Journal: Frontiers in Molecular Neuroscience
Article Title: Arhgef5 Binds α-Dystrobrevin 1 and Regulates Neuromuscular Junction Integrity
Figure Lengend Snippet: Generation of muscle-specific Arhgef5 KO mice. (A) Strategy for the generation of conditional Arhgef5 knockouts. Mice with loxP sites flanking the third exon of Arhgef5 were crossed with Cre-recombinase expressing mice to produce knockout. (B) Genotyping shows successful recombination in the muscles but not tail tipss of AG5 fl/fl ; Acta-Cre mice. (C) Quantification of Arhgef5 mRNA expression in AG5 fl/fl and AG5 fl/fl ; Acta-Cre mice at 50 and 500 days of age. Results are mean ± SEM of three mice from each genotype. (D) Proper alignment of pre- and post-synaptic elements in AG5 fl/fl ; Acta-Cre muscles. Single fibers of tibialis anterior muscles isolated from AG5 fl/fl and AG5 fl/fl ; Acta-Cre mice were stained with BTX (to visualize AChR), fasciculin II (to visualize the synaptic cleft marker acetylcholinesterase), and anti-neurofilament + anti-synaptophysin antibodies (to visualize the presynaptic nerve terminal). Scale bar is 20 μm. ∗ p
Article Snippet: For visualization of AChR we used fluorescently labeled α-bungarotoxin (B35451 or B13422; Thermo), and for visualizing the synaptic cleft, we used fasciculin II, which binds to acetylcholinesterase (F-225; Alomone Labs, labeled with FluoReporter FITC kit from Thermo).
Techniques: Mouse Assay, Expressing, Knock-Out, Isolation, Staining, Marker