J Korean Med Sci.  2009 Aug;24(4):654-659. 10.3346/jkms.2009.24.4.654.

Nidogen Plays a Role in the Regenerative Axon Growth of Adult Sensory Neurons Through Schwann Cells

• Lee HK
• Seo IA
• Suh DJ
• Park HT

• KMID: 1779199
• DOI: http://doi.org/10.3346/jkms.2009.24.4.654

Abstract

We previously reported that nidogen is an extracellular matrix protein regulating Schwann cell proliferation and migration. Since Schwann cells play a critical role in peripheral nerve regeneration, nidogen may play a role in it via regulation of Schwann cells. Here, we demonstrate direct evidence that nidogen induces elongation of regenerative axon growth of adult sensory neurons, and that the effect is Schwann cell dependent. Continuous infusion of recombinant ectodomain of tumor endothelial marker 7, which specifically blocks nidogen function in Schwann cells, suppressed regenerative neurite growth in a sciatic nerve axotomy model. Taken together, it is likely that nidogen is required for proper regeneration of peripheral nerves after injury.

Keyword

Nidogen; Neurites; Regeneration; Dorsal Root Ganglion; Tumor Endothelial Marker 7; Schwann Cells

MeSH Terms

Animals
Axotomy
Cell Movement
Cell Proliferation
Male
Membrane Glycoproteins/*physiology
Membrane Proteins/pharmacology
*Nerve Regeneration
Nerve Tissue Proteins/pharmacology
Neurites/drug effects/*physiology/ultrastructure
Rats
Rats, Sprague-Dawley
Recombinant Proteins/pharmacology
Schwann Cells/cytology/*physiology
Sensory Receptor Cells/*physiology

Figure

• 

  Fig. 1 The effects of Fc and nidogen-Fc on the neurite outgrowth of adult DRG explants. (A) DRG explants from adult rats were cultured in collagen matrix, and the culture medium was supplemented with Fc (20 µg/mL) or nidogen-Fc (20 µg/mL) for 40 hr. Explants are immunostained with anti-Tuj1 to label axons. Scale bar; 100 µm. (B) Quantification of mean length of neurites. Values shown are mean lengths±standard deviation.

• 

  Fig. 2 Schwann cell-mediated neurite outgrowth by nidogen. (A) Dissociated neurons were purified from adult rat DRGs and cultured on Fc or nidogen-Fc-coated dishes for 36 hr in the presence of cytosine arabinoside, then immunostained with an anti-Tuj1 antibody. Representative images of the Fc-control and nidogen-Fc show no significant differences in neurite growth pattern. Scale bar; 10 µm. (B) Quantification of the number of neurite bearing cells in dissociated DRG cultures. (C) DRG explants were cultured on a Millicell membrane for 3 days, and then double immunostained with an antibody to Tuj1 (red) and an antibody to S100 (green). Note the significant colocalization of S100-positive Schwann cells and neurites in the nidogen-Fc treated group. Scale bar; 100 µm.

• 

  Fig. 3 In vivo effect of nidogen inhibition on peripheral nerve regeneration. After sciatic nerve axotomy, AP or eTEM7-AP was infused using an Alzet mini-pump for 2 weeks and then the regenerative axons were stained with an anti-Tuj1 antibody. (A) The axonal morphology is high dystrophic in the eTEM7-treated group compared to the Fc-treated group. Scale bar; 100 µm. (B). The diameter of neurites in eTEM7-AP-treated group was thinner than that of AP controls. *P<0.05. (C). The mean lengths of regenerating neurites from AP and eTEMP7-AP-treated group showed no difference.

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