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- W2775273957 abstract "Department of Anatomy and Cell BiologyUniversity of Kansas Medical Center3901 Rainbow BoulevardKansas City, Kansas 66160SummaryMicroRNAs (miRNAs) regulate gene expression bycontrollingtheturnover,translation,orbothofspecificmRNAs. In Drosophila, Dicer-1 (Dcr-1) is essential forgenerating mature miRNAs from their correspondingprecursors. Because miRNAs are known to modulatedevelopmental events, such as cell fate determinationand maintenance in many species, we investigatedwhether a lack of Dcr-1 would affect the maintenanceof stem cells (germline stem cells, GSCs; somaticstem cells, SSCs) in the Drosophila ovary by specifi-cally removing its function from the stem cells. Ourresults show that dcr-1 mutant GSCs cannot be main-tained and are lost rapidly from the niche without dis-cernable features of cell death, indicating that Dcr-1controlsGSCself-renewalbutnotsurvival.bagofmar-bles (bam), the gene that encodes an important differ-entiating factor in the Drosophila germline, however,is not upregulated in dcr-1 mutant GSCs, and itsremoval does not slow down dcr-1 mutant GSC loss,suggestingthatDcr-1controlsGSCself-renewalbyre-pressing a Bam-independent differentiation pathway.Furthermore, Dcr-1 is also essential for the mainte-nance of SSCs in the Drosophila ovary. Our data sug-gest that miRNAs produced by Dcr-1 are required formaintaining two types of stem cells in the Drosophilaovary.Results and DiscussionDuring miRNA biogenesis, Dcr-1 and its cofactor TRBP/Loquacious(Loqs)cleavethehairpinstructureofmiRNAprecursors into double-stranded RNAs (dsRNAs) [1–7],which are then further processed to generate maturemiRNAs.ThesemiRNAsplayimportantrolesindevelop-mental events, antiviral defense, and genome stability[8, 9]. For example, Dcr-1 null mice are embryonic lethalwith depletion of stem cells. It remains unclear whetherthe miRNA or small interference RNA (siRNA) pathwaycontrols the stem cell maintenance because the mouseDcr-1processesboth siRNAsandmiRNAs [10].The factthat two Dicers in Drosophila, Dcr-1 and Dcr-2, are re-quired for miRNA and siRNA pathways, respectively,makes Drosophila a unique system for dissectingfunctions of siRNAs and miRNAs in stem cells [2]. TheDrosophila ovaries contain three types of adult stemcells (germline stem cells [GSCs], escort stem cells[ESCs], and somatic stem cells [SSCs]), which producedifferentiated germ cells, escort cells, and somatic folli-cle cells, respectively, for life-long egg production [11–13] (Figures 1A and 1B). In the ovaries that are mutantfor a hypomorphic loqs mutation, GSCs are lost prema-turely, indicating that the miRNA pathway is required formaintaining GSCs [1, 3]. However, it remains unclearwhether Loqs functions inside the GSC to control itsmaintenance. Piwi, a known component in the miRNApathway, is required in the niche for controlling GSCself-renewal but is required intrinsically for GSC division[14–18]. One recent study further shows that Dcr-1 isrequired intrinsically to control GSC division only, butnot maintenance [19]. However, it remains uncertainwhether the miRNA pathway is required intrinsically forcontrollingthemaintenanceofanyofthestemcelltypesin the Drosophila ovary. Surprisingly, in this study, wedirectly show that Dcr-1 is required for the maintenanceof both GSCs and SSCs.GSCs in the Drosophila ovary can be identified easilybytheirphysicallocation(anteriorlyindirectcontactwithcapcells)andlocationofa sphericalorganelle knownasa spectrosome (anteriorly anchored close to cap cells)[12].ThesphericalspectrosomeinGSCsandtheirimme-diate daughters, known as cystoblasts, along with thebranch-shaped fusome in more mature cystocytes, canbe visualized by a molecular marker Hu-li tai-shao (Hts)[20]. To investigate whether dcr-1 is involved in GSCmaintenance in the Drosophila ovary, we used the FLP-mediated FRT recombination to generate marked dcr-1mutantGSCs.Theseareidentifiedasthemostanteriorlylocated LacZ-negative cells that also contain an anteri-orly localized spectrosome in the germaria (Figures1C–1K) according to our previously published proce-dures [21–23]. Along with the marked control wild-typeGSCs that were generated under identical conditions,these marked mutant dcr-1 GSCs were examined at dif-ferent time points (1 week, 2 weeks, and 3 weeks) afterclone induction (ACI) for determining the maintenanceratesofthesedcr-1mutantGSCsinacourseof3weeks.In the control, the percentage of the germaria with atleastone markedGSCdecreased from29.4%atthefirstweek ACI to 25.3% at the third week ACI (Table 1). Thisgradual loss reflects the normal GSC turnover, whichhas been reported independently in many publications[21, 22]. Most of the marked control GSCs detected atthe first week ACI still remained in the germarium at thesecond week (87.5%) and the third week (83.5%) ACI(Table 1; Figures 1C–1E). In contrast, for a strong dcr-1allele, dcr-1" @default.
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- W2775273957 date "2007-01-01" @default.
- W2775273957 modified "2023-09-23" @default.
- W2775273957 title "Report Dcr-1 Maintains Drosophila Ovarian Stem Cells" @default.
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