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Microtubule-associated protein tau is required for axonal neurite elaboration by neuroblastoma cells.

Identifieur interne : 004721 ( Main/Exploration ); précédent : 004720; suivant : 004722

Microtubule-associated protein tau is required for axonal neurite elaboration by neuroblastoma cells.

Auteurs : T B Shea [États-Unis] ; M L Beermann ; R A Nixon ; I. Fischer

Source :

RBID : pubmed:1433385

Descripteurs français

English descriptors

Abstract

NB2a/d1 neuroblastoma cells constitutively express multiple isoforms of the microtubule-associated protein tau and incorporate this protein into the axonal neurites elaborated during serum deprivation. To examine whether or not tau played an essential role in axonal outgrowth, cells cultured in serum-free medium were treated at 24 h intervals with antisense- and sense-oriented cDNA oligonucleotides (25 or 36 mers that span or are upstream of tau initiation codon) and were simultaneously serum deprived. Oligonucleotide uptake was confirmed by determination of intracellular levels of radiolabeled oligonucleotides. Treatment for 48 h with tau antisense oligonucleotides reversibly inhibited the expression of tau and the number of neurite-bearing cells compared with treatment with sense oligonucleotides. By contrast, tubulin expression was not affected. When cells were treated with antisense oligonucleotide simultaneously with serum deprivation, the initial outgrowth of neurites was unaffected, but continued neurite elongation was prevented. By contrast, neurite outgrowth at 4 h was inhibited when cells were pretreated with tau antisense 24 h before serum deprivation. Furthermore, intracellular delivery of anti-tau antiserum prevented neurite outgrowth and, in cells that had previously been deprived of serum for 24 h, induced retraction of existing neurites. These findings indicate that both the initiation and the continued outgrowth of neurites are dependent on tau and that pre-existing cytoplasmic pools of tau can mediate initial neuritogenesis.

DOI: 10.1002/jnr.490320308
PubMed: 1433385


Affiliations:


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Le document en format XML

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<nlm:affiliation>Laboratory for Cellular and Developmental Neurobiology, McLean Hospital, Belmont, Massachusetts.</nlm:affiliation>
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<name sortKey="Beermann, M L" sort="Beermann, M L" uniqKey="Beermann M" first="M L" last="Beermann">M L Beermann</name>
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<term>Axons (immunology)</term>
<term>Axons (physiology)</term>
<term>Base Sequence</term>
<term>DNA, Antisense (biosynthesis)</term>
<term>DNA, Antisense (immunology)</term>
<term>Electrophoresis, Polyacrylamide Gel</term>
<term>Immunoblotting</term>
<term>Immunohistochemistry</term>
<term>Molecular Sequence Data</term>
<term>Neurites (immunology)</term>
<term>Neurites (physiology)</term>
<term>Neuroblastoma (physiopathology)</term>
<term>Oligonucleotides (biosynthesis)</term>
<term>Precipitin Tests</term>
<term>Tumor Cells, Cultured</term>
<term>tau Proteins (immunology)</term>
<term>tau Proteins (physiology)</term>
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<term>ADN antisens (biosynthèse)</term>
<term>ADN antisens (immunologie)</term>
<term>Axones (immunologie)</term>
<term>Axones (physiologie)</term>
<term>Cellules cancéreuses en culture</term>
<term>Données de séquences moléculaires</term>
<term>Immunohistochimie</term>
<term>Immunotransfert</term>
<term>Neurites (immunologie)</term>
<term>Neurites (physiologie)</term>
<term>Neuroblastome (physiopathologie)</term>
<term>Oligonucléotides (biosynthèse)</term>
<term>Protéines tau (immunologie)</term>
<term>Protéines tau (physiologie)</term>
<term>Séquence nucléotidique</term>
<term>Tests aux précipitines</term>
<term>Électrophorèse sur gel de polyacrylamide</term>
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<term>ADN antisens</term>
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<term>Neurites</term>
<term>Protéines tau</term>
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<term>Axons</term>
<term>DNA, Antisense</term>
<term>Neurites</term>
<term>tau Proteins</term>
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<term>Axones</term>
<term>Neurites</term>
<term>Protéines tau</term>
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<term>Axons</term>
<term>Neurites</term>
<term>tau Proteins</term>
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<term>Neuroblastoma</term>
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<term>Electrophoresis, Polyacrylamide Gel</term>
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<term>Données de séquences moléculaires</term>
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<term>Immunotransfert</term>
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<div type="abstract" xml:lang="en">NB2a/d1 neuroblastoma cells constitutively express multiple isoforms of the microtubule-associated protein tau and incorporate this protein into the axonal neurites elaborated during serum deprivation. To examine whether or not tau played an essential role in axonal outgrowth, cells cultured in serum-free medium were treated at 24 h intervals with antisense- and sense-oriented cDNA oligonucleotides (25 or 36 mers that span or are upstream of tau initiation codon) and were simultaneously serum deprived. Oligonucleotide uptake was confirmed by determination of intracellular levels of radiolabeled oligonucleotides. Treatment for 48 h with tau antisense oligonucleotides reversibly inhibited the expression of tau and the number of neurite-bearing cells compared with treatment with sense oligonucleotides. By contrast, tubulin expression was not affected. When cells were treated with antisense oligonucleotide simultaneously with serum deprivation, the initial outgrowth of neurites was unaffected, but continued neurite elongation was prevented. By contrast, neurite outgrowth at 4 h was inhibited when cells were pretreated with tau antisense 24 h before serum deprivation. Furthermore, intracellular delivery of anti-tau antiserum prevented neurite outgrowth and, in cells that had previously been deprived of serum for 24 h, induced retraction of existing neurites. These findings indicate that both the initiation and the continued outgrowth of neurites are dependent on tau and that pre-existing cytoplasmic pools of tau can mediate initial neuritogenesis.</div>
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