing function. Hearing loss of genetic origin is frequently associated with inner ear malformations; of these, the most commonly detected is the enlarged vestibular aqueduct (EVA). EVA may be associated to other cochleovestibular malformations, such as cochlear incomplete partitions, and can be found in syndromic as well as non-syndromic forms of hearing loss. Genes that have been linked to non-syndromic EVA are SLC26A4, GJB2, FOXI1, KCNJ10, and POU3F4. SLC26A4 and FOXI1 are also involved in determining syndromic forms of hearing loss with EVA, which are Pendred syndrome and distal renal tubular acidosis with deafness, respectively. In Caucasian cohorts, approximately 50% of cases of non-syndromic EVA are linked to SLC26A4 and a large fraction of Citation: Roesch, S.; Rasp, G.; patients remain undiagnosed, thus providing a strong imperative to further explore the etiology of Sarikas, A.; Dossena, S.
    [Show full text]