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Fig. 1 | BMC Biology

Fig. 1

From: Functional regeneration of the murine neuromuscular synapse relies on long-lasting morphological adaptations

Fig. 1

Alterations in NMJ morphology after distal chronic denervation of the LAL muscle. a Experimental design for the degenerative model. Approximately 5 mm of the posterior auricular branch of the facial nerve were resected. b Whole mount LAL muscles from control adult mice and mice 7, 30, 45, 60, and 90 days after nerve damage were dissected and processed for immunofluorescence staining with 2H3 (neurofilaments) plus SV2 (synaptic vesicles) antibodies to reveal motor axons and presynaptic terminals (magenta), with an anti S100B antibody to stain Schwann cells (yellow), and with BTX (cyan) to stain AChRs located at postsynaptic densities. Fragmented postsynaptic domains (arrows), detached tSCs (arrowheads), and tSC projections (empty arrowheads) are indicated. Bar = 100 μm. c Representative images of ectopic AChR aggregates at the indicated times after chronic NMJ denervation, AChR were labeled with BTX. Insets show higher magnification of ectopic AChR aggregates. Bar = 50 μm. d Quantification of the percentage of ectopic AChR aggregates at different times after denervation. e, f Nerve terminal and AChR aggregates area were measured at 7 and 30 days after nerve injury and in control muscles. g The overlap between pre and postsynaptic apparatuses was calculated based on nerve terminal and AChR areas. h tSCs behavior in NMJs of control (left panel), early denervated (middle panel), and 30 days of denervation (right panel) muscles. Lower panels show only the SC channel while the postsynaptic domain is delimited (dashed red line). Representative images show tSCs covering the NMJ (arrows) and tSC/SC located outside the NMJ (arrowheads). Bar = 50 μm. i, j Quantification of tSC per NMJ and tSC at the periphery (i.e., within a 50 μm perimeter from the postsynaptic region) of each NMJ. The results are represented as the mean ± SEM and each individual value (N: 3–4 mice; 2 female plus 1–2 male mice). *p < 0.05, Mann–Whitney test (d), **p < 0.01, ****p< 0.0001, one-way ANOVA test (eg, i, j)

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