Filopodia, lamellipodia and retractions at mouse neuromuscular junctions

In order to determine if mature motor nerve terminals retain structures associated with development such as filopodia and lamellipodia, we studied whole mounts of mature mouse neuromuscular junctions stained with both fluorescent-labelled tetanus toxin C-fragment and alpha-bungarotoxin, and employed electron microscopy in parallel. The rapid fluorescent stain may be of general usefulness. Both filopodia and lamellipodia were found, extending beyond the border of the established postsynaptic receptors. Filopodia often appeared in clusters, were devoid of a synaptic vesicle antigen, and many withdrew in response to cytochalasin D. Control experiments demonstrated that filopodia were not induced by the toxin treatment. The mean number of filopodia per endplate varied from about one in phasic muscle to three in tonic muscle, and was twice as great in immature mouse muscle. Postsynaptic receptor-rich regions without overlying terminals were less numerous than filopodial and lamellipodial projections without underlying receptors. Electron microscopy showed that lamellipodia contained actin-like filaments and immunoreactivity to actin, but no neurofilaments, microtubules, mitochondria or vesicles. Therefore, these structures would not be visualized by in vivo mitochondrial stains. The lamellipodia protruded into the gap between muscle and a closely overlying Schwann cell process. Lamellipodia occupied about 5% of the linear extent of the terminal arbor in whole mounts, but appeared in 16% of random electron micrographic fields. Thus, the lamellipodia and filopodia typical of developing terminals are present in adulthood and represent a distinctive specialization of the nerve terminal, which may interact with the adjacent Schwann and muscle cell. The frequency of filopodia is a function of age and of muscle or motoneuron type. We suggest that some of the factors known to regulate growth of filopodia and lamellipodia in vitro or in development may continue to act at adult presynaptic nerve terminals.