Neural and Synaptic Defects in slytherin, a Zebrafish Model for Human Congenital Disorders of Glycosylation
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{"title"=>"Neural and synaptic defects in slytherin, a zebrafish model for human congenital disorders of glycosylation", "type"=>"journal", "authors"=>[{"first_name"=>"Yuanquan", "last_name"=>"Song", "scopus_author_id"=>"12242614100"}, {"first_name"=>"Jason R.", "last_name"=>"Willer", "scopus_author_id"=>"24172922300"}, {"first_name"=>"Paul C.", "last_name"=>"Scherer", "scopus_author_id"=>"35749159900"}, {"first_name"=>"Jessica A.", "last_name"=>"Panzer", "scopus_author_id"=>"7004099499"}, {"first_name"=>"Amy", "last_name"=>"Kugath", "scopus_author_id"=>"36621079600"}, {"first_name"=>"Emmanuel", "last_name"=>"Skordalakes", "scopus_author_id"=>"6603312926"}, {"first_name"=>"Ronald G.", "last_name"=>"Gregg", "scopus_author_id"=>"7102996154"}, {"first_name"=>"Gregory B.", "last_name"=>"Willer", "scopus_author_id"=>"8924630800"}, {"first_name"=>"Rita J.", "last_name"=>"Balice-Gordon", "scopus_author_id"=>"57189461307"}], "year"=>2010, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "pmid"=>"21060795", "pui"=>"359931389", "scopus"=>"2-s2.0-78149441426", "doi"=>"10.1371/journal.pone.0013743", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "sgr"=>"78149441426"}, "id"=>"7ed95149-dc3e-3b6b-ad4f-870c2128a815", "abstract"=>"Congenital disorder of glycosylation type IIc (CDG IIc) is characterized by mental retardation, slowed growth and severe immunodeficiency, attributed to the lack of fucosylated glycoproteins. While impaired Notch signaling has been implicated in some aspects of CDG IIc pathogenesis, the molecular and cellular mechanisms remain poorly understood. We have identified a zebrafish mutant slytherin (srn), which harbors a missense point mutation in GDP-mannose 4,6 dehydratase (GMDS), the rate-limiting enzyme in protein fucosylation, including that of Notch. Here we report that some of the mechanisms underlying the neural phenotypes in srn and in CGD IIc are Notch-dependent, while others are Notch-independent. We show, for the first time in a vertebrate in vivo, that defects in protein fucosylation leads to defects in neuronal differentiation, maintenance, axon branching, and synapse formation. Srn is thus a useful and important vertebrate model for human CDG IIc that has provided new insights into the neural phenotypes that are hallmarks of the human disorder and has also highlighted the role of protein fucosylation in neural development.", "link"=>"http://www.mendeley.com/research/neural-synaptic-defects-slytherin-zebrafish-model-human-congenital-disorders-glycosylation", "reader_count"=>16, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>2, "Researcher"=>7, "Student > Ph. D. Student"=>4, "Student > Postgraduate"=>1, "Student > Master"=>1, "Student > Bachelor"=>1}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>2, "Researcher"=>7, "Student > Ph. D. Student"=>4, "Student > Postgraduate"=>1, "Student > Master"=>1, "Student > Bachelor"=>1}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>3, "Agricultural and Biological Sciences"=>8, "Philosophy"=>1, "Pharmacology, Toxicology and Pharmaceutical Science"=>1, "Chemistry"=>2, "Social Sciences"=>1}, "reader_count_by_subdiscipline"=>{"Chemistry"=>{"Chemistry"=>2}, "Social Sciences"=>{"Social Sciences"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>8}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}, "Philosophy"=>{"Philosophy"=>1}}, "group_count"=>0}

Scopus | Further Information

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  • {"files"=>["https://ndownloader.figshare.com/files/821175"], "description"=>"<p><b>A</b>, <b>B.</b> Secondary motor neuron cell body number and patterning assayed with Zn5 immunostaining (18 embryos/3 carrier pairs for each). <b>B.</b> Higher magnification of boxed region in <b>A.</b> At 48–72 hpf, Zn5+ cell number is similar in <i>srn</i> and WT (<a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0013743#pone.0013743.s004\" target=\"_blank\">Fig. S4</a>), but the patterning of these cells is aberrant in <i>srn</i> embryos. Zn5+ cells are clumped in <i>srn</i> mutants (dashed blue bracket) compared to WT embryos (solid blue bracket). <i>dla</i> mutants do not have defects in Zn5+ cell number (<a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0013743#pone.0013743.s004\" target=\"_blank\">Fig. S4</a>), but have aberrant Zn5+ cell patterning as in <i>srn</i> mutants (dashed blue bracket). <i>mib</i> mutants and high dose DAPT treated embryos have aberrant Zn5+ cell number (<a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0013743#pone.0013743.s004\" target=\"_blank\">Fig. S4</a>) and patterning (dashed blue bracket). Medium dose DAPT treated embryos show aberrant Zn5+ cell patterning defects (dashed blue bracket), without an obvious change in cell number (<a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0013743#pone.0013743.s004\" target=\"_blank\">Fig. S4</a>), as in <i>srn</i>. The dorsal projecting nerve is absent in <i>srn</i> mutants (dashed pink oval) compared to WT (solid pink oval), consistent with increased cell death; this nerve is present in <i>dla</i> and <i>des</i> mutants (solid pink oval); <i>des</i> also has other motor axon pathfinding errors. In <i>mib</i> mutants and high and medium dose DAPT treated embryos, the dorsal projecting nerve is absent (dashed pink oval). <b>C.</b> In WT embryos at 48 hpf, two Mauthner neurons are present (dorsal view of hindbrain). In <i>srn</i>, <i>des</i>, <i>dla</i>, <i>mib</i> and high dose DAPT treated embryos, Mauthner neuron number is increased (dashed red brackets), with the largest increase observed in <i>mib</i> (12 embryos, 3 carrier pairs for each). <b>D.</b> In the spinal cord, the number of GFAP+ glial cells is reduced in <i>srn</i> and <i>dla</i> mutants and medium dose DAPT treated embryos compared to WT and <i>des</i> embryos at 48–72 hpf. In <i>mib</i> and high dose DAPT treated embryos, a more dramatic reduction is observed. The GFAP labeling that remains in <i>mib</i> mutants is likely to be in Rohon-Beard neurons dorsally and secondary motor neurons ventrally and is easily separated morphologically and based on its location from glial processes, thus does not interfere with analyses of glial defects (18 embryos, 3 carrier pairs for each). <b>E.</b> Overlay showing both the Zn5 and GFAP staining in the spinal cord. Scale bars  = 40 µm.</p>", "links"=>[], "tags"=>["notch-delta", "signaling", "accounts"], "article_id"=>491536, "categories"=>["Neuroscience", "Developmental Biology"], "users"=>["Yuanquan Song", "Jason R. Willer", "Paul C. Scherer", "Jessica A. Panzer", "Amy Kugath", "Emmanuel Skordalakes", "Ronald G. Gregg", "Gregory B. Willer", "Rita J. Balice-Gordon"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0013743.g004", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Reduction_in_Notch_Delta_signaling_accounts_for_some_srn_phenotypes_/491536", "title"=>"Reduction in Notch-Delta signaling accounts for some <i>srn</i> phenotypes.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-10-29 00:25:36"}
  • {"files"=>["https://ndownloader.figshare.com/files/821680"], "description"=>"<p><b>A.</b> qRT-PCR assessment of fold change in <i>hes5</i>, <i>her4</i> and <i>heyl</i> expression in WT, <i>srn</i> and <i>mib</i> mutant embryos at 48 hpf, normalized to <i>β-actin1</i>. <i>hes5</i>, <i>her4</i> and <i>heyl</i> expression is dramatically reduced in <i>srn</i>, similar to those in <i>mib</i>, but to a lesser extent. (3–5 experiments, 20 embryos each, one-way ANOVA, Bonferroni's Multiple Comparison Test, ** p<0.001, * p<0.5). <b>B. </b><i>hes5</i>, <i>her4</i> and <i>heyl in situ</i> hybridization at 48 hpf confirm reduced expression in the brain and spinal cord in <i>srn</i> mutants compared to WT (>30 embryos for each). Scale bar  = 100 µm.</p>", "links"=>[], "tags"=>["mutants", "showed", "aberrant", "notch", "responsive", "genes"], "article_id"=>492042, "categories"=>["Neuroscience", "Developmental Biology"], "users"=>["Yuanquan Song", "Jason R. Willer", "Paul C. Scherer", "Jessica A. Panzer", "Amy Kugath", "Emmanuel Skordalakes", "Ronald G. Gregg", "Gregory B. Willer", "Rita J. Balice-Gordon"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0013743.g007", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_srn_mutants_showed_aberrant_expression_of_Notch_responsive_genes_similar_to_mib_mutants_/492042", "title"=>"<i>srn</i> mutants showed aberrant expression of Notch responsive genes similar to <i>mib</i> mutants.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-10-29 00:34:02"}
  • {"files"=>["https://ndownloader.figshare.com/files/821834"], "description"=>"<p><b>A–X.</b> Presynaptic terminals (green) and postsynaptic AChR clusters (red) in 24 and 48 embryos from WT (A, G, M, S), <i>srn</i> (B, H, N, T), <i>des</i> (C, I, O, U), <i>dla</i> (D, J, P, V), <i>mib</i> (E, K, Q, W) and DAPT treated embryos (F, L, R, X). Boxed regions are shown at higher magnification at 24 (G–L) and 48 hpf (S–X; 3 hemisegments in each of 20 embryos, 3 carrier pairs for each). Scale bar  = 20 µm. <b>Y.</b> Presynaptic terminal, axon and synapse area at the choice point was significantly increased in all mutants, except in <i>mib</i> and DAPT treated embryos at 48 hpf, compared to WT (one-way ANOVA, Bonferroni's Multiple Comparison Test, ** p<0.01, *** p<0.001).</p>", "links"=>[], "tags"=>["mutants", "defects", "neuromuscular", "synaptogenesis", "notch-delta"], "article_id"=>492195, "categories"=>["Neuroscience", "Developmental Biology"], "users"=>["Yuanquan Song", "Jason R. Willer", "Paul C. Scherer", "Jessica A. Panzer", "Amy Kugath", "Emmanuel Skordalakes", "Ronald G. Gregg", "Gregory B. Willer", "Rita J. Balice-Gordon"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0013743.g008", "stats"=>{"downloads"=>1, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Slytherin_mutants_exhibit_defects_in_neuromuscular_synaptogenesis_due_in_part_to_reduction_in_Notch_Delta_signaling_/492195", "title"=>"<i>Slytherin</i> mutants exhibit defects in neuromuscular synaptogenesis due in part to reduction in Notch-Delta signaling.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-10-29 00:36:35"}
  • {"files"=>["https://ndownloader.figshare.com/files/822008"], "description"=>"<p><b>A.</b> In <i>srn</i> mutants at 72 hpf, the OPL (arrowheads) and IPL (arrows) are disorganized; this is not seen in <i>des</i> or <i>dla</i> mutants. In <i>mib</i> mutants, retinal ganglion and other cells die, resulting in decreased retina neuropil (rightmost panel; 8 embryos, 2 carrier pairs for each). Scale bar  = 20 µm. <b>B.</b> In <i>srn</i> mutants, retinal ganglion cell axons grow out to the optic chiasm and to optic tectum, but axon branches are aberrantly distributed within tectum (dashed white circle) and medial axon projections are shifted towards the midline (compare solid arrow and dashed arrow). Virtually all retina was dye labeled, and the labeling pattern was consistent across experiments, thus these defecs aren't due to incomplete dye uptake or labeling in <i>srn</i> mutants. These phenotypes are not present in <i>des</i> or <i>dla</i> mutants, and are also different from <i>mib</i> mutants, in which retinal ganglion cell axonal projections to optic tectum are dramatically reduced, as a consequence of retinal ganglion cell death. <i>Mib</i> mutants also displayed axon pathfinding errors at the optic chiasm; axons branched anterior to the optic chiasm (dashed square), while branching within tectum was dramatically reduced (dashed white circle; 15 embryos, 3 carrier pairs for each). Scale bar  = 20 µm. <b>C</b>, <b>D.</b> Topographic mapping of axon projections to optic tectum; dorsonasal (DN) and ventrotemporal (VT) axons were with DiI or DiD. DN and VT axon projections within tectum are aberrant in <i>srn</i> mutants, as is overlap dorsally (C) and laterally (D; 8 embryos, 2 carrier pairs for each). Scale bar  = 20 µm. <b>E.</b> In retina and optic tectum (white circle), the overall cellular lamination pattern as assessed by DAPI staining is grossly normal in <i>srn</i> mutants (compare bottom left panels, WT and <i>srn</i>). TUNEL staining showed that increased cell death was observed in the retina of <i>srn</i> mutants compared to WT embryos at 72 hpf; no difference in cell death in the optic tectum was observed in <i>srn</i> mutants compared to WT embryos at 72 hpf (color overlay, right most panels; 2–3 embryos, 1 carrier pair). Scale bar  = 20 µm.</p>", "links"=>[], "tags"=>["mutants", "defects", "axon", "branching", "cns", "synaptic", "connectivity", "notch-delta"], "article_id"=>492361, "categories"=>["Neuroscience", "Developmental Biology"], "users"=>["Yuanquan Song", "Jason R. Willer", "Paul C. Scherer", "Jessica A. Panzer", "Amy Kugath", "Emmanuel Skordalakes", "Ronald G. Gregg", "Gregory B. Willer", "Rita J. Balice-Gordon"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0013743.g009", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Slytherin_mutants_exhibit_defects_in_axon_branching_and_CNS_synaptic_connectivity_that_are_independent_of_Notch_Delta_signaling_/492361", "title"=>"<i>Slytherin</i> mutants exhibit defects in axon branching and CNS synaptic connectivity that are independent of Notch-Delta signaling.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-10-29 00:39:21"}
  • {"files"=>["https://ndownloader.figshare.com/files/820847"], "description"=>"<p><b>A.</b> AAL staining of WT embryos at 48 hpf showed that protein fucosylation is present throughout the embryo (10–15 embryos/2–3 adult pairs for all analyses). <b>B–E</b>. Protein fucosylation is prominent in several neural tissues including retina (lateral view), optic tectum (dorsal view), hindbrain (lateral view), spinal cord (lateral view) and neuromuscular synapses (lateral view of axial muscle). <b>F.</b> Protein fucosylation is dramatically reduced in <i>srn</i> mutants. Scale bar  = 20 µm. <b>G–J</b>. Reduced protein fucosylation in several neural tissues. Scale bar  = 20 µm. <b>K</b>. Protein fucosylation at neuromuscular synapses in WT embryos at 48 hpf, as shown by the colocalization of AAL staining (green) with markers for presynaptic axons and nerve terminals (SV2, red) and postsynaptic AChR clusters (α-bungarotoxin, blue). <b>L–N.</b> Higher magnification of boxed region in <b>K. </b><b>O.</b> Protein fucosylation is reduced at <i>srn</i> neuromuscular synapses. Scale bar  = 20 µm. <b>P–R</b>. Higher magnification of boxed region in <b>O.</b> Synapse area is significantly increased in <i>srn</i> mutants, e.g., at the choice point (compare dashed bracket in Q to solid bracket in M). Scale bar  = 20 µm.</p>", "links"=>[], "tags"=>["mutants", "reduced", "fucosylation", "aal"], "article_id"=>491210, "categories"=>["Neuroscience", "Developmental Biology"], "users"=>["Yuanquan Song", "Jason R. Willer", "Paul C. Scherer", "Jessica A. Panzer", "Amy Kugath", "Emmanuel Skordalakes", "Ronald G. Gregg", "Gregory B. Willer", "Rita J. Balice-Gordon"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0013743.g002", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_slytherin_mutants_exhibit_reduced_protein_fucosylation_as_measured_by_AAL_staining_/491210", "title"=>"<i>slytherin</i> mutants exhibit reduced protein fucosylation as measured by AAL staining.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-10-29 00:20:10"}
  • {"files"=>["https://ndownloader.figshare.com/files/821051"], "description"=>"<p><b>A.</b> External <i>srn</i> phenotypes including bent tail and aberrant hindbrain (bracket) are rescued by GDP-fucose supplementation (3 embryos). Scale bar  = 100 µm. <b>B.</b> GDP-fucose injection significantly reduced the percentage of mutants from 28.6±1.2% to 8.4±2.7% (576 embryos, 4 carrier pairs; Student's test, p<0.0001). <b>C.</b> After GDP-fucose supplementation (2 embryos), protein fucosylation as assessed by AAL staining at 48 hpf is rescued throughout <i>srn</i> embryos, to levels similar to those in WT embryos. Scale bar  = 100 µm.</p>", "links"=>[], "tags"=>["exogenous", "gdp-fucose", "rescues", "phenotypes", "restores", "aal"], "article_id"=>491404, "categories"=>["Neuroscience", "Developmental Biology"], "users"=>["Yuanquan Song", "Jason R. Willer", "Paul C. Scherer", "Jessica A. Panzer", "Amy Kugath", "Emmanuel Skordalakes", "Ronald G. Gregg", "Gregory B. Willer", "Rita J. Balice-Gordon"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0013743.g003", "stats"=>{"downloads"=>1, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Supplementation_with_exogenous_GDP_fucose_rescues_srn_external_phenotypes_and_restores_AAL_staining_/491404", "title"=>"Supplementation with exogenous GDP-fucose rescues <i>srn</i> external phenotypes and restores AAL staining.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-10-29 00:23:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/821374"], "description"=>"<p><b>A–C..... </b><i>mib</i> excludes <i>srn</i> phenotypes. <b>A.</b> AAL staining is reduced in <i>srn</i> and <i>srn</i> + <i>mib</i> double mutants, but not in <i>mib</i>. <b>B..... </b><i>srn</i> + <i>mib</i> double mutants showed reduction of secondary motor neurons, more severe than <i>srn</i> but similar to <i>mib</i> alone. <b>C..... </b><i>srn</i> + <i>mib</i> double mutants have reduced GFAP+ glia, more severe than <i>srn</i>, but similar to <i>mib</i> alone (15 embryos, 2 carrier pairs for each). Scale bar  = 40 µm. <b>D–F.</b> DAPT treatment excludes <i>srn</i> phenotypes. <b>D.</b> AAL staining is reduced in <i>srn</i> and <i>srn</i> mutants treated with DAPT, but not in DAPT treated embryos. <b>E..... </b><i>srn</i> mutants treated with DAPT showed reduction of secondary motor neurons, more severe than <i>srn</i> but similar to DAPT treated embryos. <b>F..... </b><i>srn</i> mutants treated with DAPT showed reduction of GFAP+ glia, more severe than <i>srn</i>, but similar to DAPT treated WT embryos (10 embryos, 2 carrier pairs for each). Scale bar  = 40 µm.</p>", "links"=>[], "tags"=>["dapt"], "article_id"=>491741, "categories"=>["Neuroscience", "Developmental Biology"], "users"=>["Yuanquan Song", "Jason R. Willer", "Paul C. Scherer", "Jessica A. Panzer", "Amy Kugath", "Emmanuel Skordalakes", "Ronald G. Gregg", "Gregory B. Willer", "Rita J. Balice-Gordon"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0013743.g005", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_mib_and_DAPT_treatment_exclude_srn_phenotypes_/491741", "title"=>"<i>mib</i> and DAPT treatment exclude <i>srn</i> phenotypes.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-10-29 00:29:01"}
  • {"files"=>["https://ndownloader.figshare.com/files/409495", "https://ndownloader.figshare.com/files/409524", "https://ndownloader.figshare.com/files/409551", "https://ndownloader.figshare.com/files/409585", "https://ndownloader.figshare.com/files/409604", "https://ndownloader.figshare.com/files/409652", "https://ndownloader.figshare.com/files/409692", "https://ndownloader.figshare.com/files/409726"], "description"=>"<div><p>Congenital disorder of glycosylation type IIc (CDG IIc) is characterized by mental retardation, slowed growth and severe immunodeficiency, attributed to the lack of fucosylated glycoproteins. While impaired Notch signaling has been implicated in some aspects of CDG IIc pathogenesis, the molecular and cellular mechanisms remain poorly understood. We have identified a zebrafish mutant <em>slytherin</em> (<em>srn</em>), which harbors a missense point mutation in GDP-mannose 4,6 dehydratase (GMDS), the rate-limiting enzyme in protein fucosylation, including that of Notch. Here we report that some of the mechanisms underlying the neural phenotypes in <em>srn</em> and in CGD IIc are Notch-dependent, while others are Notch-independent. We show, for the first time in a vertebrate <em>in vivo</em>, that defects in protein fucosylation leads to defects in neuronal differentiation, maintenance, axon branching, and synapse formation. <em>Srn</em> is thus a useful and important vertebrate model for human CDG IIc that has provided new insights into the neural phenotypes that are hallmarks of the human disorder and has also highlighted the role of protein fucosylation in neural development.</p></div>", "links"=>[], "tags"=>["neural", "synaptic", "defects", "zebrafish", "congenital", "disorders", "glycosylation"], "article_id"=>140880, "categories"=>["Neuroscience", "Developmental Biology"], "users"=>["Yuanquan Song", "Jason R. Willer", "Paul C. Scherer", "Jessica A. Panzer", "Amy Kugath", "Emmanuel Skordalakes", "Ronald G. Gregg", "Gregory B. Willer", "Rita J. Balice-Gordon"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0013743.s001", "https://dx.doi.org/10.1371/journal.pone.0013743.s002", "https://dx.doi.org/10.1371/journal.pone.0013743.s003", "https://dx.doi.org/10.1371/journal.pone.0013743.s004", "https://dx.doi.org/10.1371/journal.pone.0013743.s005", "https://dx.doi.org/10.1371/journal.pone.0013743.s006", "https://dx.doi.org/10.1371/journal.pone.0013743.s007", "https://dx.doi.org/10.1371/journal.pone.0013743.s008"], "stats"=>{"downloads"=>23, "page_views"=>18, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Neural_and_Synaptic_Defects_in_slytherin_a_Zebrafish_Model_for_Human_Congenital_Disorders_of_Glycosylation/140880", "title"=>"Neural and Synaptic Defects in <em>slytherin</em>, a Zebrafish Model for Human Congenital Disorders of Glycosylation", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2010-10-29 00:14:40"}
  • {"files"=>["https://ndownloader.figshare.com/files/821531"], "description"=>"<p><b>A.</b> AAL staining is reduced in <i>srn</i>, <i>gmds</i> morphants and <i>gmds</i> morphants overexpressing NICD, but not in WT embryos or WT embryos overexpressing NICD. <b>B.</b> WT and WT overexpressing NICD had normal Zn5+ cell patterning. <i>srn</i> and <i>gmds</i> MO showed Zn5+ cell patterning defects which was rescued by NICD overexpression in <i>gmds</i> morphants. <b>C.</b> WT and WT overexpressing NICD had normal GFAP+ glia cells in spinal cord. <i>srn</i> and <i>gmds</i> morphants had reduced GFAP+ glia cells, rescued by NICD overexpression in <i>gmds</i> morphants (>10 embryos in each experiment). Scale bar  = 40 µm. <b>D–E.</b> Quantification of Zn5+ cell patterning defects. There are 3–5 Zn5+ cells every 20 µm in WT, WT overexpressing NICD and <i>gmds</i> MO overexpressing NICD; compared to 1–8 in <i>srn</i> and <i>gmds</i> MO embryos. D, data from a representative embryo; E, distribution of all embryos (4–6 embryos; Kolmogorov-Smirnov test, * p<0.05).</p>", "links"=>[], "tags"=>["rescues", "neuro-", "gliogenesis"], "article_id"=>491889, "categories"=>["Neuroscience", "Developmental Biology"], "users"=>["Yuanquan Song", "Jason R. Willer", "Paul C. Scherer", "Jessica A. Panzer", "Amy Kugath", "Emmanuel Skordalakes", "Ronald G. Gregg", "Gregory B. Willer", "Rita J. Balice-Gordon"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0013743.g006", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_NICD_rescues_srn_neuro_and_gliogenesis_phenotypes_/491889", "title"=>"NICD rescues <i>srn</i> neuro- and gliogenesis phenotypes.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-10-29 00:31:29"}
  • {"files"=>["https://ndownloader.figshare.com/files/820688"], "description"=>"<p><b>A.</b> External <i>srn</i> phenotypes at 48 hpf include a bent tail (80% dorsal (534 embryos, 8 carrier pairs)) and aberrant hindbrain formation (brackets). Scale bar  = 100 µm. <b>B.</b> Genetic and physical map of the <i>srn</i> locus (red arrow), including SSLP markers, number of recombinants, BAC clones and megabase positions from Ensembl Zv7. <b>C</b>, <b>D.</b> In <i>srn</i>, <i>Gmds</i> mutation is G to T (C, red box) resulting in a Glycine to Valine conversion (D, red box). GMDS amino acid sequence is highly conserved. <b>E.</b> Schematic of <i>srn</i> mutation in the short-chain dehydrogenase/reductase (SDR) domain of GMDS. <b>F.</b> Two splice variants exist in <i>gmds</i> mRNA, with (<i>gmds-L</i>, 377 aa) or without (<i>gmds-S</i>, 370 aa) exon 4. <i>Gmds</i> alternative splicing is not altered in <i>srn</i> mutants. <b>G.</b> Injection of <i>gmds</i> mRNA rescues <i>srn</i> mutants. Compared to uninjected embryos, 28.6±1.2% of embryos were mutant when scored by external phenotypes (3413 embryos, 27 carrier pairs). In embryos injected with WT <i>gmds-gfp</i> mRNA, the percentage of mutants scored by external phenotypes was significantly decreased, to ca. 5% (<i>gmds-wtL-gfp</i> 5.4±2.5%, 401 embryos, 3 carrier pairs; <i>gmds-wtS-gfp</i> 5.1±0.6%, 587 embryos, 4 carrier pairs; one-way ANOVA, followed by Dunn's pairwise comparison, p<0.05). The percentage of embryos with mutant external phenotypes was unchanged in embryos injected with mutant <i>gmds-gfp</i> mRNA (<i>gmds-srnL-gfp</i> 30.2±0.9%, 387 embryos, 3 carrier pairs; <i>gmds-wtS-gfp</i> 25.1±1.9%, 516 embryos, 4 carrier pairs). This mRNA rescue experiment confirms that <i>gmds</i> is the gene responsible for <i>srn</i> mutation.</p>", "links"=>[], "tags"=>["cloning", "mrna"], "article_id"=>491050, "categories"=>["Neuroscience", "Developmental Biology"], "users"=>["Yuanquan Song", "Jason R. Willer", "Paul C. Scherer", "Jessica A. Panzer", "Amy Kugath", "Emmanuel Skordalakes", "Ronald G. Gregg", "Gregory B. Willer", "Rita J. Balice-Gordon"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0013743.g001", "stats"=>{"downloads"=>1, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Slytherin_external_phenotype_genotype_cloning_and_mRNA_rescue_of_srn_mutants_/491050", "title"=>"<i>Slytherin</i> external phenotype, genotype, cloning and mRNA rescue of <i>srn</i> mutants.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-10-29 00:17:30"}

PMC Usage Stats | Further Information

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Relative Metric

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