Effects of dynactin disruption and dynein depletion on axonal microtubules

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F J AhmadPeter W Baas

Abstract

We investigated potential roles of cytoplasmic dynein in organizing axonal microtubules either by depleting dynein heavy chain from cultured neurons or by experimentally disrupting dynactin. The former was accomplished by siRNA while the latter was accomplished by overexpressing P50-dynamitin. Both methods resulted in a persistent reduction in the frequency of transport of short microtubules. To determine if the long microtubules in the axon also undergo dynein-dependent transport, we ascertained the rates of EGFP-EB3 "comets" observed at the tips of microtubules during assembly. The rates of the comets, in theory, should reflect a combination of the assembly rate and any potential transport of the microtubule. Comets were initially slowed during P50-dynamitin overexpression, but this effect did not persist beyond the first day and was never observed in dynein-depleted axons. In fact, the rates of the comets were slightly faster in dynein-depleted axons. We conclude that the transient effect of P50-dynamitin overexpression reflects a reduction in microtubule polymerization rates. Interestingly, after prolonged dynein depletion, the long microtubules were noticeably misaligned in the distal regions of axons and failed to enter t...Continue Reading

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Citations

Mar 21, 2013·Trends in Cell Biology·Christin A AlbusMike Fainzilber
Sep 25, 2012·Human Molecular Genetics·Daphney C JeanMark M Black
Jun 19, 2008·Annual Review of Neuroscience·Kurt J De VosChristopher C J Miller
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Jul 6, 2012·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Juliana Alves-SilvaAndreas Prokop
Oct 5, 2012·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Shen LinPeter W Baas
Feb 29, 2008·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Joanna M SolowskaPeter W Baas
Nov 5, 2010·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Mei LiuPeter W Baas
Sep 12, 2007·The Journal of Cell Biology·Kenneth A Myers, Peter W Baas
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Jul 23, 2015·Neural Regeneration Research·Peter W Baas, Andrew J Matamoros
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Jun 25, 2013·The International Journal of Biochemistry & Cell Biology·Bàrbara FlixEduard Gallardo
May 4, 2016·The Journal of Cell Biology·Anand N RaoPeter W Baas

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