A quantitative real-time RT-PCR assay for European eel tyrosine hydroxylase.
Dopamine (DA) plays a key inhibitory role in pubertal development of the European eel, but how DAergic neuronal activity is regulated is not known in this species. In order to investigate the regulation of DA inhibition at the molecular level, we developed a quantitative real-time RT-PCR (qrtRT-PCR)...
Published in: | General and Comparative Endocrinology |
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Online Access: | https://hal.science/hal-00018371 https://doi.org/10.1016/j.ygcen.2004.12.019 |
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ftsorbonneuniv:oai:HAL:hal-00018371v1 2024-09-15T18:05:14+00:00 A quantitative real-time RT-PCR assay for European eel tyrosine hydroxylase. Weltzien, Finn-Arne Pasqualini, Catherine Vernier, Philippe Dufour, Sylvie Biologie des organismes marins et écosystèmes (BOME) Muséum national d'Histoire naturelle (MNHN)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS) Développement, évolution et plasticité du système nerveux (DEPSN) Centre National de la Recherche Scientifique (CNRS) Institut de Neurobiologie Alfred Fessard (INAF) 2005 https://hal.science/hal-00018371 https://doi.org/10.1016/j.ygcen.2004.12.019 en eng HAL CCSD Elsevier info:eu-repo/semantics/altIdentifier/doi/10.1016/j.ygcen.2004.12.019 info:eu-repo/semantics/altIdentifier/pmid/15862557 hal-00018371 https://hal.science/hal-00018371 doi:10.1016/j.ygcen.2004.12.019 PUBMED: 15862557 ISSN: 0016-6480 EISSN: 1095-6840 General and Comparative Endocrinology https://hal.science/hal-00018371 General and Comparative Endocrinology, 2005, 142, pp.134. ⟨10.1016/j.ygcen.2004.12.019⟩ Phosphoproteins Ribosomal Proteins phosphoprotein P0 Cytochromes b Algorithms Amino Acid Sequence Anguilla metabolism Animals Brain anatomy & histology enzymology Cloning Molecular Cytochromes b genetics DNA Complementary biosynthesis genetics Female Molecular Sequence Data Phosphoproteins genetics metabolism RNA biosynthesis isolation & purification Reverse Transcriptase Polymerase Chain Reaction Ribosomal Proteins genetics metabolism Tyrosine 3-Monooxygenase [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC] info:eu-repo/semantics/article Journal articles 2005 ftsorbonneuniv https://doi.org/10.1016/j.ygcen.2004.12.019 2024-07-25T23:47:34Z Dopamine (DA) plays a key inhibitory role in pubertal development of the European eel, but how DAergic neuronal activity is regulated is not known in this species. In order to investigate the regulation of DA inhibition at the molecular level, we developed a quantitative real-time RT-PCR (qrtRT-PCR) assay, using the Light Cycler system, for the expression of eel tyrosine hydroxylase (TH), the rate-limiting enzyme in catecholamine biosynthesis. Two different reference genes were compared: the previously cloned eel cytochrome b, and eel acidic ribosomal phosphoprotein P0, the latter of which we cloned and partly sequenced. To further validate the assay, different methods of total RNA extraction were tested and compared. When applied to cDNA extracted from dissected brains of juvenile eels, the expression of TH was highest in the olfactory bulb, followed by the telencephalon including preoptic area, and the di-/mesencephalic areas excluding the optic lobes. TH expression in the optic lobes and in the medulla oblongata was low, whereas no expression could be detected in corpus cerebellum. This distribution pattern is in agreement with earlier studies on TH in the eel using immunohistochemistry, RT-PCR, and Northern blotting. The developed qrtRT-PCR assay provides a new tool for understanding the mechanisms regulating central DA inhibition of puberty in juvenile eels. Article in Journal/Newspaper European eel HAL Sorbonne Université General and Comparative Endocrinology 142 1-2 134 142 |
institution |
Open Polar |
collection |
HAL Sorbonne Université |
op_collection_id |
ftsorbonneuniv |
language |
English |
topic |
Phosphoproteins Ribosomal Proteins phosphoprotein P0 Cytochromes b Algorithms Amino Acid Sequence Anguilla metabolism Animals Brain anatomy & histology enzymology Cloning Molecular Cytochromes b genetics DNA Complementary biosynthesis genetics Female Molecular Sequence Data Phosphoproteins genetics metabolism RNA biosynthesis isolation & purification Reverse Transcriptase Polymerase Chain Reaction Ribosomal Proteins genetics metabolism Tyrosine 3-Monooxygenase [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC] |
spellingShingle |
Phosphoproteins Ribosomal Proteins phosphoprotein P0 Cytochromes b Algorithms Amino Acid Sequence Anguilla metabolism Animals Brain anatomy & histology enzymology Cloning Molecular Cytochromes b genetics DNA Complementary biosynthesis genetics Female Molecular Sequence Data Phosphoproteins genetics metabolism RNA biosynthesis isolation & purification Reverse Transcriptase Polymerase Chain Reaction Ribosomal Proteins genetics metabolism Tyrosine 3-Monooxygenase [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC] Weltzien, Finn-Arne Pasqualini, Catherine Vernier, Philippe Dufour, Sylvie A quantitative real-time RT-PCR assay for European eel tyrosine hydroxylase. |
topic_facet |
Phosphoproteins Ribosomal Proteins phosphoprotein P0 Cytochromes b Algorithms Amino Acid Sequence Anguilla metabolism Animals Brain anatomy & histology enzymology Cloning Molecular Cytochromes b genetics DNA Complementary biosynthesis genetics Female Molecular Sequence Data Phosphoproteins genetics metabolism RNA biosynthesis isolation & purification Reverse Transcriptase Polymerase Chain Reaction Ribosomal Proteins genetics metabolism Tyrosine 3-Monooxygenase [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC] |
description |
Dopamine (DA) plays a key inhibitory role in pubertal development of the European eel, but how DAergic neuronal activity is regulated is not known in this species. In order to investigate the regulation of DA inhibition at the molecular level, we developed a quantitative real-time RT-PCR (qrtRT-PCR) assay, using the Light Cycler system, for the expression of eel tyrosine hydroxylase (TH), the rate-limiting enzyme in catecholamine biosynthesis. Two different reference genes were compared: the previously cloned eel cytochrome b, and eel acidic ribosomal phosphoprotein P0, the latter of which we cloned and partly sequenced. To further validate the assay, different methods of total RNA extraction were tested and compared. When applied to cDNA extracted from dissected brains of juvenile eels, the expression of TH was highest in the olfactory bulb, followed by the telencephalon including preoptic area, and the di-/mesencephalic areas excluding the optic lobes. TH expression in the optic lobes and in the medulla oblongata was low, whereas no expression could be detected in corpus cerebellum. This distribution pattern is in agreement with earlier studies on TH in the eel using immunohistochemistry, RT-PCR, and Northern blotting. The developed qrtRT-PCR assay provides a new tool for understanding the mechanisms regulating central DA inhibition of puberty in juvenile eels. |
author2 |
Biologie des organismes marins et écosystèmes (BOME) Muséum national d'Histoire naturelle (MNHN)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS) Développement, évolution et plasticité du système nerveux (DEPSN) Centre National de la Recherche Scientifique (CNRS) Institut de Neurobiologie Alfred Fessard (INAF) |
format |
Article in Journal/Newspaper |
author |
Weltzien, Finn-Arne Pasqualini, Catherine Vernier, Philippe Dufour, Sylvie |
author_facet |
Weltzien, Finn-Arne Pasqualini, Catherine Vernier, Philippe Dufour, Sylvie |
author_sort |
Weltzien, Finn-Arne |
title |
A quantitative real-time RT-PCR assay for European eel tyrosine hydroxylase. |
title_short |
A quantitative real-time RT-PCR assay for European eel tyrosine hydroxylase. |
title_full |
A quantitative real-time RT-PCR assay for European eel tyrosine hydroxylase. |
title_fullStr |
A quantitative real-time RT-PCR assay for European eel tyrosine hydroxylase. |
title_full_unstemmed |
A quantitative real-time RT-PCR assay for European eel tyrosine hydroxylase. |
title_sort |
quantitative real-time rt-pcr assay for european eel tyrosine hydroxylase. |
publisher |
HAL CCSD |
publishDate |
2005 |
url |
https://hal.science/hal-00018371 https://doi.org/10.1016/j.ygcen.2004.12.019 |
genre |
European eel |
genre_facet |
European eel |
op_source |
ISSN: 0016-6480 EISSN: 1095-6840 General and Comparative Endocrinology https://hal.science/hal-00018371 General and Comparative Endocrinology, 2005, 142, pp.134. ⟨10.1016/j.ygcen.2004.12.019⟩ |
op_relation |
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.ygcen.2004.12.019 info:eu-repo/semantics/altIdentifier/pmid/15862557 hal-00018371 https://hal.science/hal-00018371 doi:10.1016/j.ygcen.2004.12.019 PUBMED: 15862557 |
op_doi |
https://doi.org/10.1016/j.ygcen.2004.12.019 |
container_title |
General and Comparative Endocrinology |
container_volume |
142 |
container_issue |
1-2 |
container_start_page |
134 |
op_container_end_page |
142 |
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1810442823871234048 |