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Expression profiling upon Nex1/MATH‐2‐mediated neuritogenesis in PC12 cells and its implication in regeneration

The expression of Nex1 peaks during brain development when neurite outgrowth and synaptogenesis are highly active. We previously showed that Nex1 is a critical effector of the nerve growth factor (NGF) pathway and its overexpression results in spontaneous neuritogenesis. Furthermore, the PC12‐Nex1 cells exhibit accelerated neurite extension upon NGF exposure, and have the capacity to regenerate neurites in the absence of NGF. In this study, we identify the repertoire of genes targeted by Nex1 to unravel the molecular mechanisms by which Nex1 promotes differentiation and regeneration. Our transcriptional analysis reveals that Nex1 modulates a wide spectrum of genes with diverse functions, many of them being key downstream regulators of the NGF pathway, and critical to neuritogenesis, such as microtubules, microtubule‐associated proteins (MAPs) and intermediate filaments. We also provide the first evidence that a basic helix‐loop‐helix (bHLH) protein stimulates the expression of the cyclin‐dependent kinase (CDK) inhibitors belonging to the INK4 family, which plays a role in promoting cell‐cycle arrest. Finally, we show a dramatic synergistic effect between Nex1 and cAMP, resulting in an impressive regeneration of an elaborate and dense neurite network. Thus, Nex1 has endowed the PC12‐Nex1 cells with a distinct combination of gene products that takes part in the complex regulation of neuritogenesis and regeneration.