Retrograde but not anterograde bead movement in intact axons requires dynein.
Dynein and kinesin have been implicated as the molecular motors that are responsible for the fast transport of axonal membranous organelles and vesicles. Experiments performed in vitro with partially reconstituted preparations have led to the hypothesis that kinesin moves organelles in the anterograde direction and dynein moves them in the retrograde direction. However, the molecular basis of transport directionality remains unclear. In the experiments described here, carboxylated fluorescent beads were injected into living Mauthner axons of lamprey and the beads were observed to move in both the anterograde and retrograde directions. The bead movement in both directions required intact microtubules, occurred at velocities approaching organelle fast transport in vivo, and was inhibited by vanadate at concentrations that inhibit organelle fast transport. When living axons were injected with micromolar concentrations of vanadate and irradiated at 365 nm prior to bead injections, a treatment that results in the V1 photolysis of dynein, the retrograde movement of the beads was specifically abolished. Neither the ultraviolet irradiation alone nor the vanadate alone produced the retrograde-specific inhibition. These results support the hypothesis that dynein is required for retrograde, but not anterograde, transport in vivo.
Wang
C
C
Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907-1392, USA.
Asai
D J
DJ
Robinson
K R
KR
D000818:Animals; D001370:Axonal Transport / Q000187:drug effects / Q000502:physiology* / Q000528:radiation effects; D003078:Colchicine / Q000494:pharmacology; D004398:Dyneins / Q000502:physiology*; D005453:Fluorescence; D007091:Image Processing, Computer-Assisted; D007798:Lampreys / Q000502:physiology*; D007814:Larva / Q000502:physiology; D008845:Microinjections; D008856:Microscopy, Fluorescence; D018715:Microscopy, Video; D008863:Microspheres; D008870:Microtubules / Q000502:physiology; D013116:Spinal Cord / Q000166:cytology / Q000187:drug effects / Q000528:radiation effects; D014466:Ultraviolet Rays; D014638:Vanadates / Q000494:pharmacology
D014638:Vanadates; D004398:Dyneins; D003078:Colchicine