Retroviral transfer of a human tyrosine hydroxylase cDNA in various cell lines: regulated release of dopamine in mouse anterior pituitary AtT-20 cells.
Identifieur interne : 001839 ( PubMed/Corpus ); précédent : 001838; suivant : 001840Retroviral transfer of a human tyrosine hydroxylase cDNA in various cell lines: regulated release of dopamine in mouse anterior pituitary AtT-20 cells.
Auteurs : P. Horellou ; B. Guibert ; V. Leviel ; J. MalletSource :
- Proceedings of the National Academy of Sciences of the United States of America [ 0027-8424 ] ; 1989.
English descriptors
- KwdEn :
- Animals, Aromatic-L-Amino-Acid Decarboxylases (metabolism), Catecholamines (biosynthesis), Catecholamines (secretion), Cell Line, Cells, Cultured, DNA (genetics), Dopamine (secretion), Genetic Vectors, Humans, Kinetics, Mice, Pituitary Neoplasms (secretion), Retroviridae (genetics), Transfection, Tyrosine 3-Monooxygenase (genetics), Tyrosine 3-Monooxygenase (metabolism).
- MESH :
- chemical , biosynthesis : Catecholamines.
- chemical , genetics : DNA, Tyrosine 3-Monooxygenase.
- chemical , metabolism : Aromatic-L-Amino-Acid Decarboxylases, Tyrosine 3-Monooxygenase.
- chemical , secretion : Catecholamines, Dopamine.
- genetics : Retroviridae.
- secretion : Pituitary Neoplasms.
- Animals, Cell Line, Cells, Cultured, Genetic Vectors, Humans, Kinetics, Mice, Transfection.
Abstract
Little is known about the molecular events mediating neurotransmitter release, a crucial step in synaptic transmission. In this paper, the biosynthesis and release of L-beta-3,4-dihydroxyphenylalanine (L-DOPA) and dopamine were analyzed in three heterologous cell lines after retroviral-mediated gene transfer of tyrosine hydroxylase (EC 1.14.16.2), the rate-limiting enzyme in catecholamine synthesis. A recombinant retrovirus encoding human tyrosine hydroxylase type I as well as neomycin-resistance gene was used to infect a fibroblast (NIH 3T3), a neuroblastoma (NS20 Y), and a neuroendocrine (AtT-20) cell line. After selection in the presence of neomycin and in tyrosine-free medium, high levels of exogenous tyrosine hydroxylase activity were detected in extracts of the three cell lines. High-performance liquid chromatography of cell extracts and culture supernatants confirmed that the three cell lines hydroxylated tyrosine to form L-DOPA and released this metabolite into the culture medium. Interestingly, the neuroendocrine cell line AtT-20 synthesized not only L-DOPA but also dopamine. Evoked secretion studies established that AtT-20 cells released the transmitter upon depolarization in a regulated, calcium-dependent way. We discuss the implication of this approach for the analyses of neurotransmitter release as well as in the context of degenerative disorders such as Parkinson disease.
PubMed: 2571152
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pubmed:2571152Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Retroviral transfer of a human tyrosine hydroxylase cDNA in various cell lines: regulated release of dopamine in mouse anterior pituitary AtT-20 cells.</title><author><name sortKey="Horellou, P" sort="Horellou, P" uniqKey="Horellou P" first="P" last="Horellou">P. Horellou</name><affiliation><nlm:affiliation>Laboratoire de Neurobiologie Cellulaire et Moléculaire, Centre National de la Recherche Scientifique, Gif-sur-Yvette, France.</nlm:affiliation></affiliation></author><author><name sortKey="Guibert, B" sort="Guibert, B" uniqKey="Guibert B" first="B" last="Guibert">B. Guibert</name></author><author><name sortKey="Leviel, V" sort="Leviel, V" uniqKey="Leviel V" first="V" last="Leviel">V. Leviel</name></author><author><name sortKey="Mallet, J" sort="Mallet, J" uniqKey="Mallet J" first="J" last="Mallet">J. Mallet</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="1989">1989</date><idno type="RBID">pubmed:2571152</idno><idno type="pmid">2571152</idno><idno type="wicri:Area/PubMed/Corpus">001839</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">001839</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Retroviral transfer of a human tyrosine hydroxylase cDNA in various cell lines: regulated release of dopamine in mouse anterior pituitary AtT-20 cells.</title><author><name sortKey="Horellou, P" sort="Horellou, P" uniqKey="Horellou P" first="P" last="Horellou">P. Horellou</name><affiliation><nlm:affiliation>Laboratoire de Neurobiologie Cellulaire et Moléculaire, Centre National de la Recherche Scientifique, Gif-sur-Yvette, France.</nlm:affiliation></affiliation></author><author><name sortKey="Guibert, B" sort="Guibert, B" uniqKey="Guibert B" first="B" last="Guibert">B. Guibert</name></author><author><name sortKey="Leviel, V" sort="Leviel, V" uniqKey="Leviel V" first="V" last="Leviel">V. Leviel</name></author><author><name sortKey="Mallet, J" sort="Mallet, J" uniqKey="Mallet J" first="J" last="Mallet">J. Mallet</name></author></analytic><series><title level="j">Proceedings of the National Academy of Sciences of the United States of America</title><idno type="ISSN">0027-8424</idno><imprint><date when="1989" type="published">1989</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Aromatic-L-Amino-Acid Decarboxylases (metabolism)</term><term>Catecholamines (biosynthesis)</term><term>Catecholamines (secretion)</term><term>Cell Line</term><term>Cells, Cultured</term><term>DNA (genetics)</term><term>Dopamine (secretion)</term><term>Genetic Vectors</term><term>Humans</term><term>Kinetics</term><term>Mice</term><term>Pituitary Neoplasms (secretion)</term><term>Retroviridae (genetics)</term><term>Transfection</term><term>Tyrosine 3-Monooxygenase (genetics)</term><term>Tyrosine 3-Monooxygenase (metabolism)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="biosynthesis" xml:lang="en"><term>Catecholamines</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>DNA</term><term>Tyrosine 3-Monooxygenase</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Aromatic-L-Amino-Acid Decarboxylases</term><term>Tyrosine 3-Monooxygenase</term></keywords><keywords scheme="MESH" type="chemical" qualifier="secretion" xml:lang="en"><term>Catecholamines</term><term>Dopamine</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Retroviridae</term></keywords><keywords scheme="MESH" qualifier="secretion" xml:lang="en"><term>Pituitary Neoplasms</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Cell Line</term><term>Cells, Cultured</term><term>Genetic Vectors</term><term>Humans</term><term>Kinetics</term><term>Mice</term><term>Transfection</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Little is known about the molecular events mediating neurotransmitter release, a crucial step in synaptic transmission. In this paper, the biosynthesis and release of L-beta-3,4-dihydroxyphenylalanine (L-DOPA) and dopamine were analyzed in three heterologous cell lines after retroviral-mediated gene transfer of tyrosine hydroxylase (EC 1.14.16.2), the rate-limiting enzyme in catecholamine synthesis. A recombinant retrovirus encoding human tyrosine hydroxylase type I as well as neomycin-resistance gene was used to infect a fibroblast (NIH 3T3), a neuroblastoma (NS20 Y), and a neuroendocrine (AtT-20) cell line. After selection in the presence of neomycin and in tyrosine-free medium, high levels of exogenous tyrosine hydroxylase activity were detected in extracts of the three cell lines. High-performance liquid chromatography of cell extracts and culture supernatants confirmed that the three cell lines hydroxylated tyrosine to form L-DOPA and released this metabolite into the culture medium. Interestingly, the neuroendocrine cell line AtT-20 synthesized not only L-DOPA but also dopamine. Evoked secretion studies established that AtT-20 cells released the transmitter upon depolarization in a regulated, calcium-dependent way. We discuss the implication of this approach for the analyses of neurotransmitter release as well as in the context of degenerative disorders such as Parkinson disease.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">2571152</PMID><DateCreated><Year>1989</Year><Month>10</Month><Day>24</Day></DateCreated><DateCompleted><Year>1989</Year><Month>10</Month><Day>24</Day></DateCompleted><DateRevised><Year>2013</Year><Month>11</Month><Day>21</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0027-8424</ISSN><JournalIssue CitedMedium="Print"><Volume>86</Volume><Issue>18</Issue><PubDate><Year>1989</Year><Month>Sep</Month></PubDate></JournalIssue><Title>Proceedings of the National Academy of Sciences of the United States of America</Title><ISOAbbreviation>Proc. Natl. Acad. Sci. U.S.A.</ISOAbbreviation></Journal><ArticleTitle>Retroviral transfer of a human tyrosine hydroxylase cDNA in various cell lines: regulated release of dopamine in mouse anterior pituitary AtT-20 cells.</ArticleTitle><Pagination><MedlinePgn>7233-7</MedlinePgn></Pagination><Abstract><AbstractText>Little is known about the molecular events mediating neurotransmitter release, a crucial step in synaptic transmission. In this paper, the biosynthesis and release of L-beta-3,4-dihydroxyphenylalanine (L-DOPA) and dopamine were analyzed in three heterologous cell lines after retroviral-mediated gene transfer of tyrosine hydroxylase (EC 1.14.16.2), the rate-limiting enzyme in catecholamine synthesis. A recombinant retrovirus encoding human tyrosine hydroxylase type I as well as neomycin-resistance gene was used to infect a fibroblast (NIH 3T3), a neuroblastoma (NS20 Y), and a neuroendocrine (AtT-20) cell line. After selection in the presence of neomycin and in tyrosine-free medium, high levels of exogenous tyrosine hydroxylase activity were detected in extracts of the three cell lines. High-performance liquid chromatography of cell extracts and culture supernatants confirmed that the three cell lines hydroxylated tyrosine to form L-DOPA and released this metabolite into the culture medium. Interestingly, the neuroendocrine cell line AtT-20 synthesized not only L-DOPA but also dopamine. Evoked secretion studies established that AtT-20 cells released the transmitter upon depolarization in a regulated, calcium-dependent way. We discuss the implication of this approach for the analyses of neurotransmitter release as well as in the context of degenerative disorders such as Parkinson disease.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Horellou</LastName><ForeName>P</ForeName><Initials>P</Initials><AffiliationInfo><Affiliation>Laboratoire de Neurobiologie Cellulaire et Moléculaire, Centre National de la Recherche Scientifique, Gif-sur-Yvette, France.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Guibert</LastName><ForeName>B</ForeName><Initials>B</Initials></Author><Author ValidYN="Y"><LastName>Leviel</LastName><ForeName>V</ForeName><Initials>V</Initials></Author><Author ValidYN="Y"><LastName>Mallet</LastName><ForeName>J</ForeName><Initials>J</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. 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Line</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D002478" MajorTopicYN="N">Cells, Cultured</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D004247" MajorTopicYN="N">DNA</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D004298" MajorTopicYN="N">Dopamine</DescriptorName><QualifierName UI="Q000557" MajorTopicYN="Y">secretion</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D005822" MajorTopicYN="N">Genetic Vectors</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D006801" MajorTopicYN="N">Humans</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D007700" MajorTopicYN="N">Kinetics</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D051379" MajorTopicYN="N">Mice</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D010911" MajorTopicYN="N">Pituitary Neoplasms</DescriptorName><QualifierName UI="Q000557" MajorTopicYN="N">secretion</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D012190" MajorTopicYN="N">Retroviridae</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D014162" MajorTopicYN="Y">Transfection</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D014446" MajorTopicYN="N">Tyrosine 3-Monooxygenase</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading></MeshHeadingList><OtherID Source="NLM">PMC298031</OtherID></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="pubmed"><Year>1989</Year><Month>9</Month><Day>1</Day></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>1989</Year><Month>9</Month><Day>1</Day><Hour>0</Hour><Minute>1</Minute></PubMedPubDate><PubMedPubDate 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