Transcriptome analyses based on genetic screens for Pax3 myogenic targets in the mouse embryo
| Autor: | Ana Cumano, Margaret Buckingham, Frédéric Relaix, Jonathan D. Licht, Aimée Zuniga, Takahiko Sato, Mounia Lagha, Béatrice Regnault |
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| Přispěvatelé: | Génétique Moléculaire du Développement, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Génotypage des Eucaryotes (Plate-Forme), Institut Pasteur [Paris] (IP), Développement des Lymphocytes, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), DBM/Center for Biomedicine, Northwestern University [Evanston], Groupe Myologie, Institut de Myologie, Université Pierre et Marie Curie - Paris 6 (UPMC)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Association française contre les myopathies (AFM-Téléthon)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Association française contre les myopathies (AFM-Téléthon)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut de Génétique Moléculaire de Montpellier (IGMM), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris], Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Association française contre les myopathies (AFM-Téléthon)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Association française contre les myopathies (AFM-Téléthon)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM), BMC, Ed. |
| Jazyk: | angličtina |
| Rok vydání: | 2010 |
| Předmět: |
MESH: Signal Transduction
Transcription Genetic PAX3 Muscle Proteins MESH: Base Sequence Muscle Development Transcriptome Mice 0302 clinical medicine MESH: Gene Expression Regulation Developmental Paired Box Transcription Factors MESH: Animals Oligonucleotide Array Sequence Analysis Genetics 0303 health sciences MESH: Genetic Testing Forkhead Box Protein O1 Myogenesis Gene Expression Regulation Developmental PAX7 Transcription Factor Forkhead Transcription Factors musculoskeletal system MESH: Reproducibility of Results medicine.anatomical_structure MESH: Cell Survival embryonic structures [SDV.BBM.GTP] Life Sciences [q-bio]/Biochemistry Molecular Biology/Genomics [q-bio.GN] MESH: Muscle Development MYF5 Research Article Signal Transduction Biotechnology lcsh:QH426-470 Cell Survival lcsh:Biotechnology Mammalian/cytology/*metabolism Forkhead Transcription Factors/genetics/metabolism Gene Expression Profiling/*methods Gene Expression Regulation Biology Histone Deacetylases 03 medical and health sciences MESH: Muscle Proteins MESH: Gene Expression Profiling Genetic MESH: Forkhead Transcription Factors lcsh:TP248.13-248.65 [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry Molecular Biology/Genomics [q-bio.GN] medicine Animals Developmental *Genetic Testing Histone Deacetylases/genetics/metabolism Mice Muscle Development/*genetics Muscle Proteins/genetics/metabolism Oligonucleotide Array Sequence Analysis PAX7 Transcription Factor/genetics/metabolism Paired Box Transcription Factors/genetics/*metabolism RNA [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology Genetic Testing RNA Messenger MESH: Paired Box Transcription Factors Progenitor cell PAX3 Transcription Factor MESH: Mice 030304 developmental biology MESH: RNA Messenger Base Sequence Gene Expression Profiling Animals Base Sequence Cell Survival Embryo MESH: Transcription Genetic MESH: Embryo Mammalian Reproducibility of Results Messenger/genetics/metabolism Reproducibility of Results Signal Transduction/genetics Transcription Embryo Mammalian MESH: Histone Deacetylases lcsh:Genetics Somite MESH: PAX7 Transcription Factor MESH: Oligonucleotide Array Sequence Analysis PAX7 030217 neurology & neurosurgery Genetic screen |
| Zdroj: | BMC Genomics BMC Genomics, 2010, 11 (1), pp.696. ⟨10.1186/1471-2164-11-696⟩ BMC Genomics, BioMed Central, 2010, 11, pp.696. ⟨10.1186/1471-2164-11-696⟩ BMC Genomics, Vol 11, Iss 1, p 696 (2010) BMC Genomics; Vol 11 BMC Genomics, BioMed Central, 2010, 11 (1), pp.696. ⟨10.1186/1471-2164-11-696⟩ |
| ISSN: | 1471-2164 |
| DOI: | 10.1186/1471-2164-11-696⟩ |
| Popis: | Background Pax3 is a key upstream regulator of the onset of myogenesis, controlling progenitor cell survival and behaviour as well as entry into the myogenic programme. It functions in the dermomyotome of the somite from which skeletal muscle derives and in progenitor cell populations that migrate from the somite such as those of the limbs. Few Pax3 target genes have been identified. Identifying genes that lie genetically downstream of Pax3 is therefore an important endeavour in elucidating the myogenic gene regulatory network. Results We have undertaken a screen in the mouse embryo which employs a Pax3 GFP allele that permits isolation of Pax3 expressing cells by flow cytometry and a Pax3 PAX3-FKHR allele that encodes PAX3-FKHR in which the DNA binding domain of Pax3 is fused to the strong transcriptional activation domain of FKHR. This constitutes a gain of function allele that rescues the Pax3 mutant phenotype. Microarray comparisons were carried out between Pax3 GFP/+ and Pax3 GFP/PAX3-FKHR preparations from the hypaxial dermomyotome of somites at E9.5 and forelimb buds at E10.5. A further transcriptome comparison between Pax3-GFP positive and negative cells identified sequences specific to myogenic progenitors in the forelimb buds. Potential Pax3 targets, based on changes in transcript levels on the gain of function genetic background, were validated by analysis on loss or partial loss of function Pax3 mutant backgrounds. Sequences that are up- or down-regulated in the presence of PAX3-FKHR are classified as somite only, somite and limb or limb only. The latter should not contain sequences from Pax3 positive neural crest cells which do not invade the limbs. Verification by whole mount in situ hybridisation distinguishes myogenic markers. Presentation of potential Pax3 target genes focuses on signalling pathways and on transcriptional regulation. Conclusions Pax3 orchestrates many of the signalling pathways implicated in the activation or repression of myogenesis by regulating effectors and also, notably, inhibitors of these pathways. Important transcriptional regulators of myogenesis are candidate Pax3 targets. Myogenic determination genes, such as Myf5 are controlled positively, whereas the effect of Pax3 on genes encoding inhibitors of myogenesis provides a potential brake on differentiation. In the progenitor cell population, Pax7 and also Hdac5 which is a potential repressor of Foxc2, are subject to positive control by Pax3. |
| Databáze: | OpenAIRE |
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