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The bantam microRNA is associated with drosophila fragile X mental retardation protein and regulates the fate of germline stem cells.

Yang Y, Xu S, Xia L, Wang J, Wen S, Jin P, Chen D - PLoS Genet. (2009)

Bottom Line: We show that, like dFmr1, bantam is not only required for repressing primordial germ cell differentiation, it also functions as an extrinsic factor for germline stem cell maintenance.Furthermore, we find that bantam genetically interacts with dFmr1 to regulate the fate of germline stem cells.Collectively, our results support the notion that the FMRP-mediated translation pathway functions through specific miRNAs to control stem cell regulation.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, People's Republic of China.

ABSTRACT
Fragile X syndrome, a common form of inherited mental retardation, is caused by the loss of fragile X mental retardation protein (FMRP). We have previously demonstrated that dFmr1, the Drosophila ortholog of the fragile X mental retardation 1 gene, plays a role in the proper maintenance of germline stem cells in Drosophila ovary; however, the molecular mechanism behind this remains elusive. In this study, we used an immunoprecipitation assay to reveal that specific microRNAs (miRNAs), particularly the bantam miRNA (bantam), are physically associated with dFmrp in ovary. We show that, like dFmr1, bantam is not only required for repressing primordial germ cell differentiation, it also functions as an extrinsic factor for germline stem cell maintenance. Furthermore, we find that bantam genetically interacts with dFmr1 to regulate the fate of germline stem cells. Collectively, our results support the notion that the FMRP-mediated translation pathway functions through specific miRNAs to control stem cell regulation.

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The bantam miRNA plays a non-autonomous role in GSC maintenance.GSC clones were induced by heat-shock treatment in adult female flies. Ovaries from FRT control flies (A and B) and FRT, ban flies (C and D) were dissected at day 2 and day 12 following heat-shock treatment; GSC clones were identified by the lack of GFP expression. Scale bar represents 10 µm. (E) Relative percentages of negatively GFP-marked GSC clones in FRT control and two FRT; ban  alleles at days 2, 7, 12, and 15 AHST are shown. P>0.5.
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pgen-1000444-g005: The bantam miRNA plays a non-autonomous role in GSC maintenance.GSC clones were induced by heat-shock treatment in adult female flies. Ovaries from FRT control flies (A and B) and FRT, ban flies (C and D) were dissected at day 2 and day 12 following heat-shock treatment; GSC clones were identified by the lack of GFP expression. Scale bar represents 10 µm. (E) Relative percentages of negatively GFP-marked GSC clones in FRT control and two FRT; ban alleles at days 2, 7, 12, and 15 AHST are shown. P>0.5.

Mentions: The loss of germline stem cells in ban mutant ovaries indicates that ban is required by either GSCs or somatic cells. To analyze whether ban functions as a cell-autonomous factor in maintaining GSC fate, we used a FLP-FRT–mediated mitotic recombination technique to generate marked mutant GSCs [17],[22], then calculated the life span of the marked mutant GSCs by quantifying their loss rate. The marked mutant GSCs were identified by a lack of GFP fluorescence in the nuclei and by their positions directly attaching to the base cells of the terminal filament or cap cells. The ban loss-of-function alleles (ban20 and ban12) were used to generate marked mutant GSC clones for an analysis of ban function in GSCs. The rates of GFP-marked GSCs were measured at two days, seven days, 12 days, and 15 days after heat-shock treatment (AHST). As shown in Figure 5E, compared with FRT control GSC clones, the marked clone rates of both ban20 and ban12 were not reduced significantly during the testing period, indicating that ban is dispensable for GSCs. Together with the phenotypic analyses of ban, our data indicate that, as with dFmr1, ban functions as an extrinsic factor for GSC maintenance (Figure 5).


The bantam microRNA is associated with drosophila fragile X mental retardation protein and regulates the fate of germline stem cells.

Yang Y, Xu S, Xia L, Wang J, Wen S, Jin P, Chen D - PLoS Genet. (2009)

The bantam miRNA plays a non-autonomous role in GSC maintenance.GSC clones were induced by heat-shock treatment in adult female flies. Ovaries from FRT control flies (A and B) and FRT, ban flies (C and D) were dissected at day 2 and day 12 following heat-shock treatment; GSC clones were identified by the lack of GFP expression. Scale bar represents 10 µm. (E) Relative percentages of negatively GFP-marked GSC clones in FRT control and two FRT; ban  alleles at days 2, 7, 12, and 15 AHST are shown. P>0.5.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2654963&req=5

pgen-1000444-g005: The bantam miRNA plays a non-autonomous role in GSC maintenance.GSC clones were induced by heat-shock treatment in adult female flies. Ovaries from FRT control flies (A and B) and FRT, ban flies (C and D) were dissected at day 2 and day 12 following heat-shock treatment; GSC clones were identified by the lack of GFP expression. Scale bar represents 10 µm. (E) Relative percentages of negatively GFP-marked GSC clones in FRT control and two FRT; ban alleles at days 2, 7, 12, and 15 AHST are shown. P>0.5.
Mentions: The loss of germline stem cells in ban mutant ovaries indicates that ban is required by either GSCs or somatic cells. To analyze whether ban functions as a cell-autonomous factor in maintaining GSC fate, we used a FLP-FRT–mediated mitotic recombination technique to generate marked mutant GSCs [17],[22], then calculated the life span of the marked mutant GSCs by quantifying their loss rate. The marked mutant GSCs were identified by a lack of GFP fluorescence in the nuclei and by their positions directly attaching to the base cells of the terminal filament or cap cells. The ban loss-of-function alleles (ban20 and ban12) were used to generate marked mutant GSC clones for an analysis of ban function in GSCs. The rates of GFP-marked GSCs were measured at two days, seven days, 12 days, and 15 days after heat-shock treatment (AHST). As shown in Figure 5E, compared with FRT control GSC clones, the marked clone rates of both ban20 and ban12 were not reduced significantly during the testing period, indicating that ban is dispensable for GSCs. Together with the phenotypic analyses of ban, our data indicate that, as with dFmr1, ban functions as an extrinsic factor for GSC maintenance (Figure 5).

Bottom Line: We show that, like dFmr1, bantam is not only required for repressing primordial germ cell differentiation, it also functions as an extrinsic factor for germline stem cell maintenance.Furthermore, we find that bantam genetically interacts with dFmr1 to regulate the fate of germline stem cells.Collectively, our results support the notion that the FMRP-mediated translation pathway functions through specific miRNAs to control stem cell regulation.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, People's Republic of China.

ABSTRACT
Fragile X syndrome, a common form of inherited mental retardation, is caused by the loss of fragile X mental retardation protein (FMRP). We have previously demonstrated that dFmr1, the Drosophila ortholog of the fragile X mental retardation 1 gene, plays a role in the proper maintenance of germline stem cells in Drosophila ovary; however, the molecular mechanism behind this remains elusive. In this study, we used an immunoprecipitation assay to reveal that specific microRNAs (miRNAs), particularly the bantam miRNA (bantam), are physically associated with dFmrp in ovary. We show that, like dFmr1, bantam is not only required for repressing primordial germ cell differentiation, it also functions as an extrinsic factor for germline stem cell maintenance. Furthermore, we find that bantam genetically interacts with dFmr1 to regulate the fate of germline stem cells. Collectively, our results support the notion that the FMRP-mediated translation pathway functions through specific miRNAs to control stem cell regulation.

Show MeSH
Related in: MedlinePlus