Impairment of anterograde and retrograde neurofilament transport after anti-kinesin and anti-dynein antibody microinjection in chicken dorsal root ganglia.
The purpose of the present study was to investigate the participation of the motor proteins kinesin and dynein in axonal transport of neurofilaments (NF) in cultured dorsal root ganglia neurons. Therefore, we performed live-recording studies of the green fluorescent protein-tagged neurofilament M (GFP-NF-M) to assay transport processes in neurons. Co-localization studies revealed that GFP-NF-M was capable to build a functional NF network with other NF subunits, including phosphorylated heavy neurofilaments (NF-H-PH). Time-lapse recordings using confocal laser scanning microscopy exhibited fast transport of NF dots in anterograde and retrograde direction through a photobleached gap. Following microinjection of anti-kinesin antibodies or colchicine treatment an impairment of anterograde as well as retrograde NF transport was observed during live-recording experiments. In contrast, microinjection of anti-dynein antibodies only impaired retrograde transport of NF whereas the anterograde movement of GFP-NF-M was unaffected. Treatment of the cells with unspecific antibodies had no effect.
Theiss
Carsten
C
Department of Cytology, Institute of Anatomy, Faculty of Medicine, Ruhr-University Bochum, Universitätsstr 150, D-44780 Bochum, Germany. carsten.theiss@ruhr-uni-bochum.de
Napirei
Markus
M
Meller
Karl
K
D000818:Animals; D015153:Blotting, Western; D001921:Brain / Q000196:embryology*; D002642:Chick Embryo; D003001:Cloning, Molecular; D003078:Colchicine / Q000494:pharmacology; D018076:DNA, Complementary / Q000378:metabolism; D004398:Dyneins / Q000737:chemistry*; D005727:Ganglia, Spinal / Q000378:metabolism*; D006074:Gout Suppressants / Q000494:pharmacology; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D007382:Intermediate Filaments / Q000378:metabolism; D016547:Kinesins / Q000737:chemistry* / Q000378:metabolism; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D016900:Neurofilament Proteins / Q000378:metabolism; D009474:Neurons / Q000378:metabolism; D010766:Phosphorylation; D011312:Pressure; D011485:Protein Binding; D051381:Rats; D013997:Time Factors
D018076:DNA, Complementary; D006074:Gout Suppressants; D016900:Neurofilament Proteins; D049452:Green Fluorescent Proteins; D004398:Dyneins; D016547:Kinesins; D003078:Colchicine