title: FACS purification and transcriptome analysis of drosophila neural stem cells reveals a role for Klumpfuss in self-renewal.
abstract: Drosophila neuroblasts (NBs) have emerged as a model for stem cell biology that is ideal for genetic analysis but is limited by the lack of cell-type-specific gene expression data. Here, we describe a method for isolating large numbers of pure NBs and differentiating neurons that retain both cell-cycle and lineage characteristics. We determine transcriptional profiles by mRNA sequencing and identify 28 predicted NB-specific transcription factors that can be arranged in a network containing hubs for Notch signaling, growth control, and chromatin regulation. Overexpression and RNA interference for these factors identify Klumpfuss as a regulator of self-renewal. We show that loss of Klumpfuss function causes premature differentiation and that overexpression results in the formation of transplantable brain tumors. Our data represent a valuable resource for investigating Drosophila developmental neurobiology, and the described method can be applied to other invertebrate stem cell lineages as well.
authors.affiliation: Institute of Molecular Biotechnology of the Austrian Academy of Science, Vienna, Austria.
authors.forename: Christian
authors.initials: C
authors.lastname: Berger
authors.forename: Heike
authors.initials: H
authors.lastname: Harzer
authors.forename: Thomas R
authors.initials: TR
authors.lastname: Burkard
authors.forename: Jonas
authors.initials: J
authors.lastname: Steinmann
authors.forename: Suzanne
authors.initials: S
authors.lastname: van der Horst
authors.forename: Anne-Sophie
authors.initials: AS
authors.lastname: Laurenson
authors.forename: Maria
authors.initials: M
authors.lastname: Novatchkova
authors.forename: Heinrich
authors.initials: H
authors.lastname: Reichert
authors.forename: Juergen A
authors.initials: JA
authors.lastname: Knoblich
mesh_terms: D000818:Animals; D002453:Cell Cycle / Q000502:physiology*; D002454:Cell Differentiation / Q000502:physiology*; D019070:Cell Lineage / Q000502:physiology; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D004331:Drosophila melanogaster; D005434:Flow Cytometry / Q000379:methods; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000502:physiology*; D058953:Neural Stem Cells / Q000166:cytology / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D059467:Transcriptome / Q000502:physiology*
chemical_list: D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; C107732:Klu protein, Drosophila; D014157:Transcription Factors