Circulating transforming growth factor-β1 facilitates remyelination in the adult central nervous system
- Osaka University, Japan
- National Center of Neurology and Psychiatry, Japan
- Toneyama National Hospital, Japan
Figures
Figure 1 with 1 supplement
TGF-β1 drives oligodendrocyte maturation.
(A) (Upper panels) Representative images of the oligodendrocyte cultures. Cells were stained with MBP (green) and Olig2 (magenta). (Lower panels) Yellow outlines were obtained from HCA to measure …
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Figure 1—source data 1
Pharmacological impact for serum-mediated oligodendrocyte maturation.
Inhibitors were used at a final concentration of 10 μM. Each line shows the relative values of MBP/Olig2 area (two rows from the left) and Olig2 cell number (two rows from the right) relative to the control value (DMSO treatment without serum). TGF-βRI kinase inhibitor (red letters) is the only drug we tested that targets a receptor-type protein. The data is average of two trials.
- https://doi.org/10.7554/eLife.41869.004
Figure 1—figure supplement 1
A heat-resistant protein in circulation promotes oligodendrocyte maturation.
(A) Representative images of the oligodendrocyte culture. Serum was pretreated with the indicated reagent and added into the culture. Cells were cultured 5 days after treatment. (B) Quantification …
Figure 2
TGF-β1 level in circulation is higher than that in the CNS.
(A) Quantification of TGF-β1 protein level in individual organs in intact mice (n = 3 for each), p<0.0001. (B) Quantification of TGF-β1 protein concentration in the serum and CSF (n = 3 for serum, n …
Figure 3 with 1 supplement
Circulating TGF-β1 supports spontaneous remyelination.
(A) Quantification of circulating TGF-β1 level in the mice with platelet depletion. Plasma from mice were obtained from 7 days after the start of anti-CD41 mAbs injection (n = 7 for control IgG, n = …
Figure 3—figure supplement 1
Circulating TGF-β leaks into the spinal cord after LPC injection.
(A) Quantification of TGF-β1 level in the spleen in mice subjected to platelet depletion. Samples were collected 7 days after the start of anti-CD41 mAb injection (n = 4 for each), p=0.0129. (B) …
Figure 4 with 1 supplement
TGF-βRI in oligodendrocytes is required for remyelination.
(A) Representative images of spinal cord sections double-labeled for APC and TGF-βRI. Spinal cords were obtained from control mice. The white arrowheads show APC+ TGF-βRI+ cells. (B) Representative …
Figure 4—figure supplement 1
TGF-βRI is expressed in oligodendrocytes.
(A) Representative images of spinal cord sections double-labeled for PDGFRα (magenta) and with TGF-βRI (green). (B) Representative images of spinal cord sections double-labeled for APC (magenta) and …
Figure 5 with 1 supplement
TGF-β1 treatment promotes remyelination.
(A) Representative image of brain section labeled for MBP. Brains were obtained from the cuprizone fed mice, along with TGF-β1 administration for 14 days after removal of cuprizone diet. (B) …
Figure 5—figure supplement 1
TGF-β treatment does not regulate accumulation of inflammatory cells.
(A) Representative images of spinal cord section stained with hematoxylin and eosin. Spinal cord section was collected 14 days after the start of TGF-β1 treatment. Right panels show high …
Figure 6
TGF-β1 stimulates human oligodendrocyte maturation.
(A) Representative images of spinal cord sections double-labeled for APC with TGF-βRI. Brain samples were obtained post-mortem from MS patients. (B) Relative expression of indicated mRNA in human …
Author response image 1
Tables
Key resources table
| Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|
| Tg(Plp1-cre/ERT)3Pop | PMID:12481300 | RRID:MGI:3696409 | |
| ALK5 flox | PMID:12842983 | Dr. Stefan Karlsson (Lund University) | |
| Myelin Basic Protein antibody | PMID:20209960 | RRID:AB_305869 | IHC (1:500) |
| Goat Anti-Human Olig2 Polyclonal antibody | PMID:18615534 | RRID:AB_2157554 | IHC (1:300) |
| Anti-APC (Ab-7) Mouse mAb (CC-1) antibody | PMID:16705673 | RRID:AB_2057371 | IHC (1:500) |
| Donkey Anti-Mouse IgG (H + L) Antibody, Alexa Fluor 594 Conjugated | PMID:23970788 | RRID:AB_141633 | IHC (1:500) |
| Donkey Anti-Goat IgG (H + L) Antibody , Alexa Fluor 647 Conjugated | PMID:25505326 | RRID:AB_141844 | IHC (1:500) |
| Phospho-Smad2 (Ser465/467)/Smad3 (Ser423/425) (D27F4) Rabbit mAb antibody | PMID:28324027 | RRID:AB_2631089 | WB (1:1000) |
| Smad2/3 (D7G7) XP Rabbit mAb antibody | PMID:29056512 | RRID:AB_10889933 | WB (1:1000) |
| Anti-Rabbit IgG Phototope-HRP Western Blot Detection System Antibody, Horseradish Peroxidase Conjugated | RRID:AB_2099234 | WB (1:2000) | |
| Rat Anti-CD41 Monoclonal Antibody, Unconjugated, Clone MWReg30 | PMID:30590031 | RRID:AB_395084 | |
| Goat Anti-Mouse Pdgf r alpha Polyclonal antibody, Unconjugated | PMID:28111199 | RRID:AB_2236897 | IHC (1:500) |
| TGFbeta RI (V-22) antibody | PMID:29161592 | RRID:AB_632493 | IHC (1:50) |
| Donkey Anti-Rat IgG (H + L) Antibody, Alexa Fluor 488 Conjugated | PMID:28280459 | RRID:AB_141709 | IHC (1:500) |
| TGF-beta 1,–2,−3 MAb (Clone 1D11) antibody | PMID: 28521131 | RRID:AB_357931 | |
| Q-Plex Mouse Cytokine - Screen (16-Plex) | Quansys Biosciences | Cat. #110949 MS | |
| Human Oligodendrocyte differentiation Kit | Millipore | Cat. #CS204496 | |
| Human recombinant TGF-β1 | R and D Systems | Cat. #240-B | |
| Recombinant Murine basic FGF | Peprotech | Cat. #450–33 | |
| Recombinant Murine PDGF-AA | Peprotech | Cat. #315–17 | |
| LY364947 | Calbiochem | Cat. #616451 | |
| Recombinant mouse TGF-b1 | R and D Systems | Cat. #7666 MB-005 | |
| Mouse/Rat/Porcine/Canine TGFb1 Quantikine ELISA | R and D Systems | Cat. #MB100B | |
| Cell Proliferation ELISA, BrdU (colorimetric) | Sigma Aldrich | Cat. #11647229001 | |
| L-α-Lysophosphatidylcholine (LPC) from bovine brain | Sigma Aldrich | Cat. #L1381 | |
| InhibitorSelect 384-Well Protein Kinase Inhibitor Library I | Calbiochem | Cat. #539743 |
Additional files
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Transparent reporting form
- https://doi.org/10.7554/eLife.41869.013
