Neurotrophic factor-α1 modulates NGF-induced neurite outgrowth through interaction with Wnt-3a and Wnt-5a in PC12 cells and cortical neurons

Mol Cell Neurosci. 2015 Sep;68:222-33. doi: 10.1016/j.mcn.2015.08.005. Epub 2015 Aug 11.


Wnt-3a and Wnt-5a signaling activities inhibit and promote neurite outgrowth, respectively, to regulate dendritic and axonal genesis during neurodevelopment. NF-α1, a neurotrophic factor, has been shown to modulate dendritic remodeling and negatively regulate the canonical Wnt-3a pathway. Here, we investigated whether NF-α1 could modify nerve growth factor (NGF)-induced neurite outgrowth through interaction with Wnt-3a and Wnt-5a in PC12 cells and mouse primary cortical neurons. We showed that NGF-induced neurite outgrowth was inhibited by Wnt-3a, and this inhibition was prevented by NF-α1. Western blot analysis revealed that NF-α1 reduced the expression of both β-catenin in the canonical Wnt-3a pathway and Rho, a downstream effector of Wnt-3a's non-canonical signaling pathway. Treatment of PC12 cells with a ROCK inhibitor prevented the inhibition of NGF-induced neurite outgrowth by Wnt-3a, suggesting that NF-α1 promotes neurite outgrowth in the presence of Wnt-3a by down-regulating its canonical and non-canonical activities. Interestingly, treatment of PC12 cells with Wnt-5a, which formed a complex with NF-α1, induced neurite outgrowth that was enhanced by treatment with the combination of Wnt-5a, NGF, and NF-α1. These effects of NF-α1 on Wnt 3a's and Wnt 5a's regulation of neurite outgrowth in PC12 cells were also demonstrated in primary cultures of mouse cortical neurons. In addition, we showed in PC12 cells that NF-α1 acts by upregulating adenomatous polyposis coli (APC) accumulation at neurite tips, thereby providing positive and negative Wnt-3a/Wnt-5a mediated cues to modulate neurite outgrowth, a process important during neurodevelopment.

Keywords: Carboxypeptidase E; NGF; Neurite retraction; PC12 cells; Wnt-3a; Wnt-5a.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Analysis of Variance
  • Animals
  • Cells, Cultured
  • Cerebral Cortex / cytology*
  • Culture Media, Conditioned / pharmacology
  • Enzyme Inhibitors / pharmacology
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Humans
  • Mice
  • Nerve Growth Factor / genetics
  • Nerve Growth Factor / metabolism
  • Nerve Growth Factor / pharmacology*
  • Neurites / drug effects*
  • Neurons / cytology*
  • Neurons / drug effects
  • Neurons / metabolism
  • PC12 Cells / drug effects
  • Rats
  • Rho Factor / metabolism
  • Time Factors
  • Wnt Proteins / metabolism*
  • Wnt-5a Protein
  • Wnt3A Protein / metabolism*


  • Culture Media, Conditioned
  • Enzyme Inhibitors
  • Rho Factor
  • Wnt Proteins
  • Wnt-5a Protein
  • Wnt3A Protein
  • Wnt5a protein, mouse
  • Green Fluorescent Proteins
  • Nerve Growth Factor