Abstract
ABSTRACTStem cells can self-renew and produce daughter cells destined for differentiation. The precise control of the balance between these two outcomes is essential to ensure tissue homeostasis and to prevent uncontrolled proliferation resulting in tumor formation. As self-renewal and differentiation are likely to be controlled by different gene expression programs, unraveling the underlying gene regulatory networks is crucial for understanding the molecular logic of this system. In this study, we have characterized by next generation RNA sequencing (RNA-seq) the transcriptome of germline stem cell (GSC)-like cells isolated from bag of marbles (bam) mutant Drosophila ovaries and compared it to the transcriptome of germ line cells isolated from wild-type ovaries. We have complemented this dataset by utilizing an RNA-immunoprecipitation strategy to identify transcripts bound to the master differentiation factor Bam. Protein complex enrichment analysis on these combined datasets allows us to delineate known and novel networks essential for GSC maintenance and differentiation. Further comparative transcriptomics illustrates similarities between GSCs and primordial germ cells and provides a molecular footprint of the stem cell state. Our study represents a useful resource for functional studies on stem cell maintenance and differentiation.
Highlights
Since being first isolated and in vitro propagated from mouse blastocysts in 1981, stem cells have piqued considerable scientific interest and captivated the society, albeit with a fair share of debate (Baylis and McLeod, 2007; Evans and Kaufman, 1981; McLaren, 2001; Watt and Driskell, 2010)
Mutation in bam causes a massive change in gene expression in the female germ line With only two or three germline stem cell (GSC) per ovariole, GSC gene expression profiling faces a challenge in that insufficient amounts of RNA might be available for sequencing
This problem can be attenuated by utilizing GSC-like cells which emerge as a consequence of bam mutation or Dpp overexpression (McKearin and Ohlstein, 1995; Xie and Spradling, 1998)
Summary
Since being first isolated and in vitro propagated from mouse blastocysts in 1981, stem cells have piqued considerable scientific interest and captivated the society, albeit with a fair share of debate (Baylis and McLeod, 2007; Evans and Kaufman, 1981; McLaren, 2001; Watt and Driskell, 2010). Received 5 August 2019; Accepted 16 October 2019 deterministically to renew themselves and produce progeny with restricted developmental potential (Morrison et al, 1997). Their hallmark self-renewal is essential for tissue maintenance in multicellular organisms and has for a long time held considerable promise for regenerative cell therapies (Singec et al, 2007). All this enthusiasm for stem cells has been propelled by advances in stem cell biology, which have been fueled and complemented by research on model organisms (Hunter, 2008). Following enclosure by somatic follicle cells, the cyst embarks on a maturation program, which culminates in the production of an egg ready for fertilization (Spradling et al, 2011)
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