These authors contributed equally to this work.
In vivo Imaging of Retrogradely Transported Synaptic Vesicle Proteins in Caenorhabditis elegans Neurons
Article first published online: 29 OCT 2010
© 2010 John Wiley & Sons A/S
Volume 12, Issue 1, pages 89–101, January 2011
How to Cite
Murthy, K., Bhat, J. M. and Koushika, S. P. (2011), In vivo Imaging of Retrogradely Transported Synaptic Vesicle Proteins in Caenorhabditis elegans Neurons. Traffic, 12: 89–101. doi: 10.1111/j.1600-0854.2010.01127.x
- Issue published online: 9 DEC 2010
- Article first published online: 29 OCT 2010
- Accepted manuscript online: 4 OCT 2010 12:21PM EST
- Received 3 March 2010, revised and accepted for publication 1 October 2010, uncorrected manuscript published online 4 October 2010, published online 29 October 2010
- dyenin–dynactin mutants;
- retrograde axonal transport;
- synaptic vesicle proteins;
Axonal transport is an essential process that carries cargoes in the anterograde direction to the synapse and in the retrograde direction back to the cell body. We have developed a novel in vivo method to exclusively mark and dynamically track retrogradely moving compartments carrying specific endogenous synaptic vesicle proteins in the Caenorhabditis elegans model. Our method is based on the uptake of a fluorescently labeled anti-green fluorescent protein (GFP) antibody delivered in an animal expressing the synaptic vesicle protein synaptobrevin-1::GFP in neurons. We show that this method largely labels retrogradely moving compartments. Very little labeling is observed upon blocking vesicle exocytosis or if the synapse is physically separated from the cell body. The extent of labeling is also dependent on the dyenin–dynactin complex. These data support the interpretation that the labeling of synaptobrevin-1::GFP largely occurs after vesicle fusion and the major labeling likely takes place at the synapse. Further, we observe that the retrograde compartment carrying synaptobrevin contains synaptotagmin but lacks the endosomal marker RAB-5. This labeling method is very general and can be readily adapted to any transmembrane protein on synaptic vesicles with a GFP tag inside the vesicle and can also be extended to other model systems.