Stability of frog motor nerve terminals in the absence of target muscle fibers

Using repeated in vivo imaging, we addressed the role of target muscle fibers in the maintenance of frog motor nerve terminals at synaptic sites. Target-deprived nerve terminals were generated by selective and permanent removal of muscle fibers without damage to the innervation. Individual nerve terminals, stained with the dye FM1-43, were imaged before and again during the subsequent 1-9 months of target deprivation and the stability of the nerve terminal arbors over time was determined. Repeated observation of motor nerve terminals showed that nerve terminals were well maintained at synaptic sites during the first 1-2 months after target loss; the original number of nerve terminal segments was retained at 85% of the synaptic sites after muscle damage. After long periods of target deprivation, 6-9 months, loss or retraction of nerve terminal segments resulted in a reduction in the arbor of most but not all nerve terminals. This apparent nerve terminal destabilization was not a result of illumination or irradiation because a similar decrease in the extent of nerve terminal arbors was not observed at control irradiated neuromuscular junctions. The persistence of many complete target-deprived nerve terminal arbors at synaptic sites long after target degeneration suggested that the cues that confer stability to frog motor nerve terminals likely reside external to muscle fibers and may be associated with the synaptic basal lamina or the terminal Schwann cell. Since the arbors of many target-deprived nerve terminals were eventually reduced, the nonmuscle stabilization cues may not persist indefinitely at target-deprived synaptic sites.