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Fig. 5 | Molecular Neurodegeneration

Fig. 5

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

Fig. 5

Morphology and time course of neuromuscular synapses after crush injury of the sciatic nerve. a Overview of alpha-bungarotoxin-Alexa Fluor 594 (αBGX) stained neuromuscular junctions (light dots) in muscle sections of the ipsilateral gastrocnemius muscles after crush injury of the sciatic nerve of SLICK-Grn mice (without tamoxifen) and SLICK-Grn-OE mice (with tamoxifen). b Examples of the time course of neuromuscular junctions. The presynaptic site was identified by immunostaining of the synaptic vesicle protein, SV2; the postsynaptic site by labeling of nicotinic acetylcholine receptors with αBGX. Scale bars 200 μm. c Examples of neuromuscular junctions 21 and 42days after nerve injury. NMJs were identified via EYFP and αBGX to detect the nicotinic acetylcholine receptor. The EYFP signal arises from the regrowing axon and shows the presynaptic site. The gross morphology of the neuromuscular junctions was similar in control and tamoxifen treated progranulin overexpressing mice, but the number of intact NMJs was higher in the latter. Scale bars 50 μm. d Counts of αBGX-positive neuromuscular junctions in gastrocnemius muscles after crush injury of the sciatic nerve per muscle area (means ± SD). The contralateral sides served as controls (all time points summarized). The asterisks (*) and crosses (#) mark time dependent significant changes in the counts of ipsilateral NMJs per square μm of the muscle versus contralateral. The circle shows a significant difference between groups. Thirty to forty sections were analyzed per group, per time point and represent results of 5–6 mice per group and time point. P < 0.05 for all tests. e Erk phosphorylation (pErk immunofluorescence) in primary DRG neurons stimulated with 50 nM nerve growth factor, NGF or 10 ng/ml recombinant human progranulin (rPGRN) in the absence of NGF. The number of pErk positive and NeuN positive neurons were counted per frame, 2–3 frames per culture (Axiovision autmess software). Data are means ± SD percentages of pErk + neurons. Asterisks show significant increases of pErk positive neurons versus unstimulated neurons (one-way ANOVA, P < 0.05, posthoc Dunnett versus unstimulated). rPGRN and NGF had similar effects

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