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The Journal of Neuroscience, May 15, 2002, 22(10):4066-4079
N-Cadherin Mediates Axon-Aligned Process Growth and Cell-Cell Interaction in Rat Schwann Cells
Ina B. Wanner and Patrick M. Wood The Miami Project to Cure Paralysis, and Department of Neurological Surgery, University of Miami School of Medicine, Miami, Florida 33136
The molecular mechanisms underlying the contact behavior of Schwann cells (SCs) and SC-axon association are poorly understood. SC-SC and SC-axon interactions were studied using purified adult rat SCs and cocultures of SCs with embryonic dorsal root ganglion neurons. After contact of SCs with axons, SCs start to extend processes in alignment with axons. This unique alignment was quantitated using a new assay. SC-axon alignment and SC-SC band formation were disrupted in medium containing low extracellular calcium, indicating the involvement of calcium-dependent adhesion molecules. N-cadherin expression was strong in developing rat sciatic nerves but weak in adult sciatic nerves. In purified adult-derived rat SCs, N-cadherin expression was increased by mitogens (neuregulins) and decreased by high cell density. High-resolution confocal images show intense N-cadherin signals in SC process tips. Subcellular N-cadherin was accumulated in bands at intercellular junctions between SCs and was clustered at axon-SC contact sites. Blocking antibodies (rabbit and guinea pig IgG directed against the first extracellular domain of N-cadherin) and cyclic pentapeptides (including the HAV motif) were used to perturb N-cadherin function. All blocking agents reduced the number of N-cadherin-positive SC-SC junctions and perturbed axon-aligned growth of SC processes. Averaging over all N-cadherin-perturbation experiments, in controls 67-86% of SCs exhibited axon-aligned process growth, whereas in treated cultures only 41% of the SCs aligned with axons. These results are evidence that in mammals N-cadherin is important for formation of SC-SC junctions and SC process growth in alignment with axons.
Key words: cell adhesion molecule; N-cadherin; neuregulin; axon-glia interaction; subcellular localization; function-blocking agents; Schwann cell-DRG neuron coculture
Copyright © 2002 Society for Neuroscience 0270-6474/02/22104066-14$05.00/0
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