Journal: eLife

Article Title: Site-directed MT1-MMP trafficking and surface insertion regulate AChR clustering and remodeling at developing NMJs

doi: 10.7554/eLife.54379

Figure Lengend Snippet: ( A ) Representative confocal images showing the localization of endogenous MT1-MMP at perforations of synaptic AChR clusters (arrows, top panels) in longitudinal cryosections of adult rat soleus muscles. The specificity of MT1-MMP antibody was verified by pre-incubating anti-MT1-MMP primary antibody with recombinant human MT1-MMP protein (rhMT1-MMP, bottom panels). Neurofilament (NF) was used as a neuronal marker. ( B ) Representative confocal images showing the postsynaptic localization of MT1-MMP at NMJs as demonstrated by the surgical denervation experiment. Staining of AChR, MT1-MMP, and presynaptic marker (NF and synaptophysin (Syn)) was performed using cross sections from adult rat soleus muscles either without (top panels) or with sciatic nerve cut after 4 days (bottom panels). ( C ) Representative confocal images showing aneural versus synaptic AChR clusters in whole-mount diaphragms from wild-type (WT) control and MT1-MMP -/- mice at E13.5 (left panels) and E18.5 (right panels). Whole-mount tissues were stained for AChR and NF. The superimposed 3D reconstruction images were generated by z-stack images using Imaris. Synaptic AChR clusters (yellow) were identified when signals of AChR and NF overlapped with each other, whereas other AChR signals (red) represent aneural AChR clusters. ( D–G ) Quantification on the density of aneural ( D ) versus synaptic ( E ) AChR clusters, the width of end-plate bands ( F ), and the length of axonal branches ( G ) in diaphragm muscles between wild-type and MT1-MMP -/- mouse embryos at E13.5 and E18.5. n = 4 (E13.5, WT) and 5 (E13.5, MT1-MMP -/- ), 5 (E18.5, WT), and 3 (E18.5, MT1-MMP -/- ) embryos from three independent experiments. Scale bars represent 10 μm ( A ) or 100 μm ( C ). Data are represented as mean ± SEM. Two-way ANOVA with Sidak’s multiple comparison test ( D and E ), Student’s t-test ( F and G ), *, **, *** represent p≤0.05, 0.01, and 0.001 respectively. n.s.: non-significant.

Article Snippet: Peptide, recombinant protein , Recombinant human MT1-MMP , R and D Systems , Cat#: 918-MP-010 , .

Techniques: Muscles, Recombinant, Marker, Staining, Control, Generated, Comparison

Journal: eLife

Article Title: Site-directed MT1-MMP trafficking and surface insertion regulate AChR clustering and remodeling at developing NMJs

doi: 10.7554/eLife.54379

Figure Lengend Snippet:

Article Snippet: Peptide, recombinant protein , Recombinant human MT1-MMP , R and D Systems , Cat#: 918-MP-010 , .

Techniques: Recombinant, Sequencing, Control, Cell Attachment Assay, Software, Microscopy, Imaging, Electron Microscopy