Rab18−/− mice have large accumulations of neurofilament at the neuromuscular junction. Rab18−/− mice have large accumulations of neurofilament at the neuromuscular.

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Rab18−/− mice have large accumulations of neurofilament at the neuromuscular junction. Rab18−/− mice have large accumulations of neurofilament at the neuromuscular junction. (A–F) Confocal micrographs showing the neuromuscular junction in (A,D) flexor digitorum brevis (FDB), (B,E) lumbrical and (C,F) transverse abdominus muscle preparations from control (A–C) and Rab18−/− mice (D–F). TRITC-conjugated α-bungarotoxin staining is shown in red, staining of 165-kDa neurofilament and synaptic vesicle protein SV2 are both shown in green. (G–I) Bar charts (mean±s.e.m.) showing that, in all muscles examined – (G) FDB, (H) lumbrical and (I) TVA – the majority of endplates from both early- and mid-late-symptomatic Rab18−/− mice were fully occupied (where SV2 staining overlayed the endplate). Statistical significance was assessed using a Mann–Whitney U test. Early symptomatic: FDB P-value=0.5553, muscles from control mice (n=8) and muscles from Rab18−/− mice (n=7); lumbrical P-value=0.8120, muscles from control mice (n=10) and muscles from Rab18−/− mice (n=8); TVA muscle (n=1). Mid-late symptomatic: FDB P-value=0.3523, muscles from control mice (n=9) and muscles Rab18−/− mice (n=8); lumbrical P-value=1.0, muscles (n=10); TVA P-value=1.0, muscles (n=3). ns, not significant. (J–L) Bar charts (mean±s.e.m.) showing the percentage of endplates that exhibited large accumulations (stained green) in Rab18−/− mice at early- and mid-late-symptomatic timepoints. Statistical significance was assessed using a Mann–Whitney U test. Early symptomatic: FDB P-value=0.0026, muscles from control mice (n=8) and muscles from Rab18−/− mice (n=7); lumbrical P-value=0.0096, muscles from control mice (n=10) and muscles from Rab18−/− mice (n=8); TVA muscle (n=1). Mid-late symptomatic: FDB P-value=0.0005, muscles from control mice (n=9) and muscles from Rab18−/− mice (n=8); lumbrical P-value=0.0002, muscles (n=10); TVA P-value=0.0765, muscles (n=3). ns, not significant, **P<0.005, ***P<0.001. (M–R) Confocal micrographs showing neuromuscular junctions from Rab18−/− lumbrical muscles that had been immunostained with TRITC-conjugated α-bungarotoxin (red) and one of either neurofilament (NEFM) (M–O) or SV2 (P–R). Note the large neurofilament accumulations in Rab18−/− endplates (O) but normal levels of synaptic vesicle marker SV2 (R). Scale bars: 10 μm. Sarah M. Carpanini et al. Dis. Model. Mech. 2014;7:711-722 © 2014. Published by The Company of Biologists Ltd