TEAD1 is crucial for developmental myelination, Remak bundles, and functional regeneration of peripheral nerves
- Department of Neural Sciences, Shriners Hospitals Pediatric Research Center, Lewis Katz School of Medicine, Temple University, United States
- Department of Bioengineering, Temple University, United States
- Department of Pharmacology & Toxicology, Medical College of Georgia, Augusta University, United States
Figures
Figure 1 with 1 supplement
Multiple TEADs are expressed in Schwann cells of WT and Tead1 cKO mice.
(A) Immunostaining and quantification of TEAD1 +SCs on longitudinal sections of P22 WT and Tead1 cKO SNs. SC nuclei are identified by Sox10. All cell nuclei are marked by DAPI. n=3 mice per …
-
Figure 1—source data 1
This zip archive contains source files for graphs in Figure 1A, B, C and D and uncropped labeled or unlabeled blots of Figure 1B and D.
- https://cdn.elifesciences.org/articles/87394/elife-87394-fig1-data1-v2.zip
Figure 1—figure supplement 1
Western blotting of P5, P38, P85 WT, and P38 Tead1 cKO SN lysates with anti-TEAD2.
TEAD2 expression is normalized to that of β-actin as an internal control. Arrow denotes ~65 kD TEAD2 band that was diminished with age.
-
Figure 1—figure supplement 1—source data 1
This zip archive contains uncropped labeled or unlabeled blots in Figure 1—figure supplement 1.
- https://cdn.elifesciences.org/articles/87394/elife-87394-fig1-figsupp1-data1-v2.zip
Figure 2 with 1 supplement
TEAD1 is required for myelin sheath formation and growth.
(A) A living control (WT) and two Tead1 cKO mice from one litter at P55. Arrows point to splayed paralyzed hindlimbs. (B) Exposed SNs of WT and Tead1 cKO mice during dissection. Scale bar: 1 mm (C) …
-
Figure 2—source data 1
This zip archive contains source files for graphs in Figure 2D, H, I, J, K and M.
- https://cdn.elifesciences.org/articles/87394/elife-87394-fig2-data1-v2.zip
Figure 2—figure supplement 1
Additional semi-thin and TEM images of WT and Tead1 cKO.
(A) Representative images of semi-thin sections of various areas of WT and Tead1 cKO SNs at P22 and P50. Scale bar: 20 μm (B) Representative TEM images of transverse sections of WT and Tead1 cKO SNs …
Figure 3
TEAD1 regulates SC proliferation both positively and negatively.
(A) Immunostaining and quantification of SCs on transverse sections of WT and Tead1 cKO SNs at P3, P22, and P52. SC nuclei and all cell nuclei are marked by Sox10 and DAPI. Axons are marked by …
-
Figure 3—source data 1
This zip archive contains source files for graphs in Figure 3A, B, C and D.
- https://cdn.elifesciences.org/articles/87394/elife-87394-fig3-data1-v2.zip
Figure 4
TEAD1 is required for Krox20 to upregulate myelin proteins during development.
(A) Western blotting of P50 WT and Tead1 cKO SN lysates with anti-Oct6, -Krox20, -MPZ, and -MBP. Protein expression is normalized to that of β-actin as an internal control, and WT expression is …
-
Figure 4—source data 1
This zip archive contains source files for graphs in Figure 4A, B, C and D and uncropped labeled or unlabeled blots of Figure 4A.
- https://cdn.elifesciences.org/articles/87394/elife-87394-fig4-data1-v2.zip
Figure 5
TEAD1 and YAP/TAZ upregulate cholesterol synthesis enzymes: FDPS and IDI1.
(A) Western blotting of P8 and P40 WT and Tead1 cKO SN lysates with anti-SREBP1, -SCD1, -SREBP2, -HMGCR, -FDPS, and -IDI1. Protein expression is normalized to that of β-actin as an internal control, …
-
Figure 5—source data 1
This zip archive contains uncropped labeled or unlabeled blots of P8 WT and Tead1 cKO SNs in Figure 5A.
- https://cdn.elifesciences.org/articles/87394/elife-87394-fig5-data1-v2.zip
-
Figure 5—source data 2
This zip archive contains source files for graphs in Figure 5A and B, and uncropped labeled or unlabeled blots of P40 WT and Tead1 cKO SNs in Figure 5A and P60 SNs of WT and YAP/TAZ cDKO in Figure 5B.
- https://cdn.elifesciences.org/articles/87394/elife-87394-fig5-data2-v2.zip
Figure 6 with 1 supplement
TEAD1 is largely dispensable for myelin maintenance, but required for Remak bundle integrity.
(A) Cartoon showing timeline of tamoxifen (TAM) injection and time of sacrifice of Tead1 iKO mice. (B) Representative TEM images of P150 WT and Tead1 iKO SNs showing occasional large axons …
-
Figure 6—source data 1
This zip archive contains source files for graphs in Figure 6C and E.
- https://cdn.elifesciences.org/articles/87394/elife-87394-fig6-data1-v2.zip
-
Figure 6—source data 2
This zip archive contains source files for graphs in Figure 6A–G and uncropped labeled or unlabeled blots of Figure 6A.
- https://cdn.elifesciences.org/articles/87394/elife-87394-fig6-data2-v2.zip
Figure 6—figure supplement 1
Expression of TEAD1, pan-TEAD and YAP/TAZ in intact and regenerating nerves of WT and Tead1 iKO mice.
(A) Western blotting of adult WT and Tead1 iKO SN lysates with anti-TEAD1. TEAD1 expression is normalized to that of β-actin as an internal control, and WT expression is arbitrarily given the value …
Figure 7
TEAD1 is dispensable for YAP/TAZ to maintain Krox20, FDP and IDI1 expression.
(A) Immunostaining and quantification of Krox20 +SCs on longitudinal sections of WT and Yap1/Wwtr1 iDKO SNs 2 weeks after 1st tamoxifen injection. SC nuclei are marked by Sox10. All cell nuclei are …
-
Figure 7—source data 1
This zip archive contains source files for graphs in Figure 7A–E and uncropped labeled or unlabeled blots of Figure 7D and E.
- https://cdn.elifesciences.org/articles/87394/elife-87394-fig7-data1-v2.zip
Figure 8
TEAD1 is required for functional regeneration of peripheral nerve.
(A) Cartoon showing timeline of tamoxifen (TAM) injection and time of sacrifice of Tead1 iKO mice after nerve crush. (B) Low magnification images of longitudinal sections of ~4 mm long SN segments …
-
Figure 8—source data 1
This zip archive contains source files for graphs in Figure 8E, F, G and H.
- https://cdn.elifesciences.org/articles/87394/elife-87394-fig8-data1-v2.zip
Videos
Video 1
A movie showing a P55 WT and two littermates Tead1 cKO mic.
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Mus musculus) | C57Bl/6 | Jackson Laboratory | Stock #: 000664; RRID:IMSR JAX:000664 | |
| Genetic reagent (M. musculus) | Plp1-Cre-ERT2 | Leone et al., 2003 | MGI:2663093 | |
| Antibody | anti-Yap/Taz (rabbit monoclonal) | Cell Signaling Technology | D24E4, #8418 RRID:AB_10950494 | IHC 1:200 Western 1:1000 |
| Antibody | anti-SCG10 (rabbit monoclonal) | Novus Biologicals | NBP1-49461 RRID:AB_10011569 | IHC 1:5000 |
| Antibody | anti-Yap (rabbit monoclonal) | Cell Signaling Technology | D8H1X, #14074 RRID:AB_2650491 | IHC 1:200 |
| Antibody | anti-Sox10 (goat polyclonal) | R&D Systems | #AF-2864 RRID:AB_442208 | IHC 1:100 |
| Antibody | anti-Sox10 (rabbit monoclonal) | Abcam | EPR4007, #ab155279 RRID:AB_2650603 | IHC 1:250 |
| Antibody | anti-Egr2 (rabbit polyclonal) | Professor Dies Meijer, University of Edinburgh | IHC 1:4000 | |
| Antibody | anti-Oct6 (rabbit monoclonal) | Abcam | EP5421, #ab126746 RRID:AB_11130256 | WB 1:1000 |
| Antibody | Anti-Oct6 (rabbit polyclonal) | Abcam | #ab31766 RRID:AB_776899 | IHC 1:800 |
| Antibody | Anti-c-Jun (mouse monoclonal) | BD Transduction Laboratories | #610326 RRID:AB_397716 | IHC 1:500 |
| Antibody | Anti-c-Jun (rabbit monoclonal) | Cell Signaling Technology | 60 A8, #9165 RRID:AB_2130165 | WB 1:1000 |
| Antibody | Anti-pS63-c-Jun (rabbit polyclonal) | Cell Signaling Technology | #9261 RRID:AB_2130162 | IHC 1:100 |
| Antibody | anti-Ki67 (rabbit polyclonal) | Abcam | #ab15580 RRID:AB_443209 | IHC 1:200 |
| Antibody | anti-p75NGFR (goat polyclonal) | Neuromics | #GT15057 RRID:AB_2737189 | IHC 1:400 |
| Antibody | anti-Tubulin β3 (rabbit polyclonal) | Biolegend | #802001 RRID:AB_2564645 | IHC 1:1000 |
| Antibody | IRDye-680 (goat anti-mouse) | LI-COR | #926–32220 RRID:AB_621840 | WB 1:15,000 |
| Antibody | HRP-Goat anti-mouse secondary antibody | Jackson Immunoresearch | #715-035-150 RRID:AB_2340770 | WB 1:12,000 |
| Antibody | HRP-Goat anti-rabbit secondary antibody | Jackson Immunoresearch | #115-055-062 RRID:AB_2338533 | WB 1:12,000 |
| Chemical compound, drug | Araldite 6005 | EMS | #10920 | |
| Chemical compound, drug | DDSA | EMS | #13710 | |
| Chemical compound, drug | DBP | EMS | #13101 | |
| Chemical compound, drug | BDMA | EMS | #11400–25 | |
| Other | Coated grids (100 mesh) | EMS | #FF100-Cu | |
| Chemical compound, drug | Osmium tetroxide (4% solution) | EMS | #19170 | |
| Chemical compound, drug | Lead nitrate | EMS | #17900 | |
| Chemical compound, drug | Sodium citrate | EMS | #21140 | |
| Chemical compound, drug | Uranyl acetate | EMS | #22400 | |
| Chemical compound, drug | Sodium borate | EMS | #21130 | |
| Chemical compound, drug | Toluidine blue | EMS | #22050 | |
| Chemical compound, drug | Paraformaldehyde | Sigma-Aldrich | #158127 | |
| Commercial assay or kit | Click-It EdU Alexa Fluor 594 kit | ThermoFisher Scientific | #C10339 | |
| Chemical compound, drug | EdU | ThermoFisher Scientific | #E10187 | |
| Chemical compound, drug | Tamoxifen | Sigma-Aldrich | #T5648 | |
| Other | DAPI stain | Invitrogen | #D1306 | IHC 1:250 |
| Antibody | Alexa 488, 568 or 647 secondaries | Jackson Immunoresearch | IHC 1:250 to 1:1000 | |
| Software, algorithm | Image Studio Lite | LI-COR, Inc | ||
| Software, algorithm | Prism | GraphPad Software, Inc | ||
| Software, algorithm | Stata | StataCorp LP | Mann-Whitney test |
