YAP/TAZ initiate and maintain Schwann cell myelination
- Lewis Katz School of Medicine, Temple University, United States
- Temple University, United States
Figures
Figure 1 with 1 supplement
YAP/TAZ are largely nuclear in proliferating, differentiating and mature myelinating SCs.
YAP/TAZ localization was investigated in sciatic nerve cryosections (A, B) or teased fibers (C, D). SC nuclei are marked by Sox10 (red) or DAPI (blue). (A) YAP/TAZ (green) are in SC nuclei …
Figure 1—figure supplement 1
Selection of specific antibodies for immunohistochemical analysis of YAP and TAZ.
(A) Western blotting of WT P23 sciatic nerve lysates, showing three different antibodies specifically binding to both YAP/TAZ, only YAP, and only to phosphorylated YAP. (B–D) These antibodies were …
Figure 2 with 3 supplements
YAP/TAZ are required for myelination of sorted axons.
(A) Living control and Yap/Taz DKO mice, showing splayed paralyzed hindlimbs (arrows) and muscle wasting of P60 Yap/Taz cDKO mice. (B) Control sciatic nerve is white and opaque, while cDKO sciatic …
Figure 2—figure supplement 1
Recombination efficiency in Yap cKO, Taz cKO and Yap/Taz cDKO.
(A) Western blotting of sciatic nerve lysates from P60 mice of the indicated genotypes, showing efficient, selective deletion of YAP in Yap cKO, TAZ in Taz cKO, and both YAP/TAZ in Yap/Taz cDKO. …
Figure 2—figure supplement 2
Behavioral tests of motor and sensory function of adult Yap/Taz cDKO mice.
Behavioral tests were conducted on WT or age-matched Yap/Taz cDKO mice. (A) Foot printing showing uncoordinated gait of 10 week- and 16 week-old cDKO mice. (B) Grip strength test, showing markedly …
Figure 2—figure supplement 3
Complete amyelination of forelimb peripheral nerves and nerve roots in P60 Yap/Taz cDKO.
Semi-thin sections of sciatic nerves of WT, Yap cKO and Taz cKO and of forelimb nerve, dorsal root and ventral root of cDKO were stained with toluidine blue. Insets in pink boxes are enlarged areas …
Figure 3
YAP/TAZ are required for timely axon sorting.
(A) Left nine panels: Semi-thin transverse sections of proximal control sciatic nerve and proximal and distal cDKO nerves at P4, P18 and P60. Right four panels: TEM of transverse cDKO nerve sections …
Figure 4 with 3 supplements
YAP/TAZ are required for proper proliferation of immature SCs.
(A) Quantitation of total SCs (Sox10+ nuclei) per transverse sciatic nerve section in WT and Yap/Taz cDKO mice. n = 3 mice per genotype. ***p<0.001, ****p<0.0001, n.s. = non-significant, 2-way ANOVA …
Figure 4—figure supplement 1
Schwann cell proliferation during postnatal development.
(A) Examples of transverse sections of P0 and P18 WT and cDKO sciatic nerves immunostained to quantify SCs. SC nuclei (green, marked by Sox10). The tibial nerve is delineated by anti-neurofilament …
Figure 4—figure supplement 2
YAP/TAZ are required for integrin α6 expression in Schwann cells.
(A) Transverse sections of P60 WT and Yap/Taz cDKO sciatic nerves immunostained for Integrin α6 (red), neurofilament (green) and DAPI (blue). Integrin α6 localizes to Schwann cells and perineurial …
Figure 4—figure supplement 3
Neuromuscular junctions of Yap/Taz cDKO mice.
(A–A’’’) Two representative junctions from a P55 extensor digitorum longus (EDL) muscle of WT mouse. NF and SV2 immunostaining (green) reveal pre-terminal axons (inset) and axon terminals at …
Figure 5
YAP/TAZ are required for Krox20 upregulation in differentiating Schwann cells.
(A) Identification of Oct6+ SC nuclei at P4 and P60 in WT and Yap/Taz cDKO, as determined by co-staining for Oct6 (red), Sox10 (SC nuclei; green) and DAPI (all cell nuclei; blue). (B) Identification …
Figure 6 with 1 supplement
YAP/TAZ are required for myelin maintenance.
(A) Living eight week-old WT and Yap/Taz iDKO (Sox10-Cre-ERT2) mice 20 days post-first tamoxifen injection. Arrows indicate abnormal splayed gait. (B) Cartoon showing timeline of tamoxifen injection …
Figure 6—figure supplement 1
YAP and TAZ are not expressed in mature oligodendrocytes.
(A) Cartoon denoting a spinal cord white matter area of P60 WT mouse from which quadruple immunostaining images of B–B’’’’ were taken. (B’–B””) Cells strongly immunopositive for YAP/TAZ are present …
Figure 7
Inducible ablation of Yap/Taz in adult Schwann cells leads to rapid loss of Krox20.
(A, B) Immunostaining of longitudinal cryosections of sciatic nerves of WT and iDKO (Sox10-Cre-ERT2) mice 20 days after first tamoxifen injection. (A) Occasional nuclei, marked with DAPI (blue), …
Figure 8 with 1 supplement
YAP/TAZ-TEAD1 regulate Krox20 transcription during developmental myelination and adult myelin maintenance.
(A) Schematic showing potential TEAD binding sites in the Krox20 MSE, in relation to published binding sites for known MSE-binding transcription factors. The regions of the MSE covered by primer …
Figure 8—figure supplement 1
Annotated sequence of the Krox20 Myelin Specific Enhancer.
Sequence shown is the 1 kb distal myelin specific enhancer (MSE) of the mouse Krox20 gene. Sequences covered by primer pairs E1 to E6 used for ChIP qPCR are alternately highlighted in yellow or blue;…
Author response image 1
Videos
Video 1
A movie showing a P60 Yap/Taz cDKO mouse.
https://doi.org/10.7554/eLife.20982.008
Video 2
A movie showing an adult Yap/Taz iDKO and a control littermate mouse.
The iDKO mouse was recorded at 20 days after the first tamoxifen administration.
