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| Status |
Public on Apr 28, 2021 |
| Title |
RNA profiling of mouse spinal cord ependymal stem cells in vivo and in vitro in different conditions |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by array
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| Summary |
After spinal cord injury, ependymal cells considered as stem cells activate, proliferate and differentiate mainly into glial cells. To understand this further at the molecular level, we performed RNA profiling of these cells in situ using laser-dissection and also when they are cultured as neurospheres in different conditions (growth, differentiation, dedifferentiation)
Abstract: Numerous vertebrates, including Human, maintain a pool of immature cells in the ependymal region of the adult spinal cord. During injury, these ependymal cells, considered as multipotent stem cells, rapidly activate, proliferate and generate neurons and glial cells in lower vertebrates or mainly glial cells in mammals. The mechanisms underlying this activation are ill-defined and we intended to fill this gap by performing RNA profiling of mouse ependymal region after lesion. Bioinformatics and immunofluorescence identified activation of STAT3 and ERK/MAPK signaling in ependymal cells after injury. This was also accompanied by downregulation of cilia-associated genes and FoxJ1, a central transcription factor of ciliogenesis. Six genes were upregulated more than 20 fold, namely Crym, Ecm1, Ifi202b, Nupr1, Osmr, Rbp1, Thbs2 whereas only one, Acta1 was downregulated to this extent. We explored further the role and regulation in ependymal cells of Osmr, the receptor for oncostatin (OSM). This inflammatory cytokine is specifically expressed by microglia cells and we observed interactions between these cells and ependymal cells in vivo. Using culture of ependymal cells in neurospheres, we found that several cytokines induced OSMR, OSM being the most potent. OSMR is also upregulated by co-culture with OSM-expressing microglial cells. Treatment of spinal cord neural stem cells with OSM decreased their proliferation, upregulate p-Stat3 and reduced their differentiation into oligodendrocyte-lineage Olig1+ cells. These results suggest an important role for microglia-derived oncostatin in the activation and fate of spinal cord ependymal cell.
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| Overall design |
We extracted RNA from laser-microdissected ependymal cells in the normal mouse spinal cord and after 3 days post injury (needle insertion). We also extracted RNA from neurospheres derived from the ependymal region and placed in media with and without FGF2 and EGF. RNA profilings were performed using affymetrix microarrays
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| Contributor(s) |
Hugnot J |
| Citation(s) |
34943841 |
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| Submission date |
Apr 30, 2020 |
| Last update date |
Jan 01, 2022 |
| Contact name |
jean-philippe hugnot |
| E-mail(s) |
[email protected]
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| Organization name |
inserm inm u1191
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| Lab |
brain plasticity, stem cell and glial tumors
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| Street address |
IGF INSERM UM 141 rue de la cardonille
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| City |
MOntpellier |
| State/province |
languedoc |
| ZIP/Postal code |
34094 |
| Country |
France |
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| Platforms (2) |
| GPL11180 |
[HT_MG-430_PM] Affymetrix HT MG-430 PM Array Plate |
| GPL17400 |
[MoGene-2_1-st] Affymetrix Mouse Gene 2.1 ST Array [transcript (gene) version] |
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| Samples (24)
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| GSM4508598 |
spinal cord ependymal region, control, mouse 1 |
| GSM4508599 |
spinal cord ependymal region, control, mouse 2 |
| GSM4508600 |
spinal cord ependymal region, control, mouse 3 |
| GSM4508601 |
spinal cord ependymal region, control, mouse 4 |
| GSM4508602 |
spinal cord ependymal region, control, mouse 5 |
| GSM4508603 |
spinal cord ependymal region, control, mouse 6 |
| GSM4508604 |
spinal cord ependymal region, 3 days post injury mouse 1 |
| GSM4508605 |
spinal cord ependymal region, 3 days post injury mouse 2 |
| GSM4508606 |
spinal cord ependymal region, 3 days post injury mouse 3 |
| GSM4508607 |
spinal cord ependymal region, 3 days post injury mouse 4 |
| GSM4508608 |
spinal cord ependymal region, 3 days post injury mouse 5 |
| GSM4508609 |
spinal cord ependymal region, 3 days post injury mouse 6 |
| GSM4508610 |
spinal cord neurospheres in growing condition with EGF and FGF2, mouse 1 |
| GSM4508611 |
spinal cord neurospheres in growing condition with EGF and FGF2, mouse 2 |
| GSM4508612 |
spinal cord neurospheres in growing condition with EGF and FGF2, mouse 3 |
| GSM4508613 |
spinal cord neurospheres in growing condition with EGF and FGF2, mouse 4 |
| GSM4508614 |
spinal cord neurospheres in differentiation condition without EGF and FGF2, mouse 1 |
| GSM4508615 |
spinal cord neurospheres in differentiation condition without EGF and FGF2, mouse 2 |
| GSM4508616 |
spinal cord neurospheres in differentiation condition without EGF and FGF2, mouse 3 |
| GSM4508617 |
spinal cord neurospheres in differentiation condition without EGF and FGF2, mouse 4 |
| GSM4508618 |
spinal cord neurospheres in dedifferentiation condition with EGF and FGF2, mouse 1 |
| GSM4508619 |
spinal cord neurospheres in dedifferentiation condition with EGF and FGF2, mouse 2 |
| GSM4508620 |
spinal cord neurospheres in dedifferentiation condition with EGF and FGF2, mouse 3 |
| GSM4508621 |
spinal cord neurospheres in dedifferentiation condition with EGF and FGF2, mouse 4 |
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| Relations |
| BioProject |
PRJNA629577 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE149669_RAW.tar |
89.9 Mb |
(http)(custom) |
TAR (of CEL, CHP) |
| Processed data included within Sample table |
| Processed data provided as supplementary file |
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