Transport of cytoskeletal elements from parent axons into regenerating daughter axons.

The kinetics of slow axonal transport in newly regenerating axonal sprouts were compared with those in nonelongating axons. The slowly transported cytoskeletal proteins of ventral motor axons were prelabeled by microinjection of 35S-methionine into the spinal cord. Pulse-labeled slow transport "waves" were observed as they progressed from the surviving "parent" axon stumps (located proximal to a crush lesion) into regenerating "daughter" axon sprouts (located distal to the lesion). Prelabeled cytoskeletal elements of the parent axons were transported into daughter axons, to become distributed into 2 transport waves, "a" and "b." The rate and composition of these waves corresponded to the slow transport subcomponents, SCa and SCb. The shapes of the "a" and "b" waves suggested that the cytoskeletal elements had been reorganized at the junction between the parent and daughter axons. This hypothesis was supported by quantitative analyses of the transport distribution for individual radiolabeled cytoskeletal proteins (actin, spectrin, a 58-67 kDa group that includes microtubule-associated proteins, calmodulin, and tubulin). Specifically, during the first week of outgrowth, the amounts of radiolabeled calmodulin and 58-67 kDa proteins were greater in daughter axons than in nonregenerating control axons. These results support Paul Weiss's "conservative" model of axonal regeneration, which holds that the preexisting transported cytoskeletal elements that continually maintain axonal structure can also provide the cytoskeletal elements required for axonal regeneration. In addition, the results elucidate some of the reorganizational changes in cytoskeletal elements that occur when these are recruited from the parent axon to form daughter axons.