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GEO help: Mouse over screen elements for information. |
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| Status |
Public on Jan 12, 2018 |
| Title |
Troy+ brain stem cells cycle through quiescence and regulate their number by sensing niche occupancy |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
The adult mouse subependymal zone (SEZ) provides a niche for mammalian neural stem cells (NSCs). However, the molecular signature, self-renewal potential and fate behavior of NSCs remain poorly defined. Here we propose a model in which the fate of active NSCs is coupled to the total number of neighboring NSCs in a shared niche. Using knock-in reporter alleles and single-cell RNA sequencing, we show that the Wnt target Tnfrsf19/Troy identifies both active and quiescent NSCs. Quantitative analysis of genetic lineage tracing of individual NSCs under homeostasis or in response to injury reveals rapid expansion of stem cell number before some return to quiescence. This behavior is best explained by stochastic fate decisions, where stem cell number within a shared niche fluctuates over time. Fate-mapping proliferating cells using a novel Ki67iresCreER allele confirms that active NSCs reversibly return to quiescence, achieving long-term self-renewal. Our findings suggest a novel niche-based mechanism for the regulation of NSC fate and number.
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| Overall design |
We analysed the adult mouse subependymal zone (SEZ), the largest neruogenic niche in the brain, at a single cell level. For this, we used SORT-seq, a combination of FACS and single cell RNA-sequencing that resulted in high quality transcriptome data from ~1500 single cells (out of ~2500 cells sequenced). To identify different cell types, we used knock-in mouse models, surface labeling using antibdoies as well as lineage tracing. Troy-GFPiresCreER +/ki mice were used to identify NSCs. Ki67-tagRFP ki/ki mice were used to identify dividing cells. Ki67-iresCreER +/ki Rosa-tdTomato +/ki mice were administered a single injection of 5mg of Tamoxifen to label dividing cells (day2 - d2) and follow their progeny (day60 - d60) in the SEZ as well as the olfactory bulb (for d60 only). We also included published markers; combinations of anti-GLAST antibody (to identify NSCs) and EGF binding ability using alexa647-conjugated EGF (to identify dividing cells), combinations of anti-Prom1 (Cell contacting the ventricles) and anti-CD24 antibodies (high in neuroblasts), as well as anti-O4 antibody (oligodendrocytes) to create an atlas of NSCs.
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| Contributor(s) |
Basak O, Krieger TG, Muraro MJ, Wiebrands K, Stange DE, Aldeguer JF, Rivron NC, van de Wetering M, van Es JH, van Oudenaarden A, Simons BD, Clevers H |
| Citation(s) |
29311336 |
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| Submission date |
Feb 17, 2015 |
| Last update date |
May 31, 2019 |
| Contact name |
Onur BASAK |
| E-mail(s) |
onurbasak@yahoo.com
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| Organization name |
Hubrecht Institute
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| Lab |
Clevers
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| Street address |
Uppsalalaan 8
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| City |
Utrecht |
| ZIP/Postal code |
3584CT |
| Country |
Netherlands |
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| Platforms (1) |
| GPL19057 |
Illumina NextSeq 500 (Mus musculus) |
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| Samples (28)
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| Relations |
| BioProject |
PRJNA275609 |
| SRA |
SRP055081 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE65970_RAW.tar |
9.1 Mb |
(http)(custom) |
TAR (of CSV) |
| GSE65970_cel-seq_barcodes.csv.gz |
424 b |
(ftp)(http) |
CSV |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
| Processed data provided as supplementary file |
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