Cytoplasmic dynein conversion at a crush injury in rat peripheral axons
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Cytoplasmic dynein is a motor for retrograde axonal transport for movement of membranous organelles toward the neuronal cell body, However, cytoplasmic dynein is synthesized in the cell body and conveyed along the axon to nerve terminals. To characterize the axonal transport of cytoplasmic dynein in relation to synaptic vesicles and other membrane compartments, immunocytochemical and cytofluorimetric scanning analyses of crush-operated rat sciatic nerves were performed. Distal to the crush, the kinetics of dynein accumulation were consistent with its role in the retrograde transport of membranous organelles. During the initial 3 hr after crush, only small amounts of dynein-immunoreactive material accumulated proximal to the crush. This is consistent with metabolic labeling studies showing that most of the dynein moving in the anterograde direction is in the slow component of axonal transport. Thereafter, the rate of proximal accumulation of dynein increased, and by 8 hr postcrush a large amount of dynein immunoreactivity was observed. This accelerated accumulation may be due to recruitment of dynein from slow component b onto organelles proximal to the crush. Double labeling demonstrated that dynein immunoreactivity colocalized with synaptophysin, a transmembrane protein found in small, clear synaptic vesicles. In contrast, dynein immunoreactivity did not colocalize well with calcitonin gene-related peptide (CGRP), a peptide matrix marker for some large dense-cored vesicles, Finally, dynein immunoreactivity colocalized with the anterograde transport motor kinesin both proximal and distal to a crush, suggesting that kinesin may carry some dynein-containing membrane compartments during fast anterograde axonal transport, (C) 2000 Wiley-Liss, Inc.