KOPS - The Institutional Repository of the University of Konstanz

Time and space-resolved quantification of plasma membrane sialylation for measurements of cell function and neurotoxicity

Time and space-resolved quantification of plasma membrane sialylation for measurements of cell function and neurotoxicity

Cite This

Files in this item

Checksum: MD5:0f2978a1b000a28c35c81b37f04986b2

KRANASTER, Petra, Christiaan KARREMAN, Jeremias E.G.A. DOLD, Alice KREBS, Melina FUNKE, Anna-Katharina HOLZER, Stefanie KLIMA, Johanna NYFFELER, Stefan HELFRICH, Valentin WITTMANN, Marcel LEIST, 2020. Time and space-resolved quantification of plasma membrane sialylation for measurements of cell function and neurotoxicity. In: Archives of Toxicology. Springer Science and Business Media. 94(2), pp. 449-467. ISSN 0340-5761. eISSN 1432-0738. Available under: doi: 10.1007/s00204-019-02642-z

@article{Kranaster2020-02space-48086, title={Time and space-resolved quantification of plasma membrane sialylation for measurements of cell function and neurotoxicity}, year={2020}, doi={10.1007/s00204-019-02642-z}, number={2}, volume={94}, issn={0340-5761}, journal={Archives of Toxicology}, pages={449--467}, author={Kranaster, Petra and Karreman, Christiaan and Dold, Jeremias E.G.A. and Krebs, Alice and Funke, Melina and Holzer, Anna-Katharina and Klima, Stefanie and Nyffeler, Johanna and Helfrich, Stefan and Wittmann, Valentin and Leist, Marcel} }

<rdf:RDF xmlns:dcterms="http://purl.org/dc/terms/" xmlns:dc="http://purl.org/dc/elements/1.1/" xmlns:rdf="http://www.w3.org/1999/02/22-rdf-syntax-ns#" xmlns:bibo="http://purl.org/ontology/bibo/" xmlns:dspace="http://digital-repositories.org/ontologies/dspace/0.1.0#" xmlns:foaf="http://xmlns.com/foaf/0.1/" xmlns:void="http://rdfs.org/ns/void#" xmlns:xsd="http://www.w3.org/2001/XMLSchema#" > <rdf:Description rdf:about="https://kops.uni-konstanz.de/rdf/resource/123456789/48086"> <dc:creator>Funke, Melina</dc:creator> <dcterms:available rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2019-12-18T08:11:20Z</dcterms:available> <dcterms:isPartOf rdf:resource="https://kops.uni-konstanz.de/rdf/resource/123456789/29"/> <dc:creator>Holzer, Anna-Katharina</dc:creator> <dc:contributor>Krebs, Alice</dc:contributor> <dc:creator>Klima, Stefanie</dc:creator> <foaf:homepage rdf:resource="http://localhost:8080/jspui"/> <dc:creator>Kranaster, Petra</dc:creator> <dspace:hasBitstream rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/48086/1/Kranaster_2-18vj6nw36y4cq2.pdf"/> <dc:creator>Helfrich, Stefan</dc:creator> <dc:contributor>Holzer, Anna-Katharina</dc:contributor> <dc:contributor>Karreman, Christiaan</dc:contributor> <dc:contributor>Helfrich, Stefan</dc:contributor> <dc:contributor>Nyffeler, Johanna</dc:contributor> <dcterms:hasPart rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/48086/1/Kranaster_2-18vj6nw36y4cq2.pdf"/> <dc:contributor>Dold, Jeremias E.G.A.</dc:contributor> <dcterms:abstract xml:lang="eng">While there are many methods to quantify the synthesis, localization, and pool sizes of proteins and DNA during physiological responses and toxicological stress, only few approaches allow following the fate of carbohydrates. One of them is metabolic glycoengineering (MGE), which makes use of chemically modified sugars (CMS) that enter the cellular biosynthesis pathways leading to glycoproteins and glycolipids. The CMS can subsequently be coupled (via bio-orthogonal chemical reactions) to tags that are quantifiable by microscopic imaging. We asked here, whether MGE can be used in a quantitative and time-resolved way to study neuronal glycoprotein synthesis and its impairment. We focused on the detection of sialic acid (Sia), by feeding human neurons the biosynthetic precursor N-acetyl-mannosamine, modified by an azide tag. Using this system, we identified non-toxic conditions that allowed live cell labeling with high spatial and temporal resolution, as well as the quantification of cell surface Sia. Using combinations of immunostaining, chromatography, and western blotting, we quantified the percentage of cellular label incorporation and effects on glycoproteins such as polysialylated neural cell adhesion molecule. A specific imaging algorithm was used to quantify Sia incorporation into neuronal projections, as potential measure of complex cell function in toxicological studies. When various toxicants were studied, we identified a subgroup (mitochondrial respiration inhibitors) that affected neurite glycan levels several hours before any other viability parameter was affected. The MGE-based neurotoxicity assay, thus allowed the identification of subtle impairments of neurochemical function with very high sensitivity.</dcterms:abstract> <dc:creator>Krebs, Alice</dc:creator> <dcterms:title>Time and space-resolved quantification of plasma membrane sialylation for measurements of cell function and neurotoxicity</dcterms:title> <dc:creator>Nyffeler, Johanna</dc:creator> <dc:creator>Karreman, Christiaan</dc:creator> <dc:contributor>Wittmann, Valentin</dc:contributor> <dc:contributor>Funke, Melina</dc:contributor> <dcterms:rights rdf:resource="https://rightsstatements.org/page/InC/1.0/"/> <dc:contributor>Klima, Stefanie</dc:contributor> <dc:creator>Leist, Marcel</dc:creator> <dspace:isPartOfCollection rdf:resource="https://kops.uni-konstanz.de/rdf/resource/123456789/29"/> <bibo:uri rdf:resource="https://kops.uni-konstanz.de/handle/123456789/48086"/> <dc:creator>Wittmann, Valentin</dc:creator> <dspace:isPartOfCollection rdf:resource="https://kops.uni-konstanz.de/rdf/resource/123456789/28"/> <dcterms:isPartOf rdf:resource="https://kops.uni-konstanz.de/rdf/resource/123456789/28"/> <dc:contributor>Leist, Marcel</dc:contributor> <dc:date rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2019-12-18T08:11:20Z</dc:date> <dc:language>eng</dc:language> <void:sparqlEndpoint rdf:resource="http://localhost/fuseki/dspace/sparql"/> <dc:creator>Dold, Jeremias E.G.A.</dc:creator> <dcterms:issued>2020-02</dcterms:issued> <dc:contributor>Kranaster, Petra</dc:contributor> <dc:rights>terms-of-use</dc:rights> </rdf:Description> </rdf:RDF>

Downloads since Dec 18, 2019 (Information about access statistics)

Kranaster_2-18vj6nw36y4cq2.pdf 12

This item appears in the following Collection(s)

Search KOPS


Browse

My Account