Glycoprotein Hyposialylation Gives Rise to a Nephrotic-Like Syndrome That Is Prevented by Sialic Acid Administration in GNE V572L Point-Mutant Mice
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{"title"=>"Glycoprotein hyposialylation gives rise to a nephrotic-like syndrome that is prevented by sialic acid administration in GNE V572L point-mutant mice", "type"=>"journal", "authors"=>[{"first_name"=>"Mitutoshi", "last_name"=>"Ito", "scopus_author_id"=>"36646115100"}, {"first_name"=>"Kazushi", "last_name"=>"Sugihara", "scopus_author_id"=>"8088896000"}, {"first_name"=>"Tomoya", "last_name"=>"Asaka", "scopus_author_id"=>"7005817062"}, {"first_name"=>"Tadashi", "last_name"=>"Toyama", "scopus_author_id"=>"36852885800"}, {"first_name"=>"Toru", "last_name"=>"Yoshihara", "scopus_author_id"=>"16025966500"}, {"first_name"=>"Kengo", "last_name"=>"Furuichi", "scopus_author_id"=>"7005758654"}, {"first_name"=>"Takashi", "last_name"=>"Wada", "scopus_author_id"=>"35070802300"}, {"first_name"=>"Masahide", "last_name"=>"Asano", "scopus_author_id"=>"24315446800"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"pui"=>"364092927", "doi"=>"10.1371/journal.pone.0029873", "pmid"=>"22253810", "issn"=>"19326203", "scopus"=>"2-s2.0-84855828221", "sgr"=>"84855828221"}, "id"=>"a2b481fa-ffb6-3e5f-8b37-cb46ce8fbc02", "abstract"=>"Mutations in the key enzyme of sialic acid biosynthesis, UDP-N-acetylglucosamine 2-epimerase/N-acetyl-mannosamine kinase, result in distal myopathy with rimmed vacuoles (DMRV)/hereditary inclusion body myopathy (HIBM) in humans. Sialic acid is an acidic monosaccharide that modifies non-reducing terminal carbohydrate chains on glycoproteins and glycolipids, and it plays an important role in cellular adhesions and interactions. In this study, we generated mice with a V572L point mutation in the GNE kinase domain. Unexpectedly, these mutant mice had no apparent myopathies or motor dysfunctions. However, they had a short lifespan and exhibited renal impairment with massive albuminuria. Histological analysis showed enlarged glomeruli with mesangial matrix deposition, leading to glomerulosclerosis and abnormal podocyte foot process morphologies in the kidneys. Glycan analysis using several lectins revealed glomerular epithelial cell hyposialylation, particularly the hyposialylation of podocalyxin, which is one of important molecules for the glomerular filtration barrier. Administering Neu5Ac to the mutant mice from embryonic stages significantly suppressed the albuminuria and renal pathology, and partially recovered the glomerular glycoprotein sialylation. These findings suggest that the nephrotic-like syndrome observed in these mutant mice resulted from impaired glomerular filtration due to the hyposialylation of podocyte glycoproteins, including podocalyxin. Furthermore, it was possible to prevent the nephrotic-like disease in these mice by beginning Neu5Ac treatment during gestation.", "link"=>"http://www.mendeley.com/research/glycoprotein-hyposialylation-gives-rise-nephroticlike-syndrome-prevented-sialic-acid-administration", "reader_count"=>26, "reader_count_by_academic_status"=>{"Researcher"=>6, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>4, "Student > Postgraduate"=>2, "Other"=>7, "Student > Master"=>1, "Student > Bachelor"=>3, "Professor"=>2}, "reader_count_by_user_role"=>{"Researcher"=>6, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>4, "Student > Postgraduate"=>2, "Other"=>7, "Student > Master"=>1, "Student > Bachelor"=>3, "Professor"=>2}, "reader_count_by_subject_area"=>{"Unspecified"=>2, "Biochemistry, Genetics and Molecular Biology"=>2, "Agricultural and Biological Sciences"=>13, "Medicine and Dentistry"=>3, "Neuroscience"=>1, "Pharmacology, Toxicology and Pharmaceutical Science"=>1, "Chemistry"=>3, "Immunology and Microbiology"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>3}, "Neuroscience"=>{"Neuroscience"=>1}, "Chemistry"=>{"Chemistry"=>3}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>13}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>2}, "Unspecified"=>{"Unspecified"=>2}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}}, "reader_count_by_country"=>{"Korea (South)"=>1, "United States"=>1, "Japan"=>1}, "group_count"=>0}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/692686"], "description"=>"<p>(A) PAS-stained kidney sections of ht (upper panel) and mt (lower panel) mice at 3 days after birth. Red bars indicate the nephrogenic zone. (B) Nephrogenic zone thickness compared between ht (open bars, n = 4) and mt (closed bars, n = 3) mice at 3 days after birth. (C) Relative nephron numbers compared between ht (open bars) and mt (closed bars) mice at 8 days (n = 10 ht, 9 mt) and 2 months (n = 6 ht, 6 mt) of age. Nephron number per mm<sup>2</sup> in the ht renal cortex at each age was designated as 100%. *<i>P</i><0.05 (Mann-Whitney U-test).</p>", "links"=>[], "tags"=>["nephron", "numbers"], "article_id"=>363134, "categories"=>["Biotechnology", "Biochemistry", "Marine Biology"], "users"=>["Mitutoshi Ito", "Kazushi Sugihara", "Tomoya Asaka", "Tadashi Toyama", "Toru Yoshihara", "Kengo Furuichi", "Takashi Wada", "Masahide Asano"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0029873.g004", "stats"=>{"downloads"=>1, "page_views"=>25, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Nephrogenic_zone_formation_and_nephron_numbers_in_kidneys_/363134", "title"=>"Nephrogenic zone formation and nephron numbers in kidneys.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-13 00:52:14"}
  • {"files"=>["https://ndownloader.figshare.com/files/352431", "https://ndownloader.figshare.com/files/352479", "https://ndownloader.figshare.com/files/352501", "https://ndownloader.figshare.com/files/352539", "https://ndownloader.figshare.com/files/352577", "https://ndownloader.figshare.com/files/352611", "https://ndownloader.figshare.com/files/352635"], "description"=>"<div><p>Mutations in the key enzyme of sialic acid biosynthesis, UDP-<em>N</em>-acetylglucosamine 2-epimerase/<em>N</em>-acetyl-mannosamine kinase, result in distal myopathy with rimmed vacuoles (DMRV)/hereditary inclusion body myopathy (HIBM) in humans. Sialic acid is an acidic monosaccharide that modifies non-reducing terminal carbohydrate chains on glycoproteins and glycolipids, and it plays an important role in cellular adhesions and interactions. In this study, we generated mice with a V572L point mutation in the GNE kinase domain. Unexpectedly, these mutant mice had no apparent myopathies or motor dysfunctions. However, they had a short lifespan and exhibited renal impairment with massive albuminuria. Histological analysis showed enlarged glomeruli with mesangial matrix deposition, leading to glomerulosclerosis and abnormal podocyte foot process morphologies in the kidneys. Glycan analysis using several lectins revealed glomerular epithelial cell hyposialylation, particularly the hyposialylation of podocalyxin, which is one of important molecules for the glomerular filtration barrier. Administering Neu5Ac to the mutant mice from embryonic stages significantly suppressed the albuminuria and renal pathology, and partially recovered the glomerular glycoprotein sialylation. These findings suggest that the nephrotic-like syndrome observed in these mutant mice resulted from impaired glomerular filtration due to the hyposialylation of podocyte glycoproteins, including podocalyxin. Furthermore, it was possible to prevent the nephrotic-like disease in these mice by beginning Neu5Ac treatment during gestation.</p> </div>", "links"=>[], "tags"=>["glycoprotein", "hyposialylation", "gives", "nephrotic-like", "prevented", "sialic", "gne", "v572l", "point-mutant", "mice"], "article_id"=>129655, "categories"=>["Biotechnology", "Biochemistry", "Marine Biology"], "users"=>["Mitutoshi Ito", "Kazushi Sugihara", "Tomoya Asaka", "Tadashi Toyama", "Toru Yoshihara", "Kengo Furuichi", "Takashi Wada", "Masahide Asano"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0029873.s001", "https://dx.doi.org/10.1371/journal.pone.0029873.s002", "https://dx.doi.org/10.1371/journal.pone.0029873.s003", "https://dx.doi.org/10.1371/journal.pone.0029873.s004", "https://dx.doi.org/10.1371/journal.pone.0029873.s005", "https://dx.doi.org/10.1371/journal.pone.0029873.s006", "https://dx.doi.org/10.1371/journal.pone.0029873.s007"], "stats"=>{"downloads"=>0, "page_views"=>36, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Glycoprotein_Hyposialylation_Gives_Rise_to_a_Nephrotic_Like_Syndrome_That_Is_Prevented_by_Sialic_Acid_Administration_in_GNE_V572L_Point_Mutant_Mice/129655", "title"=>"Glycoprotein Hyposialylation Gives Rise to a Nephrotic-Like Syndrome That Is Prevented by Sialic Acid Administration in GNE V572L Point-Mutant Mice", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2012-01-13 02:40:55"}
  • {"files"=>["https://ndownloader.figshare.com/files/692806"], "description"=>"<p>(A) Kidney sections stained with PNA, RCA-I, LTA, and DBA lectins (from left panels to right panels, respectively) in 3-month-old ht (upper panels) and mt (lower panels) mice. Scale bars: 120 µm. (B) Confocal laser scanning microscopic analysis of kidneys double-stained for PC and PNA, from 2-month-old ht (upper panels) and mt (lower panels) mice. Sections were stained with an anti-PC antibody (left panels; red), PNA (middle panels; green), and both (right panels; merge). Scale bars: 50 µm. (C) Confocal laser scanning microscopic analysis of kidneys double-stained for PC and PNA, from ht (upper panels) and mt (lower panels) mice at 8 days after birth. Sections were stained with the anti-PC antibody (left panels), PNA (middle panels; green), and both (right panels; merge). Scale bars: 50 µm.</p>", "links"=>[], "tags"=>["staining", "glycan", "podocalyxin"], "article_id"=>363252, "categories"=>["Biotechnology", "Biochemistry", "Marine Biology"], "users"=>["Mitutoshi Ito", "Kazushi Sugihara", "Tomoya Asaka", "Tadashi Toyama", "Toru Yoshihara", "Kengo Furuichi", "Takashi Wada", "Masahide Asano"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0029873.g005", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Lectin_staining_and_glycan_analysis_of_podocalyxin_PC_in_kidneys_/363252", "title"=>"Lectin staining and glycan analysis of podocalyxin (PC) in kidneys.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-13 00:54:12"}
  • {"files"=>["https://ndownloader.figshare.com/files/692932"], "description"=>"<p>(A) Western blotting analysis of kidney lysates, using an anti-PC antibody. PC bands in wt and ht mice were about 140–150 kDa (arrowheads), while those in mt mice showed a shift to higher molecular weights (bracketed). (B) Neuraminidase treatment shifted the PC bands in wt mice to higher molecular weights similar to those seen in untreated mt mice. (C to E) Glycan analysis of PC by lectin blotting. After immunoprecipitation with an anti-PC antibody, PC bands were detected by PC (C), RCA-I (D), and PNA (E).</p>", "links"=>[], "tags"=>["blot", "lectin", "pc"], "article_id"=>363376, "categories"=>["Biotechnology", "Biochemistry", "Marine Biology"], "users"=>["Mitutoshi Ito", "Kazushi Sugihara", "Tomoya Asaka", "Tadashi Toyama", "Toru Yoshihara", "Kengo Furuichi", "Takashi Wada", "Masahide Asano"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0029873.g006", "stats"=>{"downloads"=>3, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Western_blot_and_lectin_blot_PC_analysis_/363376", "title"=>"Western blot and lectin blot PC analysis.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-13 00:56:16"}
  • {"files"=>["https://ndownloader.figshare.com/files/692572"], "description"=>"<p>(A) PAS-stained kidney sections at 8 days, 3 months, 6 months and 12 months of age in ht (upper panels) and mt (lower panels) mice. Scale bars: 20 µm. (B) Electron microscopy of the glomeruli of wt (left panel) and mt mice (right panel) at 4 months old. Asterisks indicate flattened and fused podocyte foot processes.</p>", "links"=>[], "tags"=>["kidney"], "article_id"=>363018, "categories"=>["Biotechnology", "Biochemistry", "Marine Biology"], "users"=>["Mitutoshi Ito", "Kazushi Sugihara", "Tomoya Asaka", "Tadashi Toyama", "Toru Yoshihara", "Kengo Furuichi", "Takashi Wada", "Masahide Asano"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0029873.g003", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Histological_kidney_analysis_/363018", "title"=>"Histological kidney analysis.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-13 00:50:18"}
  • {"files"=>["https://ndownloader.figshare.com/files/692488"], "description"=>"<p>(<b>A</b>) Serum albumin in wt (open diamonds), ht (gray squares), and mt (black triangles) mice at 3 months (n = 4 wt, 2 ht, 6 mt), 6 months (n = 9 wt, 3 ht, 9 mt), and 12–15 months (n = 1 wt, 11 ht, 12 mt). Data of the wt and ht mice are presented in the same column. (<b>B</b>) Urinary albumin/creatinine ratio in wt (open diamonds), ht (gray squares), and mt (black triangles) mice at 3 months (n = 8 wt, 3 ht, 9 mt), 6 months (n = 4 wt, 5 ht, 11 mt), and 12–15 months (n = 5 ht, 5 mt). (<b>C</b>) Serum cystatin C at 3 months (n = 4 wt, 2 ht, 6 mt), 6 months (n = 6 wt, 1 ht, 6 mt), and 12–15 months (n = 1 wt, 5 ht, 6 mt). (<b>D</b>) SDS-PAGE analysis of urinary proteins in ht and mt mice at 10 days after birth; 8 µl of urine was separated by SDS-PAGE and stained by CBB. Arrowhead: albumin band (60 kDa). Results are shown as means (bars) with individual data points (A, B, C). *<i>P</i><0.05, **<i>P</i><0.01 (Mann-Whitney U-test).</p>", "links"=>[], "tags"=>["Nephrology", "biotechnology", "Biochemistry"], "article_id"=>362935, "categories"=>["Biotechnology", "Biochemistry", "Marine Biology"], "users"=>["Mitutoshi Ito", "Kazushi Sugihara", "Tomoya Asaka", "Tadashi Toyama", "Toru Yoshihara", "Kengo Furuichi", "Takashi Wada", "Masahide Asano"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0029873.g002", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Clinical_chemistry_tests_/362935", "title"=>"Clinical chemistry tests.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-13 00:48:55"}
  • {"files"=>["https://ndownloader.figshare.com/files/692398"], "description"=>"<p>(<b>A</b>) The sialic acid biosynthesis pathway. GNE has dual-function enzymatic activity, UDP-<i>N</i>-acetylglucosamine-2-epimerase (*1) and <i>N</i>-acetylmannosamine kinase (*2), in the cytosol. (<b>B</b>) Targeting strategy to create a point mutation in exon 10 of the mouse <i>GNE</i> gene: to make the targeted allele, the wild-type GNE locus was replaced with a targeting vector, which contained a point mutation in exon 10, a neo cassette and a DT-A cassette, by homologous recombination. The neo cassette, flanked by two loxP sites, was deleted by a Cre-expressing adenoviral vector to make the point mutation allele. Neo, neomycin-resistance gene; DT-A, diphtheria toxin A fragment; triangles, loxP sites; open boxes with numbers, exons; closed boxes, exon 10 containing the point mutation. (<b>C</b>) A base exchange from G (wild-type; wt) to C (mutant; mt) at the 1714 site was confirmed by DNA sequencing. (<b>D</b>) Quantitative RT-PCR analysis of the <i>GNE</i> mRNA level normalized to the <i>GAPDH</i> mRNA level in the quadriceps femoris and kidney of the mutant (mt), heterozygous (ht), and wild-type (wt) mice. (<b>E</b>) Survival ratio of mt (squares, n = 37), ht (triangles, n = 33), and wt mice (circles, n = 12), analyzed by Kaplan Meier methods.</p>", "links"=>[], "tags"=>["sialic", "biosynthesis", "pathway", "gne", "v572l", "point-mutant"], "article_id"=>362840, "categories"=>["Biotechnology", "Biochemistry", "Marine Biology"], "users"=>["Mitutoshi Ito", "Kazushi Sugihara", "Tomoya Asaka", "Tadashi Toyama", "Toru Yoshihara", "Kengo Furuichi", "Takashi Wada", "Masahide Asano"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0029873.g001", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_sialic_acid_biosynthesis_pathway_and_the_generation_of_GNE_V572L_point_mutant_mice_/362840", "title"=>"The sialic acid biosynthesis pathway and the generation of GNE V572L point-mutant mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-13 00:47:20"}
  • {"files"=>["https://ndownloader.figshare.com/files/693015"], "description"=>"<p>(A) Urinary albumin/creatinine ratios in untreated ht (green diamonds, n = 10) and mt (red triangles, n = 7) mice, and in Neu5Ac-treated ht (blue circles, n = 13) and mt (yellow squares, n = 8) mice at 1 and 2 months of age. ***<i>P</i><0.001 (Two-way ANOVA). (B) PAS-stained kidney sections of untreated ht (left panel) and mt (middle panel) mice, and of Neu5Ac-treated mt (right panel) mice at 2 months old. Scale bars: 20 µm. (C) Electron microscopy of the glomeruli of mt mice at 2 months old that were untreated (left panels) or treated (right panels) with Neu5Ac. Low (upper panels, scale bars: 1 µm) and high magnification pictures (lower panels, scale bars: 500 nm).</p>", "links"=>[], "tags"=>["histological", "mt", "mice", "treated"], "article_id"=>363461, "categories"=>["Biotechnology", "Biochemistry", "Marine Biology"], "users"=>["Mitutoshi Ito", "Kazushi Sugihara", "Tomoya Asaka", "Tadashi Toyama", "Toru Yoshihara", "Kengo Furuichi", "Takashi Wada", "Masahide Asano"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0029873.g007", "stats"=>{"downloads"=>1, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Urinary_and_histological_analysis_of_mt_mice_treated_with_Neu5Ac_/363461", "title"=>"Urinary and histological analysis of mt mice treated with Neu5Ac.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-13 00:57:41"}
  • {"files"=>["https://ndownloader.figshare.com/files/693146"], "description"=>"<p>(A) Kidneys double-stained for PC and PNA, from untreated ht (upper panels) and mt (middle panels) mice, and Neu5Ac-treated mt mice (bottom panels). Sections were stained with an anti-PC antibody (left panels; red), PNA (middle panels; green), and both (right panels; merge). Scale bars: 200 µm. (B) Ratio of PNA-positive to PC-positive glomeruli in the kidneys of 2-month-old untreated ht (green diamonds, n = 4) and mt (red triangles, n = 6) mice, and of Neu5Ac-treated ht (blue circles, n = 5) and mt (yellow squares, n = 8) mice. ***<i>P</i><0.001 (Two-way ANOVA).</p>", "links"=>[], "tags"=>["pna-positive", "glomeruli", "ratios", "neu5ac-treated", "mt"], "article_id"=>363588, "categories"=>["Biotechnology", "Biochemistry", "Marine Biology"], "users"=>["Mitutoshi Ito", "Kazushi Sugihara", "Tomoya Asaka", "Tadashi Toyama", "Toru Yoshihara", "Kengo Furuichi", "Takashi Wada", "Masahide Asano"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0029873.g008", "stats"=>{"downloads"=>0, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Reduced_PNA_positive_glomeruli_ratios_in_Neu5Ac_treated_mt_mice_/363588", "title"=>"Reduced PNA-positive glomeruli ratios in Neu5Ac-treated mt mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-13 00:59:48"}

PMC Usage Stats | Further Information

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Relative Metric

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