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Fig. 2 | Molecular Neurodegeneration

Fig. 2

From: Progranulin promotes peripheral nerve regeneration and reinnervation: role of notch signaling

Fig. 2

Immunofluorescent analysis of axonal regrowth and motor neuron survival after injury of the sciatic nerve. a Cyclic GMP dependent protein kinase (PKG1) immunofluorescence 1, 3 and 7 days after crush injury of the sciatic nerve in SLICK-Grn mice (without tamoxifen) and SLICK-Grn-OE mice (with tamoxifen). PKG1 is a marker of axonal transport and regrowth. The images are rainbow pseudo-color images of fluorescent intensities. The left panels are in situ hybridizations showing progranulin mRNA expression of the respective DRGs and ventral horns of the spinal cord. The figures show representative results of n = 3 per time point. Scale bars 100 μm ventral horn, 50 μm DRGs and 200 μm nerves. b 2D intensity profile plots showing the quantitative analysis of PKG1-immunofluorescence in the sciatic nerves (exemplary result of n = 3 per time point). The grey area shows the remaining gap at 7days. PKG1 initially accumulates in front of the lesion and then approaches towards the distal end with the outgrowing fibers, when the axonal transport is resumed. c PKG1 immunofluorescent intensity (rainbow pseudo-color) of the lesion site of the sciatic nerve in adult progranulin deficient and control mice. There was no regrowth and a long gap in Grn−/− mice. Scale bar 100 μm. Images are examples of 3–4 mice per group. d ATF3 immunofluorescence of injured motor neurons in the ventral horn of the spinal cord of Grn−/− and Grn+/+ mice after sciatic nerve injury in pups showing each 2 representative images. ATF3 is a marker of axonal injury, neurons are stressed but alive. Grn−/− either had a high number of ATF3 positive neurons or had lost these neurons. The bimodal distribution is reflected in the frequency distribution plots in e. e Quantitative analysis of ATF3 positive neurons showing the frequency distribution of ATF3 positive neurons per section. 142 sections of 6 mice were analyzed per group. The number of ATF3+ neurons per section followed a normal distribution in Grn+/+ mice, but was left-skewed in Grn−/− mice showing that ATF3+ neurons had died. In Grn−/− mice, there was a second peak representing sections with high numbers of ATF3+ neurons. The cumulative frequency distributions were statistically compared using the two sample Kolmogorov-Smirnov test. The null hypothesis that both distributions were equal was rejected

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