Journal: The Journal of Neuroscience

Article Title: The Agrin/MuSK Signaling Pathway Is Spatially Segregated from the Neuregulin/ErbB Receptor Signaling Pathway at the Neuromuscular Junction

doi: 10.1523/jneurosci.20-23-08762.2000

Figure Lengend Snippet: Figure 1. Immunofluorescence visualiza- tion of proteins enriched at the NMJ. Pro- teins of the agrin/MuSK and neuregulin/ erbB signaling pathways as well as marker proteins for subdomains of the NMJ were visualized in cross sections of adult rat gastrocnemius muscle using a 203 objec- tive. Bodipy-aBtx was used to identify in- dividual NMJs. The axon terminal marker SV2, Schwann cell marker S-100, utrophin (a protein enriched in the postsynaptic pri- mary gutter), and dystrophin (a protein enriched in the secondary folds) all show clear staining at the NMJ. Dystrophin ad- ditionally shows the expected enrichment along the sarcolemma. Immunofluores- cence from antibodies against erbB2, erbB3, erbB4, neuregulin, agrin, and MuSK all show enrichment at the NMJ with little extra-junctional staining of the muscle. Scale bar, 200 mm.

Article Snippet: The mouse monoclonal anti-erbB3 (05-390) and rabbit polyclonal anti-erbB4 (06–572) antibodies were from Upstate Biotechnology (Lake Placid, NY); each was used at a 1:100 dilution.

Techniques: Protein-Protein interactions, Marker, Staining

Journal: The Journal of Neuroscience

Article Title: The Agrin/MuSK Signaling Pathway Is Spatially Segregated from the Neuregulin/ErbB Receptor Signaling Pathway at the Neuromuscular Junction

doi: 10.1523/jneurosci.20-23-08762.2000

Figure Lengend Snippet: Figure 3. Confocal immunofluorescence analysis of erbB receptor distri- butions at adult rat gastrocnemius NMJ. En face (A1, D1, G1) and side view (B1, E1, H1) images of NMJs stained with Bodipy-aBtx. The NMJs were double-labeled with antibodies against erbB2 (A2, B2), erbB3 (D2, E2), and erbB4 (G2, H2) and visualized using a Cy3-conjugated secondary antibody. Color overlays are shown with a-Btx in green and the respective antibody in red (A3–H3). Schematics of these distributions are shown in C, F, and I. ErbB2 is enriched in the secondary folds. ErbB3 is enriched in the Schwann cell. A gap between the primary gutter and the erbB3 immunoreactivity can be seen in E3 (arrow). ErbB4 is enriched in the secondary folds. Scale bar, 10 mm.

Article Snippet: The mouse monoclonal anti-erbB3 (05-390) and rabbit polyclonal anti-erbB4 (06–572) antibodies were from Upstate Biotechnology (Lake Placid, NY); each was used at a 1:100 dilution.

Techniques: Staining, Labeling