Neuromuscular junctions (NMJs) of Eps15 homology domain-containing (EHD)1−/−mice are innervated by a single motor axon. Panel (A) Isolated tibialis anterior (TA) skeletal muscle fibers from three EHD1−/− and three wild-type (WT) mice were labeled with (BTX) Alexa Fluor 555 conjugate for morphological mapping of endplates using confocal microscopy. Rare EHD1−/−endplates (n = 2) consisted of two discreet structures, with one being smaller and less developed than the other. (B) Isolated TA skeletal muscle fibers from three EHD1−/− and three WT mice were immunolabeled with neuronal class III β-tubulin monoclonal antibody (TUJ1) antibody, a neuronal marker, and visualized using confocal microscopy. WT (n = 13) and EHD1−/− (n = 12) endplates with partitioning were seen. EHD1−/− endplates were singly innervated, even when partitioned. Blue, DAPI; green, α-BTXAlexa Fluor 488 conjugate; red, anti-mouse Alexa Fluor 568 conjugate. Scale bars: 20 μm. A larger reference bar was added above the scale bars.