In vivo trafficking and targeting of N-cadherin to nascent presynaptic terminals

J Neurosci. 2004 Oct 13;24(41):9027-34. doi: 10.1523/JNEUROSCI.5399-04.2004.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Cadherins / genetics*
  • Cadherins / metabolism*
  • Cells, Cultured
  • Embryo, Nonmammalian / cytology
  • Embryo, Nonmammalian / embryology
  • Embryo, Nonmammalian / metabolism
  • Green Fluorescent Proteins / genetics
  • Membrane Proteins / metabolism
  • Microscopy / methods
  • Mutagenesis, Site-Directed
  • Neurons / cytology
  • Neurons / metabolism*
  • Presynaptic Terminals / metabolism*
  • Protein Transport / physiology
  • R-SNARE Proteins
  • Rats
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Sequence Deletion
  • Spinal Cord / cytology
  • Spinal Cord / embryology
  • Spinal Cord / metabolism*
  • Zebrafish

Substances

  • Cadherins
  • Membrane Proteins
  • R-SNARE Proteins
  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins