Journal: The Journal of Neuroscience
Article Title: The Interaction between Tropomyosin-Related Kinase B Receptors and Presynaptic Muscarinic Receptors Modulates Transmitter Release in Adult Rodent Motor Nerve Terminals
Figure Lengend Snippet: Immunolocalization of BDNF, NT-4, trkB, and p75 NTR at the neuromuscular synapses. A–D , Triple labeling of the neurotrophins BDNF ( A ) and NT-4 ( B ) and their receptors trkB ( C ) and p75 NTR ( D ) (green fluorescence) with syntaxin (blue fluorescence) and AChR-α bungarotoxin (red fluorescence). We used plastic-embedded semithin (0.5 to 0.7 μm) cross-sections (STS) as a tool for high-resolution, triple-labeling immunofluorescence analysis. E , Example of triple labeling of syntaxin (green fluorescence), S-100 (blue fluorescence), and nAChR-α bungarotoxin (red fluorescence), which proves that the molecular markers of the three synaptic cells are separated well. F , High-resolution images of synaptic boutons with the terminal axon labeled for one of the proteins in green in the nerve terminal area between the blue Schwann cell and the red nAChR cluster. Scale bars: A–E , 10 μm; F , 1 μm.
Article Snippet: Stock solutions used were as follows: anti-BDNF (G2610; Promega), 50 μg/ml; anti-NT-4 (AB1781; Millipore Bioscience Research Reagents), 50 μg/ml; anti-trkB (clone 47/trkB; 610102; BD Transduction Laboratories), 250 μg/ml; human BDNF (Alomone Labs), 0.37 μ m ; human NT-4 (Alomone Labs), 40 n m ; K-252a (Calbiochem), 5 m m in DMSO; K-252b (Calbiochem), 5 m m ; Pep5 (Calbiochem), 100 μ m ; recombinant human trkB/Fc Chimera (trkB-Fc; 688-TK; R & D Systems), 100 μg/ml.
Techniques: Labeling, Fluorescence, Immunofluorescence