Petcharat Leoyklang1, Bradley Class2, Satoru Noguchi3, William A Gahl1,4, Nuria Carrillo1,2, Ichizo Nishino3, Marjan Huizing1, May Christine Malicdan1,4. 1. Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, 10 Center Drive, Bld. 10, Room 10C103 Bethesda, Maryland, 20892, USA. 2. Therapeutics for Rare and Neglected Diseases, National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, Maryland, USA. 3. Department of Neuromuscular Research, National Institute of Neuroscience, National Center of Neurology and Psychiatry, Tokyo, Japan. 4. NIH Undiagnosed Diseases Program, Common Fund, Office of the Director, National Institutes of Health, Bethesda, Maryland, USA.
Abstract
INTRODUCTION: GNE myopathy is an adult-onset muscle disorder characterized by impaired sialylation of (muscle) glycans, detectable by lectin histochemistry. We describe a standardized method to quantify (lectin-) fluorescence in muscle sections, applicable for diagnosis and response to therapy for GNE myopathy. METHODS: Muscle sections were fluorescently labeled with the sialic acid-binding Sambucus nigra agglutinin (SNA) lectin and antibodies to sarcolemma residence protein caveolin-3 (CAV-3). Entire tissue sections were imaged in tiles and fluorescence was quantified. RESULTS: SNA fluorescence co-localizing with CAV-3 was ∼50% decreased in GNE myopathy biopsies compared with muscle-matched controls, confirming previous qualitative results. DISCUSSION: This quantitative fluorescence method can accurately determine sialylation status of GNE myopathy muscle biopsies. This method is adaptable for expression of other membrane-associated muscle proteins, and may be of benefit for disorders in which therapeutic changes in expression are subtle and difficult to assess by other methods. Muscle Nerve 58: 286-292, 2018.
INTRODUCTION:GNEmyopathy is an adult-onset muscle disorder characterized by impaired sialylation of (muscle) glycans, detectable by lectin histochemistry. We describe a standardized method to quantify (lectin-) fluorescence in muscle sections, applicable for diagnosis and response to therapy for GNEmyopathy. METHODS: Muscle sections were fluorescently labeled with the sialic acid-binding Sambucus nigra agglutinin (SNA) lectin and antibodies to sarcolemma residence protein caveolin-3 (CAV-3). Entire tissue sections were imaged in tiles and fluorescence was quantified. RESULTS:SNA fluorescence co-localizing with CAV-3 was ∼50% decreased in GNEmyopathy biopsies compared with muscle-matched controls, confirming previous qualitative results. DISCUSSION: This quantitative fluorescence method can accurately determine sialylation status of GNEmyopathy muscle biopsies. This method is adaptable for expression of other membrane-associated muscle proteins, and may be of benefit for disorders in which therapeutic changes in expression are subtle and difficult to assess by other methods. Muscle Nerve 58: 286-292, 2018.
Authors: Petcharat Leoyklang; May Christine Malicdan; Tal Yardeni; Frank Celeste; Carla Ciccone; Xueli Li; Rong Jiang; William A Gahl; Nuria Carrillo-Carrasco; Miao He; Marjan Huizing Journal: Biomark Med Date: 2014 Impact factor: 2.851
Authors: E M McNally; E de Sá Moreira; D J Duggan; C G Bönnemann; M P Lisanti; H G Lidov; M Vainzof; M R Passos-Bueno; E P Hoffman; M Zatz; L M Kunkel Journal: Hum Mol Genet Date: 1998-05 Impact factor: 6.150
Authors: E Ricci; A Broccolini; T Gidaro; R Morosetti; C Gliubizzi; R Frusciante; G M Di Lella; P A Tonali; M Mirabella Journal: Neurology Date: 2006-03-14 Impact factor: 9.910
Authors: Terren K Niethamer; Tal Yardeni; Petcharat Leoyklang; Carla Ciccone; Adrian Astiz-Martinez; Katherine Jacobs; Heidi M Dorward; Patricia M Zerfas; William A Gahl; Marjan Huizing Journal: Mol Genet Metab Date: 2012-10-18 Impact factor: 4.797
Authors: C Minetti; F Sotgia; C Bruno; P Scartezzini; P Broda; M Bado; E Masetti; M Mazzocco; A Egeo; M A Donati; D Volonte; F Galbiati; G Cordone; F D Bricarelli; M P Lisanti; F Zara Journal: Nat Genet Date: 1998-04 Impact factor: 38.330