1. Expression and localization of Wnt components during sciatic nerve development.
Expression and localization of Wnt components during sciatic nerve development. A, Total RNA from sciatic nerves of mice at different developmental stages (P0, P6, and P20) was prepared. qRT-PCR experiments were performed with primers specifically recognizing Wnt ligands (Wnt1, Wnt3, Wnt3a, Wnt7a), Wnt components (LRP6, DSH1, DSH2, DSH3, ß catenin, TCF1, LEF1, TCF3, TCF4), or sequences of the peripheral myelin protein genes MPZ or PMP22. We used 26S proteasomal RNA (26S) to normalize the qRT-PCR data as the expression of 26S RNA was constant at P0, P6, and P21. We established a ratio for each RNA (Wnt1, Wnt3, Wnt3a, Wnt7a, MPZ, PMP22) relative to 26S rRNA at P0 (n = 3). B, The spatial expression patterns of various members of the Wnt signaling pathway in sciatic nerve sections of mice aged 7 d. Myelin sheaths were stained with anti-MBP antibody (in red), and Wnt components were localized with antibodies against Wnt1, LRP6, GSK3β, and β-catenin (in green). The slides were analyzed with a confocal microscope. This experiment was repeated twice, and a typical experiment is presented here. The images shown represent 1 μm of confocal Z sectioning. Scale bars, 2 μm.
Marcel Tawk et al. J. Neurosci. 2011;31:
©2011 by Society for Neuroscience