Suppressor of fused controls perinatal expansion and quiescence of future dentate adult neural stem cells
- University of California, San Francisco, United States
- Kyushu University, Japan
Figures
Figure 1 with 1 supplement
Deletion of Sufu decreases Hh-responding cells during DG development.
(A) Representative Gli1-LacZ staining images of sagittal brain sections in Sufufl/fl;Gli1lacZ/+and hGFAP-Sufu-KO;Gli1lacZ/+ mice at E15.5-P0. Magnified images of the black dashed-line boxes are …
Figure 1—figure supplement 1
Deletion of Sufu decreases Hh-responding cells during DG development.
Representative Gli1-LacZ staining images of dorsal DG in sagittal sections of Sufufl/fl;Gli1lacZ/+ and hGFAP-Sufu-KO;Gli1lacZ/+ mice. Note that lacZ +cells are observed more in dorsal DG compared …
Figure 2 with 1 supplement
Deletion of Sufu decreases proliferation of DG-NSCs.
(A) Representative immunofluorescence images of Sox2 (red) and Ki67 (green) in the DG of Sufufl/fl and hGFAP-Sufu-KO mice at P7. DNA is stained with DAPI (blue). Magnified images of the white …
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Figure 2—source data 1
Extended numerical data and statistical analysis for Figure 2—figure supplement 1.
- https://doi.org/10.7554/eLife.42918.006
Figure 2—figure supplement 1
Excessive Shh signaling activity leads to abnormal morphology of DGs and increased cell death.
(A) Representative immunofluorescence images of Cleaved Caspase 3 (red) and DAPI (blue) in the DG of WT and hGFAP-Cre;SmoM2 mice at P7. (B, C) Quantification of Cleaved Caspase 3 + cells in the DGs …
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Figure 2—figure supplement 1—source data 1
Extended numerical data and statistical analysis for Figure 2.
- https://doi.org/10.7554/eLife.42918.007
Figure 3 with 2 supplements
Gli1 deletion increases the developmental defects in hGFAP-Cre Sufufl/fl mice.
(A) Representative immunofluorescence images of Sox2 (red), Ki67 (green) and DAPI (blue) in the DG of Control, hGFAP-Sufu-KO, hGFAP-Sufu-KO;Gli1lacZ/+, and hGFAP-Sufu-KO;Gli1lacZ/lacZ mice at P7. (B,…
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Figure 3—source data 1
Extended numerical data and statistical analysis for Figure 3.
- https://doi.org/10.7554/eLife.42918.011
Figure 3—figure supplement 1
Deletion of Gli1 does not affect cell proliferation.
(A) Representative immunofluorescence images of Sox2 (red) and Ki67 (green) in the DG of Sufufl/fl, Sufufl/fl;Gli1lacZ/+, Sufufl/fl;Gli1lacZ/lacZ mice at P7. DNA is stained with DAPI (blue). (B,C) …
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Figure 3—figure supplement 1—source data 1
Extended numerical data and statistical analysis for Figure 3—figure supplement 1.
- https://doi.org/10.7554/eLife.42918.012
Figure 3—figure supplement 2
Deletion of Sufu decreases proliferation of NSCs, but not neuronal precursors.
(A) Representative immunofluorescence images of Sox2 (red) and Ki67 (green) in the DG of Control, hGFAP-Sufu-KO and hGFAP-Sufu-KO;Gli1lacZ/+ mice at P14. DNA is stained with DAPI (blue). (B,C) …
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Figure 3—figure supplement 2—source data 1
Extended numerical data and statistical analysis for Figure 3—figure supplement 2.
- https://doi.org/10.7554/eLife.42918.013
Figure 4 with 1 supplement
Deletion of Sufu during DG development decreases qNSCs pool in adult DGs.
(A) Experimental scheme of BrdU injection. 8 week-old mice were injected with BrdU for 5 days and analyzed 3 days after last BrdU injection. (B) Representative immunofluorescence images for DCX …
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Figure 4—source data 1
Extended numerical data and statistical analysis for Figure 4.
- https://doi.org/10.7554/eLife.42918.016
Figure 4—figure supplement 1
Deletion of Sufu during DG development decreases qNSCs pool in adult DGs.
(A) Representative immunofluorescence images for Tbr2 (red) and DAPI (blue) in the DGs of control, hGFAP-Sufu-KO and hGFAP-Sufu-KO;Gli1lacZ/+ mice. (B) Quantification of Tbr2 +cells in SGZ [Control, …
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Figure 4—figure supplement 1—source data 1
Extended numerical data and statistical analysis for Figure 4—figure supplement 1.
- https://doi.org/10.7554/eLife.42918.017
Figure 5 with 1 supplement
Long-lived NSCs expand in first postnatal week, and loss of Sufu impairs its expansion.
(A) Fate tracing of Hh-responding cells at E17.5 in postnatal DG development. Gli1CreERT2/+ Ai14 mice were treated with tamoxifen at E17.5 and Ai14 +Hh responding cells are analyzed in postnatal …
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Figure 5—source data 1
Extended numerical data and statistical analysis for Figure 5.
- https://doi.org/10.7554/eLife.42918.020
Figure 5—figure supplement 1
Prenatal deletion of Sufu in NSCs of ventral hippocampus leads to reduction of NSCs in the postnatal DGs.
(A) Representative immunofluorescence images for Ai14 (red), Sox2 (green) and DAPI (blue) in the DGs of Gli1CreERT2/+;Sufufl/+;Ai14 and Gli1CreERT2/+;Sufufl/fl;Ai14 mice. (B,C) Quantification of …
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Figure 5—figure supplement 1—source data 1
Extended numerical data and statistical analysis for Figure 5—figure supplement 1.
- https://doi.org/10.7554/eLife.42918.021
Figure 6 with 2 supplements
Deletion of Sufu prematurely induces the quiescent state transition.
(A) Schematic illustration for labeling NSCs that established quiescent state during DG development using two thymidine analogs. CldU is diluted when cell divides. Thus CldU amount goes down if the …
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Figure 6—source data 1
Extended numerical data and statistical analysis for Figure 6.
- https://doi.org/10.7554/eLife.42918.025
Figure 6—figure supplement 1
Deletion of Sufu prematurely induces the quiescent state transition.
(A) Representative immunofluorescence images for CldU (green), Sox2 (red), GFAP (cyan) and DAPI (blue) in the SGZ of animals injected CldU at different stages. White arrowheads indicate the …
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Figure 6—figure supplement 1—source data 1
Extended numerical data and statistical analysis for Figure 6—figure supplement 1.
- https://doi.org/10.7554/eLife.42918.026
Figure 6—figure supplement 2
Deletion of Sufu prematurely decreases the proliferation of NSCs during DG development.
(A) Representative immunofluorescence images of Sox2 (red) and Ki67 (green) in the dorsal and ventral DG of control, hGFAP-Sufu-KO and hGFAP-Sufu-KO;Gli1lacZ/+ mice at P0. DNA is stained with DAPI …
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Figure 6—figure supplement 2—source data 1
Extended numerical data and statistical analysis for Figure 6—figure supplement 2.
- https://doi.org/10.7554/eLife.42918.027
Figure 7
Sufu is important for the perinatal expansion and quiescent state transition of dentate NSCs.
Schematic summary illustrating the role of Sufu and Shh signaling activity in initial production, expansion and quiescent state transition of NSCs during DG development. Long-lived NSCs are produced …
Author response image 1
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Genetic reagent (M. musculus) | Sufuflox/flox | PMID: 20074523 | RRID: MGI:4840420 | Dr. Chi-Chung Hui (University of Toronto) |
| Genetic reagent (M. musculus) | hGFAP-Cre | PMID: 11668683 | RRID: MGI:2179048 | Jackson Laboratory (Stock:004600) |
| Genetic reagent (M. musculus) | Gli1CreERT2/+ | PMID: 15315762 | RRID: MGI:3053957 | Jackson Laboratory (Stock:007913) |
| Genetic reagent (M. musculus) | Gli1LacZ/+ | PMID: 12361967 | RRID: MGI:J:79392 | Jackson Laboratory (Stock:008211) |
| Genetic reagent (M. musculus) | Rosa-Ai14 | PMID: 20023653 | RRID: MGI:J:155793 | Jackson Laboratory (Stock:007908) |
| Genetic reagent (M. musculus) | SmoM2-YFP | PMID: 15107405 | RRID: MGI:3576373 | Jackson Laboratory (Stock:005130) |
| Antibody | Mouse monoclonal anti-Ki67 | BD Biosciences | RRID: AB_396287 | IHC (1:500) |
| Antibody | rabbit monoclonal anti-Sox2 | Abcam | RRID: AB_10585428 | IHC (1:250) |
| Antibody | Rabbit polyclonal anti-Tbr2 | Abcom | RRID: AB_778267 | IHC(1:250) |
| Antibody | rabbit polyclonal anti-DCX | Abcam | RRID: AB_732011 | IHC (1:1000) |
| Antibody | rat monoclonal anti-GFAP | Zymed | RRID: AB_2532994 | IHC (1:500) |
| Antibody | chickin polyclonal anti-GFAP | Millipore | RRID: AB_177521 | IHC (1:1000) |
| Antibody | rat monoclonal anti-RFP | Chromotek | RRID: AB_2336064 | IHC (1:1000) |
| Antibody | rat monoclonal anti-BrdU | Abcam | RRID: AB_305426 | IHC (1:500) CldU detection |
| Antibody | mouse monoclonalanti-BrdU | BD Biosciences | RRID: AB_400326 | IHC (1:100) IdU detection |
| Antibody | rabbit monoclonalaniti-Cleaved Caspase 3 | Cell Signaling | RRID: AB_2070042 | IHC (1:250) |
| Software | ImageJ | NIH | RRID: SCR_003070 | Cell counting |
| Software | Prism 7 | Graphpad | RRID: SCR_002798 | Statistuc analysis |
Additional files
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Supplementary file 1
The primers for qPCR analysis.
- https://doi.org/10.7554/eLife.42918.029
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Transparent reporting form
- https://doi.org/10.7554/eLife.42918.030
