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The Journal of Neuroscience, October 13, 2004, 24(41):9027-9034; doi:10.1523/JNEUROSCI.5399-04.2004

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Cellular/Molecular
In Vivo Trafficking and Targeting of N-Cadherin to Nascent Presynaptic Terminals

James D. Jontes, Michelle R. Emond, and Stephen J Smith

Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, California 94305

N-cadherin is a prominent component of developing and mature synapses, yet very little is known about its trafficking within neurons. To investigate N-cadherin dynamics in developing axons, we used in vivo two-photon time-lapse microscopy of N-cadherin—green fluorescent protein (Ncad-GFP), which was expressed in Rohon-Beard neurons of the embryonic zebrafish spinal cord. Ncad-GFP was present as either stable accumulations or highly mobile transport packets. The mobile transport packets were of two types: tubulovesicular structures that moved preferentially in the anterograde direction and discrete-punctate structures that exhibited bidirectional movement. Stable puncta of Ncad-GFP accumulated in the wake of the growth cone with a time course. Colocalization of Ncad-GFP puncta with synaptic markers suggests that N-cadherin is a very early component of nascent synapses. Expression of deletion mutants revealed a potential role of the extracellular domain in appropriate N-cadherin trafficking and targeting. These results are the first to characterize the trafficking of a synaptic cell-adhesion molecule in developing axons in vivo. In addition, we have begun to investigate the cell biology of N-cadherin trafficking and targeting in the context of an intact vertebrate embryo.

Key words: adhesion; imaging; spinal; synaptogenesis; Rohon-Beard; in vivo

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Received Dec 6, 2003; revised August 26, 2004; accepted August 31, 2004.

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