MyoD Cannot Compensate for the Absence of Myogenin during Skeletal Muscle Differentiation in Murine Embryonic Stem Cells

myogenin (−/−) mice display severe skeletal muscle defects despite expressing normal levels of MyoD. The failure of MyoD to compensate for myogenin could be explained by distinctions in protein function or by differences in patterns of gene expression. To distinguish between these two possibilities,...

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Veröffentlicht in:Developmental biology 2001-01, Vol.229 (2), p.340-350
Hauptverfasser: Myer, Anita, Olson, Eric N., Klein, William H.
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description myogenin (−/−) mice display severe skeletal muscle defects despite expressing normal levels of MyoD. The failure of MyoD to compensate for myogenin could be explained by distinctions in protein function or by differences in patterns of gene expression. To distinguish between these two possibilities, we compared the abilities of constitutively expressed myogenin and MyoD to support muscle differentiation in embryoid bodies made from myogenin (−/−) ES cells. Differentiated embryoid bodies from wild-type embryonic stem (ES) cells made extensive skeletal muscle, but embryoid bodies from myogenin (−/−) ES cells had greatly attenuated muscle-forming capacity. The inability of myogenin (−/−) ES cells to generate muscle was independent of endogenous MyoD expression. Skeletal muscle was restored in myogenin (−/−) ES cells by constitutive expression of myogenin. In contrast, constitutive expression of MyoD resulted in only marginal enhancement of skeletal muscle, although myocyte numbers greatly increased. The results indicated that constitutive expression of MyoD led to enhanced myogenic commitment of myogenin (−/−) cells but also indicated that committed cells were impaired in their ability to form muscle sheets without myogenin. Thus, despite their relatedness, myogenin's role in muscle formation is distinct from that of MyoD, and the distinction cannot be explained merely by differences in their expression properties.
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The failure of MyoD to compensate for myogenin could be explained by distinctions in protein function or by differences in patterns of gene expression. To distinguish between these two possibilities, we compared the abilities of constitutively expressed myogenin and MyoD to support muscle differentiation in embryoid bodies made from myogenin (−/−) ES cells. Differentiated embryoid bodies from wild-type embryonic stem (ES) cells made extensive skeletal muscle, but embryoid bodies from myogenin (−/−) ES cells had greatly attenuated muscle-forming capacity. The inability of myogenin (−/−) ES cells to generate muscle was independent of endogenous MyoD expression. Skeletal muscle was restored in myogenin (−/−) ES cells by constitutive expression of myogenin. In contrast, constitutive expression of MyoD resulted in only marginal enhancement of skeletal muscle, although myocyte numbers greatly increased. The results indicated that constitutive expression of MyoD led to enhanced myogenic commitment of myogenin (−/−) cells but also indicated that committed cells were impaired in their ability to form muscle sheets without myogenin. 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The failure of MyoD to compensate for myogenin could be explained by distinctions in protein function or by differences in patterns of gene expression. To distinguish between these two possibilities, we compared the abilities of constitutively expressed myogenin and MyoD to support muscle differentiation in embryoid bodies made from myogenin (−/−) ES cells. Differentiated embryoid bodies from wild-type embryonic stem (ES) cells made extensive skeletal muscle, but embryoid bodies from myogenin (−/−) ES cells had greatly attenuated muscle-forming capacity. The inability of myogenin (−/−) ES cells to generate muscle was independent of endogenous MyoD expression. Skeletal muscle was restored in myogenin (−/−) ES cells by constitutive expression of myogenin. In contrast, constitutive expression of MyoD resulted in only marginal enhancement of skeletal muscle, although myocyte numbers greatly increased. The results indicated that constitutive expression of MyoD led to enhanced myogenic commitment of myogenin (−/−) cells but also indicated that committed cells were impaired in their ability to form muscle sheets without myogenin. 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Olson, Eric N. ; Klein, William H.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c380t-d648a024831e0d3dca3b0ef58ea4094c185c348b6cba2efed8b92f8636c539cd3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2001</creationdate><topic>Animals</topic><topic>bHLH transcription factors</topic><topic>Cell Differentiation</topic><topic>Cell Division</topic><topic>Electroporation</topic><topic>ES cell differentiation</topic><topic>Gene Expression Regulation, Developmental</topic><topic>Helix-Loop-Helix Motifs</topic><topic>Mice</topic><topic>Mice, Knockout</topic><topic>Muscle, Skeletal - cytology</topic><topic>Muscle, Skeletal - embryology</topic><topic>MyoD</topic><topic>MyoD Protein - genetics</topic><topic>MyoD Protein - physiology</topic><topic>myogenin</topic><topic>Myogenin - genetics</topic><topic>Myogenin - physiology</topic><topic>Myosin Heavy Chains - analysis</topic><topic>Myosin Heavy Chains - genetics</topic><topic>Reverse Transcriptase Polymerase Chain Reaction</topic><topic>skeletal muscle formation</topic><topic>Stem Cells - cytology</topic><topic>Stem Cells - physiology</topic><topic>Transcription, Genetic</topic><topic>Transfection</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Myer, Anita</creatorcontrib><creatorcontrib>Olson, Eric N.</creatorcontrib><creatorcontrib>Klein, William H.</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Developmental biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Myer, Anita</au><au>Olson, Eric N.</au><au>Klein, William H.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>MyoD Cannot Compensate for the Absence of Myogenin during Skeletal Muscle Differentiation in Murine Embryonic Stem Cells</atitle><jtitle>Developmental biology</jtitle><addtitle>Dev Biol</addtitle><date>2001-01-15</date><risdate>2001</risdate><volume>229</volume><issue>2</issue><spage>340</spage><epage>350</epage><pages>340-350</pages><issn>0012-1606</issn><eissn>1095-564X</eissn><abstract>myogenin (−/−) mice display severe skeletal muscle defects despite expressing normal levels of MyoD. 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The results indicated that constitutive expression of MyoD led to enhanced myogenic commitment of myogenin (−/−) cells but also indicated that committed cells were impaired in their ability to form muscle sheets without myogenin. Thus, despite their relatedness, myogenin's role in muscle formation is distinct from that of MyoD, and the distinction cannot be explained merely by differences in their expression properties.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>11203698</pmid><doi>10.1006/dbio.2000.9985</doi><tpages>11</tpages><oa>free_for_read</oa></addata></record>
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subjects Animals
bHLH transcription factors
Cell Differentiation
Cell Division
Electroporation
ES cell differentiation
Gene Expression Regulation, Developmental
Helix-Loop-Helix Motifs
Mice
Mice, Knockout
Muscle, Skeletal - cytology
Muscle, Skeletal - embryology
MyoD
MyoD Protein - genetics
MyoD Protein - physiology
myogenin
Myogenin - genetics
Myogenin - physiology
Myosin Heavy Chains - analysis
Myosin Heavy Chains - genetics
Reverse Transcriptase Polymerase Chain Reaction
skeletal muscle formation
Stem Cells - cytology
Stem Cells - physiology
Transcription, Genetic
Transfection
title MyoD Cannot Compensate for the Absence of Myogenin during Skeletal Muscle Differentiation in Murine Embryonic Stem Cells
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