Intraneuronal localization of Nogo‐A in the rat

Nogo‐A is known to be a myelin‐associated protein with strong inhibitory effect on neurite outgrowth and has been considered one of the major factors that hinder fiber regeneration in the central nervous system. Recent studies have demonstrated widespread occurrence of nogo‐A mRNA and Nogo‐A protein in neurons. Our concurrent immunohistochemical study substantiated the widespread distribution of neuronal Nogo‐A. The present study was thus focused on its intraneuronal distribution in the central nervous system, using Western blotting, immunohistochemical, and immunogold electron microscopic techniques. Western blotting of the nucleus, cytoplasm, and membrane subcellular fractions of the cerebellum and spinal cord tissues demonstrated that all three fractions contained Nogo‐A. Nogo‐A immunoreactivity could be identified under confocal microscope in the nucleus, perikayon, and proximal dendrite and along the cell membrane. Under the electron microscope, the perikaryonal Nogo‐A immunogold particles were mainly distributed at polyribosomes and rough endoplasmic reticulum, suggesting its relationship with translation process. The immunogold particles could also be found beneath or on the plasma membrane. In the nucleus, the Nogo‐A immunogold particles were found to be localized at the chromatins of the nucleus, indicating its possible involvement in gene transcription. The presence of Nogo‐A in the nucleus was further supported by transfection of COS‐7L cells with nogo‐A. This study provides the first immunocytochemical evidence for intraneuronal distribution of Nogo‐A. Apparently, the significance of Nogo‐A in the central nervous system is far more complex than what has been envisioned. J. Comp. Neurol. 458:1–10, 2003. © 2003 Wiley‐Liss, Inc.