Evidence that myosin activity opposes microtubule-based axonal transport of mitochondria.
Neurons transport and position mitochondria using a combination of saltatory, bidirectional movements and stationary docking. Axonal mitochondria move along microtubules (MTs) using kinesin and dynein motors, but actin and myosin also play a poorly defined role in their traffic. To ascertain this role, we have used RNA interference (RNAi) to deplete specific myosin motors in cultured Drosophila neurons and quantified the effects on mitochondrial motility. We produced a fly strain expressing the Caenorhabditis elegans RNA transporter SID-1 in neurons to increase the efficacy of RNAi in primary cultures. These neurons exhibited significantly increased RNAi-mediated knockdown of gene expression compared with neurons not expressing this transporter. Using this system, we observed a significant increase in mitochondrial transport during myosin V depletion. Mitochondrial mean velocity and duty cycle were augmented in both anterograde and retrograde directions, and the fraction of mitochondrial flux contained in long runs almost doubled for anterograde movement. Myosin VI depletion increased the same movement parameters but was selective for retrograde movement, whereas myosin II depletion produced no phenotype. An additional effect of myosin V depletion was an increase in mitochondrial length. These data indicate that myosin V and VI play related but distinct roles in regulating MT-based mitochondrial movement: they oppose, rather than complement, protracted MT-based movements and perhaps facilitate organelle docking.
Pathak
Divya
D
Department of Biological Sciences and Purdue University Integrative Neuroscience Program, West Lafayette, IN 47907, USA.
Sepp
Katharine J
KJ
Hollenbeck
Peter J
PJ
D000818:Animals; D030801:Animals, Genetically Modified; D001370:Axonal Transport / Q000502:physiology*; D017173:Caenorhabditis elegans; D029742:Caenorhabditis elegans Proteins / Q000235:genetics / Q000378:metabolism; D002478:Cells, Cultured; D004330:Drosophila; D055785:Gene Knockdown Techniques; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D008870:Microtubules / Q000502:physiology*; D008928:Mitochondria / Q000502:physiology*; D009068:Movement; D018995:Myosin Heavy Chains / Q000235:genetics / Q000378:metabolism*; D024462:Myosin Type II / Q000235:genetics / Q000378:metabolism*; D024701:Myosin Type V / Q000235:genetics / Q000378:metabolism*; D009474:Neurons / Q000502:physiology; D010641:Phenotype; D034622:RNA Interference
D029742:Caenorhabditis elegans Proteins; D008565:Membrane Proteins; C452203:SID-1 protein, C elegans; C089785:myosin VI; D024462:Myosin Type II; D024701:Myosin Type V; D018995:Myosin Heavy Chains