Solomon T Gizaw1, Toshiaki Koda2, Maho Amano1, Keiko Kamimura2, Tetsu Ohashi3, Hiroshi Hinou3, Shin-Ichiro Nishimura4. 1. Field of Drug Discovery Research, Faculty of Advanced Life Science, Hokkaido University, N21 W11, Sapporo 001-0021, Japan; Graduate School of Life Science, Hokkaido University, N21 W11, Sapporo 001-0021, Japan. 2. Laboratory of Embryonic and Genetic Engineering, Faculty of Advanced Life Science, Hokkaido University, N21 W11, Sapporo 001-0021, Japan. 3. Field of Drug Discovery Research, Faculty of Advanced Life Science, Hokkaido University, N21 W11, Sapporo 001-0021, Japan; Medicinal Chemistry Pharmaceuticals, Co. Ltd, N21 W12, Sapporo 001-0021, Japan; Graduate School of Life Science, Hokkaido University, N21 W11, Sapporo 001-0021, Japan. 4. Field of Drug Discovery Research, Faculty of Advanced Life Science, Hokkaido University, N21 W11, Sapporo 001-0021, Japan; Medicinal Chemistry Pharmaceuticals, Co. Ltd, N21 W12, Sapporo 001-0021, Japan; Graduate School of Life Science, Hokkaido University, N21 W11, Sapporo 001-0021, Japan. Electronic address: shin@sci.hokudai.ac.jp.
Abstract
BACKGROUND: Huntington's disease (HD) is an autosomal, dominantly inherited and progressive neurodegenerative disease, nosologically classified as the presence of intranuclear inclusion bodies and the loss of GABA-containing neurons in the neostriatum and subsequently in the cerebellar cortex. Abnormal processing of neuronal proteins can result in the misfolding of proteins and altered post-translational modification of newly synthesized proteins. Total glycomics, namely, N-glycomics, O-glycomics, and glycosphingolipidomics (GSL-omics) of HD transgenic mice would be a hallmark for central nervous system disorders in order to discover disease specific biomarkers. METHODS: Glycoblotting method, a high throughput glycomic protocol, and matrix-assisted laser desorption ionization-time of flight/mass spectrometry (MALDI-TOF/MS) were used to study the total glycome expression levels in the brain tissue (3 mice of each sex) and sera (5 mice of each sex) of HD transgenic and control mice. All experiments were performed twice and differences in the expression levels of major glycoforms were compared between HD transgenic and control mice. RESULTS: We estimated the structure and expression levels of 87 and 58N-glycans in brain tissue and sera, respectively, of HD transgenic and control mice. The present results clearly indicated that the brain glycome and their expression levels are significantly gender specific when compared with those of other tissues and serum. Core-fucosylated and bisecting-GlcNAc types of N-glycans were found in increased levels in the brain tissue HD transgenic mice. Accordingly, core-fucosylated and sialic acid (particularly N-glycolylneuraminic acid, NeuGc) for biantennary type glycans were found in increased amounts in the sera of HD transgenic mice compared to that of control mice. Core 3 type O-glycans were found in increased levels in male and in decreased levels in both the striatum and cortexes of female HD transgenic mice. Furthermore, serum levels of core 1 type O-glycans decreased and were undetected for core 2 type O-glycans for HD transgenic mice. In glycosphingolipids, GD1a in brain tissue and GM2-NeuGc serum levels were found to have increased and decreased, respectively, in HD transgenic mice compared to those of the control group mice. CONCLUSION: Total glycome expression levels are significantly different between HD transgenic and control group mice. GENERAL SIGNIFICANCE: Glycoblotting combined with MALDI-TOF/MS total glycomics warrants a comprehensive, effective, novel and versatile technique for qualitative and quantitative analysis of total glycome expression levels. Furthermore, glycome-focused studies of both environmentally and genetically rooted neurodegenerative diseases are promising candidates for the discovery of potential disease glyco-biomarkers in the post-genome era.
BACKGROUND:Huntington's disease (HD) is an autosomal, dominantly inherited and progressive neurodegenerative disease, nosologically classified as the presence of intranuclear inclusion bodies and the loss of GABA-containing neurons in the neostriatum and subsequently in the cerebellar cortex. Abnormal processing of neuronal proteins can result in the misfolding of proteins and altered post-translational modification of newly synthesized proteins. Total glycomics, namely, N-glycomics, O-glycomics, and glycosphingolipidomics (GSL-omics) of HDtransgenic mice would be a hallmark for central nervous system disorders in order to discover disease specific biomarkers. METHODS: Glycoblotting method, a high throughput glycomic protocol, and matrix-assisted laser desorption ionization-time of flight/mass spectrometry (MALDI-TOF/MS) were used to study the total glycome expression levels in the brain tissue (3 mice of each sex) and sera (5 mice of each sex) of HDtransgenic and control mice. All experiments were performed twice and differences in the expression levels of major glycoforms were compared between HDtransgenic and control mice. RESULTS: We estimated the structure and expression levels of 87 and 58N-glycans in brain tissue and sera, respectively, of HDtransgenic and control mice. The present results clearly indicated that the brain glycome and their expression levels are significantly gender specific when compared with those of other tissues and serum. Core-fucosylated and bisecting-GlcNAc types of N-glycans were found in increased levels in the brain tissue HDtransgenic mice. Accordingly, core-fucosylated and sialic acid (particularly N-glycolylneuraminic acid, NeuGc) for biantennary type glycans were found in increased amounts in the sera of HDtransgenic mice compared to that of control mice. Core 3 type O-glycans were found in increased levels in male and in decreased levels in both the striatum and cortexes of female HDtransgenic mice. Furthermore, serum levels of core 1 type O-glycans decreased and were undetected for core 2 type O-glycans for HDtransgenic mice. In glycosphingolipids, GD1a in brain tissue and GM2-NeuGc serum levels were found to have increased and decreased, respectively, in HDtransgenic mice compared to those of the control group mice. CONCLUSION: Total glycome expression levels are significantly different between HDtransgenic and control group mice. GENERAL SIGNIFICANCE: Glycoblotting combined with MALDI-TOF/MS total glycomics warrants a comprehensive, effective, novel and versatile technique for qualitative and quantitative analysis of total glycome expression levels. Furthermore, glycome-focused studies of both environmentally and genetically rooted neurodegenerative diseases are promising candidates for the discovery of potential disease glyco-biomarkers in the post-genome era.
Authors: Motamed Elsayed Mahmoud; Ibrahim F Rehan; Kh El-Dawy Ahmed; Amany Abdelrahman; Saeed Mohammadi; Ahmed F Abou-Elnaga; Mohammed Youssef; Hassan Mahmoud Diab; Doaa Salman; Asmaa Elnagar; Hesham H Mohammed; Obeid Shanab; Rawia M Ibrahim; Eslam K H Ahmed; Abd El-Latif Hesham; Arti Gupta Journal: Mol Biol Rep Date: 2019-03-26 Impact factor: 2.316