Tools for studying the role of N-cadherin mediated extracellular interaction in neuronal development and function

Abstract

N-cadherin is a homophilic cell adhesion molecule that plays important roles in many aspects of neuronal development. In order to better understand the function of N-cadherin mediated cell-cell contact in activity-dependent dendrite development, we generated a number of new tools. EC1, consisting of the first extracellular domain of N-cadherin, can specifically inhibit N-cadherin, but not E-cadherin, mediated cell-cell contact, both when over-expressed in neurons and added as a purified protein. Ncad-HA is an extracellularly epitope-tagged version of N-cadherin that, when over-expressed under the activity-independent pCS2min promoter, can be used to assay surface N-cadherin level following various manipulations. These tools are likely to be very useful for studying the function of N-cadherin in multiple aspects of neural circuit development.

Figure 1

Schematic of N-cadherin and its derived tools. Top line is a domain structure of mature full-length N-cadherin with the critical second tryptophan (W) residue and location of histidine-alanine valine (HAV) peptide illustrated; EC, extracellular domain; TM, transmembrane domain. Middle is EC1-myc, containing the first EC domain of N-cadherin fused to a C-terminal myc tag (green). Bottom is the N-cadherin-HA (N-cad-HA) construct, with an HA tag (red) inserted between EC2 and EC3 domains.