TEAD1 is crucial for developmental myelination, Remak bundles, and functional regeneration of peripheral nerves

  1. Matthew Grove
  2. Hyukmin Kim
  3. Shuhuan Pang
  4. Jose Paz Amaya
  5. Guoqing Hu
  6. Jiliang Zhou
  7. Michel Lemay
  8. Young-Jin Son  Is a corresponding author
  1. Department of Neural Sciences, Shriners Hospitals Pediatric Research Center, Lewis Katz School of Medicine, Temple University, United States
  2. Department of Bioengineering, Temple University, United States
  3. Department of Pharmacology & Toxicology, Medical College of Georgia, Augusta University, United States
8 figures, 1 video, 1 table and 1 additional file

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—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 …

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 …

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
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 …

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
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 …

Videos

Video 1
A movie showing a P55 WT and two littermates Tead1 cKO mic.

Tables

Key resources table
Reagent type (species) or resourceDesignationSource or referenceIdentifiersAdditional information
Strain, strain background (Mus musculus)C57Bl/6Jackson LaboratoryStock #: 000664; RRID:IMSR JAX:000664
Genetic reagent (M. musculus)Plp1-Cre-ERT2Leone et al., 2003MGI:2663093
Antibodyanti-Yap/Taz (rabbit monoclonal)Cell Signaling TechnologyD24E4, #8418 RRID:AB_10950494IHC 1:200 Western 1:1000
Antibodyanti-SCG10 (rabbit monoclonal)Novus BiologicalsNBP1-49461 RRID:AB_10011569IHC 1:5000
Antibodyanti-Yap (rabbit monoclonal)Cell Signaling TechnologyD8H1X, #14074 RRID:AB_2650491IHC 1:200
Antibodyanti-Sox10 (goat polyclonal)R&D Systems#AF-2864 RRID:AB_442208IHC 1:100
Antibodyanti-Sox10 (rabbit monoclonal)AbcamEPR4007, #ab155279 RRID:AB_2650603IHC 1:250
Antibodyanti-Egr2 (rabbit polyclonal)Professor Dies Meijer, University of EdinburghIHC 1:4000
Antibodyanti-Oct6 (rabbit monoclonal)AbcamEP5421, #ab126746 RRID:AB_11130256WB 1:1000
AntibodyAnti-Oct6 (rabbit polyclonal)Abcam#ab31766 RRID:AB_776899IHC 1:800
AntibodyAnti-c-Jun (mouse monoclonal)BD Transduction Laboratories#610326 RRID:AB_397716IHC 1:500
AntibodyAnti-c-Jun (rabbit monoclonal)Cell Signaling Technology60 A8, #9165 RRID:AB_2130165WB 1:1000
AntibodyAnti-pS63-c-Jun (rabbit polyclonal)Cell Signaling Technology#9261 RRID:AB_2130162IHC 1:100
Antibodyanti-Ki67 (rabbit polyclonal)Abcam#ab15580 RRID:AB_443209IHC 1:200
Antibodyanti-p75NGFR (goat polyclonal)Neuromics#GT15057 RRID:AB_2737189IHC 1:400
Antibodyanti-Tubulin β3 (rabbit polyclonal)Biolegend#802001 RRID:AB_2564645IHC 1:1000
AntibodyIRDye-680 (goat anti-mouse)LI-COR#926–32220 RRID:AB_621840WB 1:15,000
AntibodyHRP-Goat anti-mouse secondary antibodyJackson Immunoresearch#715-035-150 RRID:AB_2340770WB 1:12,000
AntibodyHRP-Goat anti-rabbit secondary antibodyJackson Immunoresearch#115-055-062 RRID:AB_2338533WB 1:12,000
Chemical compound, drugAraldite 6005EMS#10920
Chemical compound, drugDDSAEMS#13710
Chemical compound, drugDBPEMS#13101
Chemical compound, drugBDMAEMS#11400–25
OtherCoated grids (100 mesh)EMS#FF100-Cu
Chemical compound, drugOsmium tetroxide (4% solution)EMS#19170
Chemical compound, drugLead nitrateEMS#17900
Chemical compound, drugSodium citrateEMS#21140
Chemical compound, drugUranyl acetateEMS#22400
Chemical compound, drugSodium borateEMS#21130
Chemical compound, drugToluidine blueEMS#22050
Chemical compound, drugParaformaldehydeSigma-Aldrich#158127
Commercial assay or kitClick-It EdU Alexa Fluor 594 kitThermoFisher Scientific#C10339
Chemical compound, drugEdUThermoFisher Scientific#E10187
Chemical compound, drugTamoxifenSigma-Aldrich#T5648
OtherDAPI stainInvitrogen#D1306IHC 1:250
AntibodyAlexa 488, 568 or 647 secondariesJackson ImmunoresearchIHC 1:250 to 1:1000
Software, algorithmImage Studio LiteLI-COR, Inc
Software, algorithmPrismGraphPad Software, Inc
Software, algorithmStataStataCorp LPMann-Whitney test

Additional files

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