DOCSLIB.ORG

Integrin Α8 and Pcdh15 Act As a Complex to Regulate Cilia Biogenesis in Sensory Cells Linda Goodman and Marisa Zallocchi*

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Integrin Α8 and Pcdh15 Act As a Complex to Regulate Cilia Biogenesis in Sensory Cells Linda Goodman and Marisa Zallocchi* © 2017. Published by The Company of Biologists Ltd | Journal of Cell Science (2017) 130, 3698-3712 doi:10.1242/jcs.206201 RESEARCH ARTICLE Integrin α8 and Pcdh15 act as a complex to regulate cilia biogenesis in sensory cells Linda Goodman and Marisa Zallocchi* ABSTRACT sensory neuron physiology (hearing/balance, olfaction and vision), The way an organism perceives its surroundings depends on sensory as well as organ development and function (Delmaghani et al., systems and the highly specialized cilia present in the neurosensory 2016; Grati et al., 2015; Jagger et al., 2011; Rachel et al., 2012). cells. Here, we describe the existence of an integrin α8 (Itga8) and Hair cells, the neurosensory cells of the auditory and vestibular protocadherin-15a (Pcdh15a) ciliary complex in neuromast hair cells systems, are the mechanosensors for the perception of sound and in a zebrafish model. Depletion of the complex via downregulation or head movements. Projecting from their apical surface is the hair loss-of-function mutation leads to a dysregulation of cilia biogenesis bundle, which consists of rows of ascending height actin-filled and endocytosis. At the molecular level, removal of the complex stereocilia tethered to a single primary cilium, the kinocilium blocks the access of Rab8a into the cilia as well as normal recruitment (Cosgrove and Zallocchi, 2014). During hair bundle development of ciliary cargo by centriolar satellites. These defects can be reversed the kinocilium is physically connected via extracellular linkages by the introduction of a constitutively active form of Rhoa, suggesting formed between cadherin-23 (Cdh23) and protocadherin-15 that Itga8–Pcdh15a complex mediates its effect through the (Pcdh15), which are essential for proper hair bundle integrity and activation of this small GTPase and probably by the regulation of function (Kazmierczak et al., 2007; Webb et al., 2011). Mutations in actin cytoskeleton dynamics. Our data points to a novel mechanism CDH23 or PCDH15 are associated with Usher syndrome type I and involved in the regulation of sensory cilia development, with the non-syndromic hearing loss in humans (Cosgrove and Zallocchi, corresponding implications for normal sensory function. 2014). Although the kinocilium is important for the establishment of hair bundle polarity (Jones and Chen, 2008), it may also play KEY WORDS: Sensory cilia, Usher syndrome, Pcdh15, Integrin α8 additional roles as a modulator of mechanotransduction activity in immature hair cells as well as a linkage coupling the hair cell bundle INTRODUCTION to components of the extracellular matrix (ECM) (Roberts et al., Non-motile or primary cilia are centriole-derived, microtubule- 1988; Kindt et al., 2012). Recently, it has been shown that mutations based projections present in most metazoan cell types that are in two kinociliary proteins, Cdc14a and Dcdc2, are associated with involved in sensory processes such as mechanosensation, human recessive deafness (Delmaghani et al., 2016; Grati et al., chemosensation and photosensation (Leroux, 2007; Falk et al., 2015). Zebrafish morphants (MOs) for dcdc2 and cdc14a showed 2015). The formation of cilia includes the assembly of the axoneme kinocilium abnormalities with the concomitant defects in hair cell and the directional transport of ciliary proteins by membranous and morphology and function, reinforcing the notion of a direct non-membranous trafficking (Nachury et al., 2007; Westlake et al., involvement of primary cilia in hair cell function (Delmaghani 2011). In recent years significant progress has been made in the et al., 2016; Grati et al., 2015). identification of components involved in vesicular trafficking Integrins are heterodimeric cell surface receptors composed of α necessary for primary ciliogenesis (Leroux, 2007; Pazour and and β subunits that function as adhesion molecules by binding Bloodgood, 2008). Among the proteins identified, Rab8a, a extracellular matrix (ECM) proteins and as receptors by mediating member of the Rab family of small GTPases, plays a critical signal transduction (Müller et al., 1997). In particular, integrin α8 function (Deretic et al., 1995; Nachury et al., 2007). To promote (Itga8) has an obligatory association with the β1 subunit (Itgb1; cilia biogenesis, Rab8 needs to be activated by its guanine Müller et al., 1997) and is selectively incorporated into the apical nucleotide exchange factor, Rabin8 (also known as RAB3IP), membrane of hair cells during development where it is thought to which is targeted to the base of the cilia by GTP-Rab11-positive initiate the assembly of transmembrane complexes necessary for the vesicles (Nachury et al., 2007; Westlake et al., 2011; Hehnly et al., maturation of apical structures (Littlewood-Evans and Müller, 2012). In vertebrates, primary cilia are implicated in several 2000). Itga8-deficient mice have balance problems, abnormal developmental pathways and act as signaling centers mediating stereocilia and fusions between stereocilia and kinocilium, with intercellular communications (Ezratty et al., 2011; Breunig et al., perturbations in the distribution of ECM components. At the 2008; Delling et al., 2016). Dysfunction of cilia, due to mutations in molecular level, Itga8 modulates actin stress fiber assembly via a ciliary proteins or proteins involved in vesicular transport, is Rhoa-dependent mechanism (Zargham et al., 2007a,b; Benoit associated with a broad spectrum of human disorders that affect et al., 2009). Mutations in Itga8 have been associated with bilateral renal agenesis and Fraser syndrome (Humbert et al., 2014; Talbot et al., 2016). Center for Sensory Neuroscience, Boys Town National Research Hospital, Omaha, NE 68131, USA. The present work focuses on the kinocilium of neuromast hair cells. Using zebrafish as the experimental model, we demonstrated *Author for correspondence ([email protected]) ciliary localization and an association between Itga8 and Pcdh15. L.G., 0000-0001-9564-6426; M.Z., 0000-0002-6520-988X Loss of Itga8 or Pcdh15 function leads to a common phenotype, including kinociliary length dysregulation, impairment of Received 15 May 2017; Accepted 1 September 2017 endocytosis, and Rab8 and centrin mislocalization. These defects Journal of Cell Science 3698 RESEARCH ARTICLE Journal of Cell Science (2017) 130, 3698-3712 doi:10.1242/jcs.206201 can be explained by a reduction in Rhoa activation, since The number of cells harboring a kinocilium was quantified in constitutively active Rhoa is able to rescue these defects in Itga8 MOs and orbiter mutants (Fig. S2A). No significant differences and Pcdh15a knockdown and mutant zebrafish. were observed at 3 dpf. However, at 5 dpf orbiters showed a significant reduction (∼30–40%) in the fraction of cells carrying a RESULTS cilium compared to that in wild-type (WT) animals. Since the Absence of Itga8 or Pcdh15a affects kinocilia elongation number of mature hair cells per neuromast was similar across and/or maintenance treatments, and because we did not observe any increase in hair cell Preliminary results from our laboratory performed in mouse death as judged with a TUNEL assay (Fig. S2B–E), these results auditory hair cells suggested the existence of a functional Itga8– suggest that Pcdh15a and Itga8 proteins are not only required for Pcdh15 complex. To extend these findings in a more suitable model, efficient cilia elongation in neurosensory cells but also for proper we decided to analyze whether defects in Itga8 or Pcdh15a proteins cilia maintenance. resulted in zebrafish hair cell abnormalities. Knockdown zebrafish To confirm the activation state of Rhoa, rhotekin pulldown and for both of these proteins were generated for by the injection of sub- immunofluorescence experiments were performed in MOs and optimal doses of morpholino suspensions into one-cell stage eggs mutants (Fig. 2; Fig. S3). Total lysates from 1–2 dpf larvae were (hereafter denoted MOs), and analyzed at 3 days post fertilization prepared from controls, Itga8 MOs, Pcdh15a MOs and the (dpf). When studying their gross morphology, ∼30% of the MOs corresponding rescued MOs (+cRNA), and Rhoa activation (Itga8 or Pcdh15a) showed pericardial edema and slight body analyzed by pulldown assay (Fig. 2A). We observed a significant curvature (Fig. S1A–E′ and Table S1). Since these defects were not decrease in active Rhoa abundance (GTP-Rhoa, framed area in observed in the pcdh15a zebrafish mutant lines (orbiters, Seiler Fig. 2A–C) when Itga8 or Pcdh15a were knocked down compared et al., 2005) (Fig. S1F–H), they were considered morpholino off- to controls. This deficiency in Rhoa activation was rescued when target effects and excluded from our experiments. Apical hair cell MOs were co-injected with the full-length cRNAs for itga8 or morphology was analyzed in control, MOs and orbiter mutants by pcdh15a, confirming the direct involvement of these proteins in co-staining for phalloidin (a hair cell bundle marker) and acetylated Rhoa regulation. The Rhoa activation state was also evaluated in the tubulin (an axoneme marker) (Fig. 1A–O). Super-resolution orbiter lines at 5 dpf (Fig. 2B). Again, we observed a significant structured illumination microscopy (SR-SIM) analysis showed a decrease in the amount of GTP-Rhoa (Fig. 2B, framed area) when significant reduction of the kinociliary length in the itga8- and comparing the pull down results from mutant and WT animals pcdh15a-deficient animals (Fig. 1B,C,I–L) compared to that in the (Fig. 2F), directly implicating Pcdh15a in Rhoa activation and at the corresponding controls (Fig. 1A,H,K,L). The orbiter mutations not same time confirming the specificity of the Pcdh15a MO only resulted in an average reduction in the kinociliary length phenotype. Finally, total lysates from 1–2 dpf larvae injected with (Fig. 1L) but also in a dysregulation of ciliogenesis in general, as the itga8 MO suspension alone or in combination with the CA rhoa determined by the broader variation of the individual ciliary lengths cRNAs were employed to validate the activation state of Rhoa (Fig. 1M) and by a shift in the distribution of the kinociliary length (Fig. 2C,G). As expected, co-injection of CA Rhoab (Itga8 MO+b) frequencies towards shorter kinocilia (Fig.
Load more

Recommended publications
  • A Clinicopathological and Molecular Genetic Analysis of Low-Grade Glioma in Adults
    A CLINICOPATHOLOGICAL AND MOLECULAR GENETIC ANALYSIS OF LOW-GRADE GLIOMA IN ADULTS Presented by ANUSHREE SINGH MSc A thesis submitted in partial fulfilment of the requirements of the University of Wolverhampton for the degree of Doctor of Philosophy Brain Tumour Research Centre Research Institute in Healthcare Sciences Faculty of Science and Engineering University of Wolverhampton November 2014 i DECLARATION This work or any part thereof has not previously been presented in any form to the University or to any other body whether for the purposes of assessment, publication or for any other purpose (unless otherwise indicated). Save for any express acknowledgments, references and/or bibliographies cited in the work, I confirm that the intellectual content of the work is the result of my own efforts and of no other person. The right of Anushree Singh to be identified as author of this work is asserted in accordance with ss.77 and 78 of the Copyright, Designs and Patents Act 1988. At this date copyright is owned by the author. Signature: Anushree Date: 30th November 2014 ii ABSTRACT The aim of the study was to identify molecular markers that can determine progression of low grade glioma. This was done using various approaches such as IDH1 and IDH2 mutation analysis, MGMT methylation analysis, copy number analysis using array comparative genomic hybridisation and identification of differentially expressed miRNAs using miRNA microarray analysis. IDH1 mutation was present at a frequency of 71% in low grade glioma and was identified as an independent marker for improved OS in a multivariate analysis, which confirms the previous findings in low grade glioma studies.
    [Show full text]
  • Supplementary Table 1: Adhesion Genes Data Set
    Supplementary Table 1: Adhesion genes data set PROBE Entrez Gene ID Celera Gene ID Gene_Symbol Gene_Name 160832 1 hCG201364.3 A1BG alpha-1-B glycoprotein 223658 1 hCG201364.3 A1BG alpha-1-B glycoprotein 212988 102 hCG40040.3 ADAM10 ADAM metallopeptidase domain 10 133411 4185 hCG28232.2 ADAM11 ADAM metallopeptidase domain 11 110695 8038 hCG40937.4 ADAM12 ADAM metallopeptidase domain 12 (meltrin alpha) 195222 8038 hCG40937.4 ADAM12 ADAM metallopeptidase domain 12 (meltrin alpha) 165344 8751 hCG20021.3 ADAM15 ADAM metallopeptidase domain 15 (metargidin) 189065 6868 null ADAM17 ADAM metallopeptidase domain 17 (tumor necrosis factor, alpha, converting enzyme) 108119 8728 hCG15398.4 ADAM19 ADAM metallopeptidase domain 19 (meltrin beta) 117763 8748 hCG20675.3 ADAM20 ADAM metallopeptidase domain 20 126448 8747 hCG1785634.2 ADAM21 ADAM metallopeptidase domain 21 208981 8747 hCG1785634.2|hCG2042897 ADAM21 ADAM metallopeptidase domain 21 180903 53616 hCG17212.4 ADAM22 ADAM metallopeptidase domain 22 177272 8745 hCG1811623.1 ADAM23 ADAM metallopeptidase domain 23 102384 10863 hCG1818505.1 ADAM28 ADAM metallopeptidase domain 28 119968 11086 hCG1786734.2 ADAM29 ADAM metallopeptidase domain 29 205542 11085 hCG1997196.1 ADAM30 ADAM metallopeptidase domain 30 148417 80332 hCG39255.4 ADAM33 ADAM metallopeptidase domain 33 140492 8756 hCG1789002.2 ADAM7 ADAM metallopeptidase domain 7 122603 101 hCG1816947.1 ADAM8 ADAM metallopeptidase domain 8 183965 8754 hCG1996391 ADAM9 ADAM metallopeptidase domain 9 (meltrin gamma) 129974 27299 hCG15447.3 ADAMDEC1 ADAM-like,
    [Show full text]
  • TMHS Is an Integral Component of the Mechanotransduction Machinery of Cochlear Hair Cells
    TMHS Is an Integral Component of the Mechanotransduction Machinery of Cochlear Hair Cells Wei Xiong,1 Nicolas Grillet,1 Heather M. Elledge,1 Thomas F.J. Wagner,1 Bo Zhao,1 Kenneth R. Johnson,2 Piotr Kazmierczak,1 and Ulrich Mu¨ller1,* 1The Dorris Neuroscience Center, Department of Cell Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA 2The Jackson Laboratory, Bar Harbor, ME 04609, USA *Correspondence: [email protected] http://dx.doi.org/10.1016/j.cell.2012.10.041 SUMMARY deflection of the stereociliary bundles, which directly control the activity of the mechanotransduction channels in stereocilia. Hair cells are mechanosensors for the perception of It is thought that tip links, fine extracellular filaments that connect sound, acceleration, and fluid motion. Mechano- the tips of neighboring stereocilia, transmit tension force onto transduction channels in hair cells are gated by tip the transduction channels (Gillespie and Mu¨ ller, 2009). links, which connect the stereocilia of a hair cell in In recent years, significant progress has been made in the direction of their mechanical sensitivity. The the identification of components of the mechanotransduction molecular constituents of the mechanotransduction machinery of hair cells (Figure 1A). These studies have shown that tip links are formed by CDH23 homodimers that interact channels of hair cells are not known. Here, we show with PCDH15 homodimers to form the upper and lower parts that mechanotransduction is impaired in mice lack- of tip links (Ahmed et al., 2006; Kazmierczak et al., 2007; ing the tetraspan TMHS. TMHS binds to the tip-link Siemens et al., 2004; So¨ llner et al., 2004).
    [Show full text]
  • 1 Structural Studies of a Protocadherin-15 Fragment Essential for Hearing
    Structural Studies of a Protocadherin-15 Fragment Essential for Hearing A thesis presented By Conghui Chen To The Committee on Degrees in Chemistry ad Biochemistry In partial fulfillment of the requirements For a degree of Bachelor of Science with Research Distinction In the field of Biochemistry Research Advisor: Dr. Marcos Sotomayor, Assistant Professor of Department of Chemistry and Biochemistry Defense Committee: Dr. John Shimko, Chemistry Lecturer of Department of Chemistry and Biochemistry The Ohio State University Columbus, Ohio April 13th, 2016 1 Statement of Research I conducted the research presented in this thesis under the professional guidance of Dr. Marcos Sotomayor of The Ohio State University Main Campus Chemistry and Biochemistry Department. I joined the Sotomayor lab in August of 2013, during my second year at the university. I was trained in the process of protein purification and cell culture by visiting graduate student Deryanur Kilic in conjunction with Dr. Sotomayor. Dr. Sotomayor offered knowledgeable tutelage in designing and performing the experiments, analyzing data, and the writing of this thesis. All molecular modeling and analysis was performed with the guidance and assistance from Dr. Marcos Sotomayor and Dr. Raul Araya-Secchi. My research was generously funded by The Ohio State University Chemistry and Biochemistry Undergraduate Research Scholarship from autumn 2013 until spring 2015. I performed research as part of the Biochemistry 4998 and 4999 courses as a requirement for the completion of the thesis. 2 Abstract Sound travels through the external and middle ear to the fluid-filled cochlea where mechanosensitive hair cells transform it into electrochemical signals. On the apical side of each hair cell, a set of hair-like protrusions, called stereocilia form a bundle with filamentous connections (tip links) that are essential for hearing.
    [Show full text]
  • Tip-Link Protein Protocadherin 15 Interacts with Transmembrane Channel-Like Proteins TMC1 and TMC2
    Tip-link protein protocadherin 15 interacts with transmembrane channel-like proteins TMC1 and TMC2 Reo Maedaa,b,1, Katie S. Kindta,b,1,2, Weike Moa,b,1, Clive P. Morgana,b, Timothy Ericksona,b, Hongyu Zhaoa,b, Rachel Clemens-Grishama,b, Peter G. Barr-Gillespiea,b, and Teresa Nicolsona,b,3 aOregon Hearing Research Center and bVollum Institute, Oregon Health and Science University, Portland, OR 97239 Edited by A. J. Hudspeth, Howard Hughes Medical Institute, The Rockefeller University, New York, NY, and approved July 23, 2014 (received for review February 3, 2014) The tip link protein protocadherin 15 (PCDH15) is a central compo- vestibular deficits, along with the complete absence of normal nent of the mechanotransduction complex in auditory and vestib- mechanotransduction currents in auditory and vestibular hair cells ular hair cells. PCDH15 is hypothesized to relay external forces to the (17). Changes in calcium permeability through the transduction mechanically gated channel located near its cytoplasmic C terminus. channel of cochlear hair cells were observed for Tmc1 Tmc2 How PCDH15 is coupled to the transduction machinery is not clear. double-mutant mice, as well as in single mutants of either gene Using a membrane-based two-hybrid screen to identify proteins (10, 18, 19). In further support of the idea that TMCs are pore- that bind to PCDH15, we detected an interaction between zebrafish forming subunits of the transduction channel, mouse vestibular Pcdh15a and an N-terminal fragment of transmembrane channel- hair cells that express only the dominant Beethoven (M412K) al- like 2a (Tmc2a). Tmc2a is an ortholog of mammalian TMC2, which lele of Tmc1, in the absence of any wild-type TMC1 or TMC2, along with TMC1 has been implicated in mechanotransduction in display altered single-channel transduction currents (10).
    [Show full text]
  • Cadherins As Targets for Genetic Diseases
    Downloaded from http://cshperspectives.cshlp.org/ on September 28, 2021 - Published by Cold Spring Harbor Laboratory Press Cadherins as Targets for Genetic Diseases Aziz El-Amraoui1,2,3 and Christine Petit1,2,3,4 1Institut Pasteur, Unite´ de Ge´ne´tique et Physiologie de l’Audition, 25 rue du Dr Roux, 75015 Paris, France 2INSERM UMRS587, 75015 Paris, France 3UPMC, F75015 Paris, France 4Colle`ge de France, 75005 Paris, France Correspondence: [email protected], [email protected] The 6-billion human population provides a vast reservoir of mutations, which, in addition to the opportunity of detecting very subtle defects, including specific cognitive dysfunctions as well as late appearing disorders, offers a unique background in which to investigate the roles of cell–cell adhesion proteins. Here we focus on inherited human disorders involving members of the cadherin superfamily. Most of the advances concern monogenic disorders. Yet,with the development of single nucleotide polymorphism (SNP) association studies, cad- herin genes are emerging as susceptibility genes in multifactorial disorders. Various skin and heart disorders revealed the critical role played by desmosomal cadherins in epidermis, hairs, and myocardium, which experience high mechanical stress. Of particular interest in that respect is the study of Usher syndrome type 1 (USH1), a hereditary syndromic form of deafness. Studies of USH1 brought to light the crucial role of transient fibrous links formed by cadherin 23 and protocadherin 15 in the cohesion of the developing hair bundle, the mechanoreceptive structure of the auditory sensory cells, as well as the involvement of these cadherins in the formation of the tip-link, a key component of the mechano-electrical transduction machinery.
    [Show full text]
  • Identification and the Significance of Selective Proteins in Bile And
    University of Arkansas, Fayetteville ScholarWorks@UARK Theses and Dissertations 12-2014 Identification and the Significance of Selective Proteins in Bile and Plasma of Normal and Health- compromised Chickens Balamurugan Packialakshmi University of Arkansas, Fayetteville Follow this and additional works at: http://scholarworks.uark.edu/etd Part of the Analytical Chemistry Commons, Animal Diseases Commons, and the Poultry or Avian Science Commons Recommended Citation Packialakshmi, Balamurugan, "Identification and the Significance of Selective Proteins in Bile and Plasma of Normal and Health- compromised Chickens" (2014). Theses and Dissertations. 2083. http://scholarworks.uark.edu/etd/2083 This Dissertation is brought to you for free and open access by ScholarWorks@UARK. It has been accepted for inclusion in Theses and Dissertations by an authorized administrator of ScholarWorks@UARK. For more information, please contact [email protected], [email protected]. Identification and the Significance of Selective Proteins in Bile and Plasma of Normal and Health-compromised Chickens Identification and the Significance of Selective Proteins in Bile and Plasma of Normal and Health-compromised Chickens A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Cell and Molecular Biology by Balamurugan Packialakshmi Tamilnadu Agricultural University, Bachelor of Technology in Agricultural Biotechnology, 2007 Govind Ballabh Pant University of Agriculture and Technology, Master of Science in Molecular Biology and Biotechnology, 2009 December 2014 University of Arkansas This dissertation is approved for the recommendation to the graduate council. _____________________ Dr. Narayan. C. Rath Dissertation Director _____________________ _____________________ Dr. Jackson O. Lay, Jr. Dr. Robert F. Wideman, Jr. Committee member Committee member ____________________ Dr.
    [Show full text]
  • The NIH Catalyst Spoke with Copeland Chief Se- and Jenkins— and CONTENTS Nior Investigator, Respectively, of the Bill Branson
    Fostering Communication and Collaboration The nihCatalyst A Publication for NIH Intramural Scientists National Institutes of Health bOffice of the Director' e Volume 11, Issue 6* November-December 2003 CC Celebrates 50th at Research Festival > the Sum of the Parts Giants Standing Copeland and Jenkins On the Shoulders of Giants And the Development Of Mouse Cancer Genetics by Fran Pollner DeVita it by PeterJ. Kozel ince put this way: “It’s nice V to come home.” And in one way or an- other, each of the speak- ers who took the stage to commemorate the CC’s 50th anniversary ex- pressed an affection for the daily working envi- ronment at NIFI and its research hospital that Bill Branson conjured up the image of CC DirectorJohn Gallin salutes former and current NIH NancyJenkins Neal Copeland home: scientists and a half-century of transforming research t’s hard to say what the NIH sci- The collaborative entific directors were honoring and feisty spirit among colleagues remi- The perseverance, warmth, and mu- I when they picked husband-and- niscent of the best kind of sibling cama- tual regard that characterized the rela- wife cancer genetics investigators raderie—and squabbling tionships between the physician-re- Neal Copeland and Nancy Jenkins searchers and the patients undergoing to present this year’s Mider Lecture. experimental treatments, sometimes ex- Was it their legacy—almost 700 pa- tending through decades of follow-up pers and the reference mouse ge- The culture of NIFf, like that in a nome map? Was it their astonishing nurturing family, that supported the pur- record of training successful scien- suit of new ideas and personal intellec- tists? Was it their innovative “recom- tual expansion bineering” technique that’s revolu- They all saw the past—studded with tionizing the manipulation of DNA? the gems of biomedical research that Or was it their almost unparalleled, contributed enormously to science and highly productive collaboration, now human health—as prologue to the fu- in its third decade? ture.
    [Show full text]
  • The Alter Retina: Alternative Splicing of Retinal Genes in Health and Disease
    International Journal of Molecular Sciences Review The Alter Retina: Alternative Splicing of Retinal Genes in Health and Disease Izarbe Aísa-Marín 1,2 , Rocío García-Arroyo 1,3 , Serena Mirra 1,2 and Gemma Marfany 1,2,3,* 1 Departament of Genetics, Microbiology and Statistics, Avda. Diagonal 643, Universitat de Barcelona, 08028 Barcelona, Spain; [email protected] (I.A.-M.); [email protected] (R.G.-A.); [email protected] (S.M.) 2 Centro de Investigación Biomédica en Red Enfermedades Raras (CIBERER), Instituto de Salud Carlos III (ISCIII), Universitat de Barcelona, 08028 Barcelona, Spain 3 Institute of Biomedicine (IBUB, IBUB-IRSJD), Universitat de Barcelona, 08028 Barcelona, Spain * Correspondence: [email protected] Abstract: Alternative splicing of mRNA is an essential mechanism to regulate and increase the diversity of the transcriptome and proteome. Alternative splicing frequently occurs in a tissue- or time-specific manner, contributing to differential gene expression between cell types during development. Neural tissues present extremely complex splicing programs and display the highest number of alternative splicing events. As an extension of the central nervous system, the retina constitutes an excellent system to illustrate the high diversity of neural transcripts. The retina expresses retinal specific splicing factors and produces a large number of alternative transcripts, including exclusive tissue-specific exons, which require an exquisite regulation. In fact, a current challenge in the genetic diagnosis of inherited retinal diseases stems from the lack of information regarding alternative splicing of retinal genes, as a considerable percentage of mutations alter splicing Citation: Aísa-Marín, I.; or the relative production of alternative transcripts. Modulation of alternative splicing in the retina García-Arroyo, R.; Mirra, S.; Marfany, is also instrumental in the design of novel therapeutic approaches for retinal dystrophies, since it G.
    [Show full text]
  • Dynamic Changes of Muscle Insulin Sensitivity After Metabolic Surgery
    ARTICLE https://doi.org/10.1038/s41467-019-12081-0 OPEN Dynamic changes of muscle insulin sensitivity after metabolic surgery Sofiya Gancheva1,2,3,11, Meriem Ouni 3,4,11, Tomas Jelenik2,3, Chrysi Koliaki1,2,3,5, Julia Szendroedi1,2,3, Frederico G.S. Toledo6, Daniel F. Markgraf 2,3, Dominik H. Pesta 2,3, Lucia Mastrototaro2,3, Elisabetta De Filippo2,3, Christian Herder 1,2,3, Markus Jähnert 3,4, Jürgen Weiss3,7, Klaus Strassburger 3,8, Matthias Schlensak9, Annette Schürmann 3,4,10,12 & Michael Roden 1,2,3,12 1234567890():,; The mechanisms underlying improved insulin sensitivity after surgically-induced weight loss are still unclear. We monitored skeletal muscle metabolism in obese individuals before and over 52 weeks after metabolic surgery. Initial weight loss occurs in parallel with a decrease in muscle oxidative capacity and respiratory control ratio. Persistent elevation of intramyocel- lular lipid intermediates, likely resulting from unrestrained adipose tissue lipolysis, accom- panies the lack of rapid changes in insulin sensitivity. Simultaneously, alterations in skeletal muscle expression of genes involved in calcium/lipid metabolism and mitochondrial function associate with subsequent distinct DNA methylation patterns at 52 weeks after surgery. Thus, initial unfavorable metabolic changes including insulin resistance of adipose tissue and skeletal muscle precede epigenetic modifications of genes involved in muscle energy metabolism and the long-term improvement of insulin sensitivity. 1 Division of Endocrinology and Diabetology, Medical Faculty, Heinrich-Heine University, Düsseldorf, Germany. 2 Institute for Clinical Diabetology, German Diabetes Center, Leibniz Center for Diabetes Research, Heinrich Heine University, Düsseldorf, Germany. 3 German Center for Diabetes Research (DZD e.V.), Neuherberg, Germany.
    [Show full text]
  • Species-Dependent Heterophilic and Homophilic Cadherin Interactions In
    bioRxiv preprint doi: https://doi.org/10.1101/2020.09.01.278846; this version posted September 2, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 1 SPECIES-DEPENDENT HETEROPHILIC AND HOMOPHILIC CADHERIN INTERACTIONS IN 2 INTESTINAL INTERMICROVILLAR LINKS 3 4 Michelle E. Gray*1, Zachary R. Johnson*§2, Debadrita Modak*2, Matthew J. Tyska3, and Marcos Sotomayor#1,2 5 1Ohio State Biochemistry Program 6 2Department of Chemistry and Biochemistry 7 The Ohio State University, 484 W 12th Avenue, Columbus, OH 43210 8 3Department of Cell and Developmental Biology 9 Vanderbilt University School of Medicine, Nashville, TN 37232 10 *Co-first authors 11 §Current address: Marshall University, Joan C. Edwards School of Medicine, Huntington, WV 25701 12 # Corresponding Author 13 E-mail: [email protected] 14 1 bioRxiv preprint doi: https://doi.org/10.1101/2020.09.01.278846; this version posted September 2, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 1 ABSTRACT 2 Enterocytes are specialized epithelial cells lining the luminal surface of the small intestine that build densely packed 3 arrays of microvilli known as brush borders. These microvilli drive nutrient absorption and are arranged in a hexagonal 4 pattern maintained by intermicrovillar links formed by two non-classical members of the cadherin superfamily of calcium- 5 dependent cell adhesion proteins: protocadherin-24 (PCDH24, also known as CDHR2) and the mucin-like protocadherin 6 (CDHR5).
    [Show full text]
  • Deafness Mutation D572N of TMC1 Destabilizes TMC1 Expression by Disrupting LHFPL5 Binding
    Deafness mutation D572N of TMC1 destabilizes TMC1 expression by disrupting LHFPL5 binding Xiaojie Yua, Qirui Zhaoa, Xiaofen Lia, Yixuan Chenb, Ye Tiana, Shuang Liuc, Wei Xiongc, and Pingbo Huanga,b,d,e,f,1 aDivision of Life Science, Hong Kong University of Science and Technology, Hong Kong, China; bDepartment of Chemical and Biological Engineering, Hong Kong University of Science and Technology, Hong Kong, China; cSchool of Life Sciences, Tsinghua University, Beijing, China 100084; dState Key Laboratory of Molecular Neuroscience, Hong Kong University of Science and Technology, Hong Kong, China; eHong Kong University of Science and Technology (HKUST) Shenzhen Research Institute, Hong Kong University of Science and Technology, Hong Kong, China; and fHong Kong Branch of Guangdong Southern Marine Science and Engineering Laboratory (Guangzhou), Hong Kong University of Science and Technology, Hong Kong, China Edited by Robert Fettiplace, University of Wisconsin, Madison, WI, and accepted by Editorial Board Member Jeremy Nathans October 8, 2020 (received for review June 1, 2020) Transmembrane channel-like protein 1 (TMC1) and lipoma HMGIC expression is stabilized by LHFPL5 binding both in heterologous fusion partner-like 5 (LHFPL5) are recognized as two critical com- expression systems and in hair cells. Notably, we found that the ponents of the mechanotransduction complex in inner-ear hair deafness-causing mutation D572N in TMC1 disrupted TMC1- cells. However, the physical and functional interactions of TMC1 LHFPL5 interaction and destabilized TMC1 expression. and LHFPL5 remain largely unexplored. We examined the interac- tion between TMC1 and LHFPL5 by using multiple approaches, Results including our recently developed ultrasensitive microbead-based Ectopic TMC1 and LHFPL5 Physically Interact and Mutually Stabilize single-molecule pulldown (SiMPull) assay.
    [Show full text]