Uptake of antisense oligonucleotides and functional block of acetylcholine receptor subunit gene expression in primary embryonic neurons.
Identifieur interne : 002917 ( Ncbi/Curation ); précédent : 002916; suivant : 002918Uptake of antisense oligonucleotides and functional block of acetylcholine receptor subunit gene expression in primary embryonic neurons.
Auteurs : C. Yu [États-Unis] ; A B Brussaard ; X. Yang ; M. Listerud ; L W RoleSource :
- Developmental genetics [ 0192-253X ] ; 1993.
Descripteurs français
- KwdFr :
- Animaux, Canaux ioniques (génétique), Canaux ioniques (métabolisme), Cellules cultivées, Données de séquences moléculaires, Embryon de poulet, Endocytose, Ganglions sympathiques (cytologie), Ganglions sympathiques (embryologie), Neurones (), Neurones (métabolisme), Oligonucléotides antisens (génétique), Oligonucléotides antisens (métabolisme), Oligonucléotides antisens (pharmacologie), Récepteurs nicotiniques (biosynthèse), Récepteurs nicotiniques (génétique), Régulation négative, Séquence nucléotidique, Température, Transport biologique.
- MESH :
- biosynthèse : Récepteurs nicotiniques.
- cytologie : Ganglions sympathiques.
- embryologie : Ganglions sympathiques.
- génétique : Canaux ioniques, Oligonucléotides antisens, Récepteurs nicotiniques.
- métabolisme : Canaux ioniques, Neurones, Oligonucléotides antisens.
- pharmacologie : Oligonucléotides antisens.
- Animaux, Cellules cultivées, Données de séquences moléculaires, Embryon de poulet, Endocytose, Neurones, Régulation négative, Séquence nucléotidique, Température, Transport biologique.
English descriptors
- KwdEn :
- Animals, Base Sequence, Biological Transport, Cells, Cultured, Chick Embryo, Down-Regulation, Endocytosis, Ganglia, Sympathetic (cytology), Ganglia, Sympathetic (embryology), Ion Channels (genetics), Ion Channels (metabolism), Molecular Sequence Data, Neurons (drug effects), Neurons (metabolism), Oligonucleotides, Antisense (genetics), Oligonucleotides, Antisense (metabolism), Oligonucleotides, Antisense (pharmacology), Receptors, Nicotinic (biosynthesis), Receptors, Nicotinic (genetics), Temperature.
- MESH :
- chemical , biosynthesis : Receptors, Nicotinic.
- chemical , genetics : Ion Channels, Oligonucleotides, Antisense, Receptors, Nicotinic.
- cytology : Ganglia, Sympathetic.
- drug effects : Neurons.
- embryology : Ganglia, Sympathetic.
- chemical , metabolism : Ion Channels, Neurons, Oligonucleotides, Antisense.
- chemical , pharmacology : Oligonucleotides, Antisense.
- Animals, Base Sequence, Biological Transport, Cells, Cultured, Chick Embryo, Down-Regulation, Endocytosis, Molecular Sequence Data, Temperature.
Abstract
Several recent studies have used antisense oligonucleotides in the nervous system to probe the functional role of particular gene products. Since antisense oligonucleotide-mediated block of gene expression typically involves uptake of the oligonucleotides, we have characterized the mechanism of this uptake into developing neurons from embryonic chickens. Antisense oligonucleotides (15 mers) added to the bathing media are taken up into the embryonic chicken sympathetic neurons maintained in vitro. A portion of the oligonucleotide uptake is temperature dependent and saturates at extracellular oligonucleotide concentrations > or = 20 microM. This temperature sensitive, saturable component is effectively completed by single nucleotides of ATP and AMP and is reminiscent of receptor-mediated endocytosis of oligonucleotides described in non-neuronal cells. The efficiency of the oligonucleotide uptake system is dependent on the developmental stage of the animal but independent of the number of days that the neurons are maintained in vitro. Following the uptake of antisense oligonucleotides directed against ion channel subunit genes expressed by these neurons (nicotinic acetylcholine receptor subunit alpha 3; nAChR alpha 3), biophysical assays reveal that the functional expression of the target gene is largely blocked. Thus the number of wild type nAChR channels expressed is decreased by approximately 80%-90%. Furthermore, following antisense deletion of alpha 3, "mutant" nAChRs with distinct functional characteristics are expressed. In sum, these studies characterize the uptake of antisense oligonucleotide and demonstrate the functional block of specific gene expression in primary developing neurons.(ABSTRACT TRUNCATED AT 250 WORDS)
DOI: 10.1002/dvg.1020140407
PubMed: 7693387
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Yu</name><affiliation wicri:level="2"><nlm:affiliation>Department of Anatomy and Cell Biology, Center for Neurobiology and Behavior, College of Physicians and Surgeons, Columbia University, New York, New York 10032.</nlm:affiliation><country xml:lang="fr">États-Unis</country><placeName><region type="state">État de New York</region></placeName><wicri:cityArea>Department of Anatomy and Cell Biology, Center for Neurobiology and Behavior, College of Physicians and Surgeons, Columbia University, New York</wicri:cityArea></affiliation></author><author><name sortKey="Brussaard, A B" sort="Brussaard, A B" uniqKey="Brussaard A" first="A B" last="Brussaard">A B Brussaard</name></author><author><name sortKey="Yang, X" sort="Yang, X" uniqKey="Yang X" first="X" last="Yang">X. Yang</name></author><author><name sortKey="Listerud, M" sort="Listerud, M" uniqKey="Listerud M" first="M" last="Listerud">M. 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Yu</name><affiliation wicri:level="2"><nlm:affiliation>Department of Anatomy and Cell Biology, Center for Neurobiology and Behavior, College of Physicians and Surgeons, Columbia University, New York, New York 10032.</nlm:affiliation><country xml:lang="fr">États-Unis</country><placeName><region type="state">État de New York</region></placeName><wicri:cityArea>Department of Anatomy and Cell Biology, Center for Neurobiology and Behavior, College of Physicians and Surgeons, Columbia University, New York</wicri:cityArea></affiliation></author><author><name sortKey="Brussaard, A B" sort="Brussaard, A B" uniqKey="Brussaard A" first="A B" last="Brussaard">A B Brussaard</name></author><author><name sortKey="Yang, X" sort="Yang, X" uniqKey="Yang X" first="X" last="Yang">X. Yang</name></author><author><name sortKey="Listerud, M" sort="Listerud, M" uniqKey="Listerud M" first="M" last="Listerud">M. Listerud</name></author><author><name sortKey="Role, L W" sort="Role, L W" uniqKey="Role L" first="L W" last="Role">L W Role</name></author></analytic><series><title level="j">Developmental genetics</title><idno type="ISSN">0192-253X</idno><imprint><date when="1993" type="published">1993</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Base Sequence</term><term>Biological Transport</term><term>Cells, Cultured</term><term>Chick Embryo</term><term>Down-Regulation</term><term>Endocytosis</term><term>Ganglia, Sympathetic (cytology)</term><term>Ganglia, Sympathetic (embryology)</term><term>Ion Channels (genetics)</term><term>Ion Channels (metabolism)</term><term>Molecular Sequence Data</term><term>Neurons (drug effects)</term><term>Neurons (metabolism)</term><term>Oligonucleotides, Antisense (genetics)</term><term>Oligonucleotides, Antisense (metabolism)</term><term>Oligonucleotides, Antisense (pharmacology)</term><term>Receptors, Nicotinic (biosynthesis)</term><term>Receptors, Nicotinic (genetics)</term><term>Temperature</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Animaux</term><term>Canaux ioniques (génétique)</term><term>Canaux ioniques (métabolisme)</term><term>Cellules cultivées</term><term>Données de séquences moléculaires</term><term>Embryon de poulet</term><term>Endocytose</term><term>Ganglions sympathiques (cytologie)</term><term>Ganglions sympathiques (embryologie)</term><term>Neurones ()</term><term>Neurones (métabolisme)</term><term>Oligonucléotides antisens (génétique)</term><term>Oligonucléotides antisens (métabolisme)</term><term>Oligonucléotides antisens (pharmacologie)</term><term>Récepteurs nicotiniques (biosynthèse)</term><term>Récepteurs nicotiniques (génétique)</term><term>Régulation négative</term><term>Séquence nucléotidique</term><term>Température</term><term>Transport biologique</term></keywords><keywords scheme="MESH" type="chemical" qualifier="biosynthesis" xml:lang="en"><term>Receptors, Nicotinic</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Ion Channels</term><term>Oligonucleotides, Antisense</term><term>Receptors, Nicotinic</term></keywords><keywords scheme="MESH" qualifier="biosynthèse" xml:lang="fr"><term>Récepteurs nicotiniques</term></keywords><keywords scheme="MESH" qualifier="cytologie" xml:lang="fr"><term>Ganglions sympathiques</term></keywords><keywords scheme="MESH" qualifier="cytology" xml:lang="en"><term>Ganglia, Sympathetic</term></keywords><keywords scheme="MESH" qualifier="drug effects" xml:lang="en"><term>Neurons</term></keywords><keywords scheme="MESH" qualifier="embryologie" xml:lang="fr"><term>Ganglions sympathiques</term></keywords><keywords scheme="MESH" qualifier="embryology" xml:lang="en"><term>Ganglia, Sympathetic</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Canaux ioniques</term><term>Oligonucléotides antisens</term><term>Récepteurs nicotiniques</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Ion Channels</term><term>Neurons</term><term>Oligonucleotides, Antisense</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Canaux ioniques</term><term>Neurones</term><term>Oligonucléotides antisens</term></keywords><keywords scheme="MESH" qualifier="pharmacologie" xml:lang="fr"><term>Oligonucléotides antisens</term></keywords><keywords scheme="MESH" type="chemical" qualifier="pharmacology" xml:lang="en"><term>Oligonucleotides, Antisense</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Base Sequence</term><term>Biological Transport</term><term>Cells, Cultured</term><term>Chick Embryo</term><term>Down-Regulation</term><term>Endocytosis</term><term>Molecular Sequence Data</term><term>Temperature</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><term>Cellules cultivées</term><term>Données de séquences moléculaires</term><term>Embryon de poulet</term><term>Endocytose</term><term>Neurones</term><term>Régulation négative</term><term>Séquence nucléotidique</term><term>Température</term><term>Transport biologique</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Several recent studies have used antisense oligonucleotides in the nervous system to probe the functional role of particular gene products. Since antisense oligonucleotide-mediated block of gene expression typically involves uptake of the oligonucleotides, we have characterized the mechanism of this uptake into developing neurons from embryonic chickens. Antisense oligonucleotides (15 mers) added to the bathing media are taken up into the embryonic chicken sympathetic neurons maintained in vitro. A portion of the oligonucleotide uptake is temperature dependent and saturates at extracellular oligonucleotide concentrations > or = 20 microM. This temperature sensitive, saturable component is effectively completed by single nucleotides of ATP and AMP and is reminiscent of receptor-mediated endocytosis of oligonucleotides described in non-neuronal cells. The efficiency of the oligonucleotide uptake system is dependent on the developmental stage of the animal but independent of the number of days that the neurons are maintained in vitro. Following the uptake of antisense oligonucleotides directed against ion channel subunit genes expressed by these neurons (nicotinic acetylcholine receptor subunit alpha 3; nAChR alpha 3), biophysical assays reveal that the functional expression of the target gene is largely blocked. Thus the number of wild type nAChR channels expressed is decreased by approximately 80%-90%. Furthermore, following antisense deletion of alpha 3, "mutant" nAChRs with distinct functional characteristics are expressed. In sum, these studies characterize the uptake of antisense oligonucleotide and demonstrate the functional block of specific gene expression in primary developing neurons.(ABSTRACT TRUNCATED AT 250 WORDS)</div></front></TEI></record>Pour manipuler ce document sous Unix (Dilib)
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