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ES-cells Carrying Two Inactivated myf-5 Alleles Form Skeletal Muscle Cells: Activation of an Alternative myf-5-Independent Differentiation Pathway

机译:携带两个灭活的myf-5等位基因的ES细胞形成骨骼肌细胞:替代myf-5独立分化途径的激活。

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Bothallelesofthemyogenicregulatorygenemyf-5havebeeninactivatedinmouseembryonicstemcellsbydifferentstrategiesinvolvingeitherconsecutivegenetargetingwithneomycinandhygromycinreplacementvectorsorspontaneouslossofheterozygosityincellstargetedwiththeneomycinreplacementvectoralone.Bothselectionschemesprovidedhomozygousmyf-5mutantES-cellswithnormaldevelopmentalpotentialeminvitro/em.Embryoidbodiesdifferentiatedintoskeletalmusclecellsasassessedbytheirtypicalmorphologyandskeletalmusclemarkers.Theextentofdifferentiationinhomozygousmutantmyf-5embryonicstemcellswasvirtuallyindistinguishablefromcontrolcultures,suggestingthatmyf-5isnotrequiredfortheearlystepsofmyogenicdevelopmenteminvitro/em.Whilemyocytepopulationsderivedfromwild-typeandheterozygousmyf-5mutantES-cellscontainedmyoD-positiveandmyoD-negativecells,nomyoD-negativemusclecellswerefoundamongmyf-5homozygousmutants.Differentiatedmyf-5double-knockoutcellsalsoshowedaprematureexpressionofmyoD.TheseresultsindicateacompensatoryroleofmyoDandmyf-5duringearlymyocytedevelopmentandsuggestapossibledown-regulationofmyoDbymyf-5duringearlymyogenesis./p/div
机译:Bothallelesofthemyogenicregulatorygenemyf-5havebeeninactivatedinmouseembryonicstemcellsbydifferentstrategiesinvolvingeitherconsecutivegenetargetingwithneomycinandhygromycinreplacementvectorsorspontaneouslossofheterozygosityincellstargetedwiththeneomycinreplacementvectoralone.Bothselectionschemesprovidedhomozygousmyf-5mutantES-cellswithnormaldevelopmentalpotential 体外 .Embryoidbodiesdifferentiatedintoskeletalmusclecellsasassessedbytheirtypicalmorphologyandskeletalmusclemarkers.Theextentofdifferentiationinhomozygousmutantmyf-5embryonicstemcellswasvirtuallyindistinguishablefromcontrolcultures,suggestingthatmyf-5isnotrequiredfortheearlystepsofmyogenicdevelopment 体外 .Whilemyocytepopulationsderivedfromwild-typeandheterozygousmyf-5mutantES-cellscontainedmyoD-positiveandmyoD-negativecells,nomyoD-在myf-5纯合突变体中发现了阴性肌肉细胞。分化的myf-5双敲除细胞还显示了myoD的过早表达。早期心肌细胞发育过程中的myf-5和早期成肌过程中myf-5对myoD的可能下调。

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