An Integrated Strategy for Analyzing the Unique Developmental Programs of Different Myoblast Subtypes 英文参考文献.docVIP
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An Integrated Strategy for Analyzing the Unique Developmental Programs of Different Myoblast Subtypes 英文参考文献
AnIntegratedStrategyforAnalyzing
theUniqueDevelopmentalPrograms
ofDifferentMyoblastSubtypes
Beatriz Estrada1,2[,Sung E.Choe1,3[,Stephen S.Gisselbrecht1,2,Sebastien Michaud1,2,¤a,Lakshmi Raj1,2¤b
Brian W. Busser1,2,Marc S.Halfon1¤c,George M.Church3,Alan M. Michelson1,2*
,
1DivisionofGenetics,DepartmentofMedicine,BrighamandWomen’sHospitalandHarvardMedicalSchool,Boston,Massachusetts,UnitedStatesofAmerica,2Howard
Hughes Medical Institute,Boston, Massachusetts, United States of America, 3 Department of Genetics, Harvard Medical School, Boston, Massachusetts, United States of
America
An important but largely unmet challenge in understanding the mechanisms that govern the formation of specific
organsistodecipherthecomplexanddynamicgeneticprogramsexhibitedbythediversityofcelltypeswithinthe
tissue of interest. Here, we use an integrated genetic, genomic, and computational strategy to comprehensively
determinethemolecularidentitiesofdistinctmyoblastsubpopulationswithintheDrosophilaembryonicmesodermat
thetimethatcellfatesareinitiallyspecified.Acompendiumofgeneexpressionprofileswasgeneratedforprimary
mesodermalcellspurifiedbyflowcytometryfromappropriatelystagedwild-typeembryosandfrom12genotypesin
which myogenesis was selectively and predictably perturbed. A statistical meta-analysis of these pooled datasets—
basedonexpectedtrendsingeneexpressionandontherelativecontributionofeachgenotypetothedetectionof
known muscle genes—provisionally assigned hundreds of differentially expressed genes to particular myoblast
subtypes.Wholeembryoinsituhybridizationswerethenusedtovalidatethemajorityofthesepredictions,thereby
enabling true-positive detection rates to be estimated for the microarray data. This combined analysis reveals that
myoblasts exhibit much greater gene expression heterogeneity and overall complexity than was previously
appreciated. Moreover, it implicates the involvement of large numbers of uncharacterized, differentially expressed
genes in myogenic specification and
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