Ectopic expression of the TrkA receptor in adult dopaminergic mesencephalic neurons promotes retrograde axonal NGF transport and NGF-dependent neuroprotection
Identifieur interne : 000281 ( France/Analysis ); précédent : 000280; suivant : 000282Ectopic expression of the TrkA receptor in adult dopaminergic mesencephalic neurons promotes retrograde axonal NGF transport and NGF-dependent neuroprotection
Auteurs : Benoit Melchior [France] ; Véronique Nerriere-Daguin [France] ; David-A. Laplaud [France] ; Séverine Remy [France] ; Sandrine Wiertlewski [France] ; Isabelle Neveu [France] ; Philippe Naveilhan [France] ; Susan O. Meakin [Canada] ; Philippe Brachet [France]Source :
- Experimental neurology : (Print) [ 0014-4886 ] ; 2003.
Descripteurs français
- Pascal (Inist)
English descriptors
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Abstract
A recombinant adeno-associated virus (rAAV) was used to investigate the impact of an ectopic expression of the NGF high-affinity receptor in adult neurons. The rat TrkA cDNA cloned in a pCMX vector was first tagged with a human c-Myc sequence. The resulting vector was shown to encode a functional receptor which promoted the expression of TrkA immunoreactivity upon transfection of 293 fibroblasts or nnr5 cells, a TrkA-defective variant of PC 12 cells. These cells also accumulate TrkA transcripts upon transfection and extended neurites in the presence of NGF. Therefore, the TrkAmyc cassette was inserted into the pSSV9 plasmid. The new vectors shared properties similar to pCMX TrkAmyc in 293 and nnr5 cells and enabled the preparation of rAAV TrkAmyc viruses. Unilateral injection of this rAAV into the substantia nigra (SN) resulted in a protracted expression of TrkA (or c-Myc) immunoreactivity in numerous cell bodies, including tyrosine-hydroxylase (TH)-positive dopaminergic neurons. The presence of TrkA receptors in corresponding striatal dopaminergic endings was demonstrated by the advent of a striato-nigral retrograde axonal transport of 125I-NGF. Likewise, ectopic expression of TrkA in neurons of the parafascicular thalamic nucleus promoted a striatofuge transport of NGF toward this structure. To investigate whether ectopic expression of TrkA in SN neurons may confer neuroprotection, lesions were induced by 6-hydroxydopamine in striata located ipsilateral to the virus injection site. NGF or vehicle were next delivered dorsally to the virus-treated SN for 2 weeks, before sacrifice and processing of brains for TH-immunohistochemistry. NGF treatment, in contrast to treatment with vehicle, significantly enhanced the number of dopaminergic neurons counted in the lesioned SN. These data suggest that ectopic TrkA can mediate the trophic actions of NGF and influence neuronal plasticity in vivo.
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Laplaud</name><affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Institut National de la Samé et de la Recherche Médicale, Unité 437</s1><s2>Nantes</s2><s3>FRA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>France</country><placeName><region type="region">Pays de la Loire</region><region type="old region">Pays de la Loire</region><settlement type="city">Nantes</settlement></placeName></affiliation><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Clinique Neurologique, Centre Hospitalier Universitaire</s1><s2>44093 Nantes</s2><s3>FRA</s3><sZ>3 aut.</sZ><sZ>5 aut.</sZ></inist:fA14><country>France</country><wicri:noRegion>44093 Nantes</wicri:noRegion><wicri:noRegion>Centre Hospitalier Universitaire</wicri:noRegion><wicri:noRegion>44093 Nantes</wicri:noRegion></affiliation></author><author><name sortKey="Remy, Severine" sort="Remy, Severine" uniqKey="Remy S" first="Séverine" last="Remy">Séverine Remy</name><affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Institut National de la Samé et de la Recherche Médicale, Unité 437</s1><s2>Nantes</s2><s3>FRA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>France</country><placeName><region type="region">Pays de la Loire</region><region type="old region">Pays de la Loire</region><settlement type="city">Nantes</settlement></placeName></affiliation></author><author><name sortKey="Wiertlewski, Sandrine" sort="Wiertlewski, Sandrine" uniqKey="Wiertlewski S" first="Sandrine" last="Wiertlewski">Sandrine Wiertlewski</name><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Clinique Neurologique, Centre Hospitalier Universitaire</s1><s2>44093 Nantes</s2><s3>FRA</s3><sZ>3 aut.</sZ><sZ>5 aut.</sZ></inist:fA14><country>France</country><wicri:noRegion>44093 Nantes</wicri:noRegion><wicri:noRegion>Centre Hospitalier Universitaire</wicri:noRegion><wicri:noRegion>44093 Nantes</wicri:noRegion></affiliation></author><author><name sortKey="Neveu, Isabelle" sort="Neveu, Isabelle" uniqKey="Neveu I" first="Isabelle" last="Neveu">Isabelle Neveu</name><affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Institut National de la Samé et de la Recherche Médicale, Unité 437</s1><s2>Nantes</s2><s3>FRA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>France</country><placeName><region type="region">Pays de la Loire</region><region type="old region">Pays de la Loire</region><settlement type="city">Nantes</settlement></placeName></affiliation></author><author><name sortKey="Naveilhan, Philippe" sort="Naveilhan, Philippe" uniqKey="Naveilhan P" first="Philippe" last="Naveilhan">Philippe Naveilhan</name><affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Institut National de la Samé et de la Recherche Médicale, Unité 437</s1><s2>Nantes</s2><s3>FRA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>France</country><placeName><region type="region">Pays de la Loire</region><region type="old region">Pays de la Loire</region><settlement type="city">Nantes</settlement></placeName></affiliation></author><author><name sortKey="Meakin, Susan O" sort="Meakin, Susan O" uniqKey="Meakin S" first="Susan O." last="Meakin">Susan O. Meakin</name><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>The Robarts Research Institute</s1><s2>London, Ontario</s2><s3>CAN</s3><sZ>8 aut.</sZ></inist:fA14><country>Canada</country><wicri:noRegion>The Robarts Research Institute</wicri:noRegion></affiliation></author><author><name sortKey="Brachet, Philippe" sort="Brachet, Philippe" uniqKey="Brachet P" first="Philippe" last="Brachet">Philippe Brachet</name><affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Institut National de la Samé et de la Recherche Médicale, Unité 437</s1><s2>Nantes</s2><s3>FRA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>France</country><placeName><region type="region">Pays de la Loire</region><region type="old region">Pays de la Loire</region><settlement type="city">Nantes</settlement></placeName></affiliation></author></analytic><series><title level="j" type="main">Experimental neurology : (Print)</title><title level="j" type="abbreviated">Exp. neurol. : (Print)</title><idno type="ISSN">0014-4886</idno><imprint><date when="2003">2003</date></imprint></series></biblStruct></sourceDesc><seriesStmt><title level="j" type="main">Experimental neurology : (Print)</title><title level="j" type="abbreviated">Exp. neurol. : (Print)</title><idno type="ISSN">0014-4886</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Alzheimer disease</term><term>Aminoacid</term><term>Animal</term><term>C-Onc gene</term><term>Complementary DNA</term><term>Dopaminergic neuron</term><term>Fibroblast</term><term>Gene therapy</term><term>Immunohistochemistry</term><term>Nerve growth factor</term><term>Parkinson disease</term><term>Rat</term><term>Recombinant virus</term><term>Transfection</term><term>Tyrosine</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Fibroblaste</term><term>Tyrosine</term><term>Démence Alzheimer</term><term>Immunohistochimie</term><term>Thérapie génique</term><term>Parkinson maladie</term><term>DNA complémentaire</term><term>Neurone dopaminergique</term><term>Facteur croissance nerf</term><term>Virus recombinant</term><term>Animal</term><term>Rat</term><term>Gène onc cellulaire</term><term>Aminoacide</term><term>Transfection</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">A recombinant adeno-associated virus (rAAV) was used to investigate the impact of an ectopic expression of the NGF high-affinity receptor in adult neurons. The rat TrkA cDNA cloned in a pCMX vector was first tagged with a human c-Myc sequence. The resulting vector was shown to encode a functional receptor which promoted the expression of TrkA immunoreactivity upon transfection of 293 fibroblasts or nnr5 cells, a TrkA-defective variant of PC 12 cells. These cells also accumulate TrkA transcripts upon transfection and extended neurites in the presence of NGF. Therefore, the TrkA<sub>myc</sub> cassette was inserted into the pSSV9 plasmid. The new vectors shared properties similar to pCMX TrkA<sub>myc</sub> in 293 and nnr5 cells and enabled the preparation of rAAV TrkA<sub>myc</sub> viruses. Unilateral injection of this rAAV into the substantia nigra (SN) resulted in a protracted expression of TrkA (or c-Myc) immunoreactivity in numerous cell bodies, including tyrosine-hydroxylase (TH)-positive dopaminergic neurons. The presence of TrkA receptors in corresponding striatal dopaminergic endings was demonstrated by the advent of a striato-nigral retrograde axonal transport of <sup>125</sup>I-NGF. Likewise, ectopic expression of TrkA in neurons of the parafascicular thalamic nucleus promoted a striatofuge transport of NGF toward this structure. To investigate whether ectopic expression of TrkA in SN neurons may confer neuroprotection, lesions were induced by 6-hydroxydopamine in striata located ipsilateral to the virus injection site. NGF or vehicle were next delivered dorsally to the virus-treated SN for 2 weeks, before sacrifice and processing of brains for TH-immunohistochemistry. NGF treatment, in contrast to treatment with vehicle, significantly enhanced the number of dopaminergic neurons counted in the lesioned SN. These data suggest that ectopic TrkA can mediate the trophic actions of NGF and influence neuronal plasticity in vivo.</div></front></TEI><affiliations><list><country><li>Canada</li><li>France</li></country><region><li>Pays de la Loire</li></region><settlement><li>Nantes</li></settlement></list><tree><country name="France"><region name="Pays de la Loire"><name sortKey="Melchior, Benoit" sort="Melchior, Benoit" uniqKey="Melchior B" first="Benoit" last="Melchior">Benoit Melchior</name></region><name sortKey="Brachet, Philippe" sort="Brachet, Philippe" uniqKey="Brachet P" first="Philippe" last="Brachet">Philippe Brachet</name><name sortKey="Laplaud, David A" sort="Laplaud, David A" uniqKey="Laplaud D" first="David-A." last="Laplaud">David-A. Laplaud</name><name sortKey="Laplaud, David A" sort="Laplaud, David A" uniqKey="Laplaud D" first="David-A." last="Laplaud">David-A. Laplaud</name><name sortKey="Naveilhan, Philippe" sort="Naveilhan, Philippe" uniqKey="Naveilhan P" first="Philippe" last="Naveilhan">Philippe Naveilhan</name><name sortKey="Nerriere Daguin, Veronique" sort="Nerriere Daguin, Veronique" uniqKey="Nerriere Daguin V" first="Véronique" last="Nerriere-Daguin">Véronique Nerriere-Daguin</name><name sortKey="Neveu, Isabelle" sort="Neveu, Isabelle" uniqKey="Neveu I" first="Isabelle" last="Neveu">Isabelle Neveu</name><name sortKey="Remy, Severine" sort="Remy, Severine" uniqKey="Remy S" first="Séverine" last="Remy">Séverine Remy</name><name sortKey="Wiertlewski, Sandrine" sort="Wiertlewski, Sandrine" uniqKey="Wiertlewski S" first="Sandrine" last="Wiertlewski">Sandrine Wiertlewski</name></country><country name="Canada"><noRegion><name sortKey="Meakin, Susan O" sort="Meakin, Susan O" uniqKey="Meakin S" first="Susan O." last="Meakin">Susan O. Meakin</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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