Dynamics of Kv1 channel transport in axons.
Concerted actions of various ion channels that are precisely targeted along axons are crucial for action potential initiation and propagation, and neurotransmitter release. However, the dynamics of channel protein transport in axons remain unknown. Here, using time-lapse imaging, we found fluorescently tagged Kv1.2 voltage-gated K(+) channels (YFP-Kv1.2) moved bi-directionally in discrete puncta along hippocampal axons. Expressing Kvbeta2, a Kv1 accessory subunit, markedly increased the velocity, the travel distance, and the percentage of moving time of these puncta in both anterograde and retrograde directions. Suppressing the Kvbeta2-associated protein, plus-end binding protein EB1 or kinesin II/KIF3A, by siRNA, significantly decreased the velocity of YFP-Kv1.2 moving puncta in both directions. Kvbeta2 mutants with disrupted either Kv1.2-Kvbeta2 binding or Kvbeta2-EB1 binding failed to increase the velocity of YFP-Kv1.2 puncta, confirming a central role of Kvbeta2. Furthermore, fluorescently tagged Kv1.2 and Kvbeta2 co-moved along axons. Surprisingly, when co-moving with Kv1.2 and Kvbeta2, EB1 appeared to travel markedly faster than its plus-end tracking. Finally, using fission yeast S. pombe expressing YFP-fusion proteins as reference standards to calibrate our microscope, we estimated the numbers of YFP-Kv1.2 tetramers in axonal puncta. Taken together, our results suggest that proper amounts of Kv1 channels and their associated proteins are required for efficient transport of Kv1 channel proteins along axons.
Gu
Yuanzheng
Y
Department of Neuroscience and Center for Molecular Neurobiology, The Ohio State University, Columbus, Ohio, USA.
Gu
Chen
C
D000818:Animals; D001369:Axons / Q000378:metabolism*; D016547:Kinesins / Q000378:metabolism; D051663:Kv1.2 Potassium Channel / Q000737:chemistry / Q000378:metabolism*; D008164:Luminescent Proteins / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D057054:Molecular Imaging; D009068:Movement; D016296:Mutagenesis; D009154:Mutation; D024642:Potassium Channels, Voltage-Gated / Q000235:genetics / Q000378:metabolism; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D051381:Rats
C426239:Kcnab2 protein, rat; C490201:Kif3a protein, rat; D051663:Kv1.2 Potassium Channel; D008164:Luminescent Proteins; D008869:Microtubule-Associated Proteins; D024642:Potassium Channels, Voltage-Gated; D016547:Kinesins