Identification of endogenous changes in the transcriptome of young versus old Drosophila intestinal stem cells using Next Generation RNA Sequencing

The Drosophila midgut is an ideal model system to study molecular mechanisms that interfere with the intestinal stem cells’ (ISCs) ability to function in tissue homeostasis. Due to the lack of a combination of molecular markers suitable to isolate ISCs from aged intestines, it has been a major challenge to study endogenous molecular changes of ISCs during aging. Our FACS-based approach using the esg-GAL4, UAS-GFP fly line allowed the isolation of a cell population enriched for ISCs from young and old midguts by their small size, little granularity and low GFP intensity. The isolated ISCs were subsequently used for RNA sequencing to identify endogenous changes in the transcriptome of young versus old ISCs. Overall design: Cell populations enriched for ISCs isolated from young (6-8 days old) and old (59-65 days old) midguts were sorted. Cells from three different batches of young and old midguts were subjected to Next Generation Sequencing using Illumina Genome Analyzer IIx.

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Tauc HM, Tasdogan A, Meyer P, Pandur P