FK962 promotes neurite elongation and regeneration of cultured rat trigeminal ganglion cells: possible involvement of GDNF.
OBJECTIVE
Amputation of the trigeminal nerve leads to decreased corneal sensitivity and dry eye. Our previous study showed that the drug FK962 (N-[1-acetylpiperidin-4-yl]-4-fluorobenzamide) induced neurite elongation from trigeminal ganglion (TG) cells and accelerated recovery of corneal sensitivity in a rabbit model of in situ keratomileusis (LASIK) surgery. However, the molecular pathways leading to FK962-induced neurite elongation and regeneration are not well defined. Thus, the purposes of the present experiments were to determine if FK962 induces elongation and regeneration of cultured rat TG cells and to investigate the mechanism of FK962-induced neurite elongation.


METHODS
Mixed TG cells were cultured with or without FK962, and immunocytochemistry was used to detect stimulation of neurite elongation. Neurite regeneration was also tested in an in vitro model of neuronal ablation. ELISA was used to detect glial cell line-derived neurotrophic factor (GDNF) and somatostatin (SST) release, and mRNA expression was measured by qPCR. Antibody neutralization was used to determine the mechanism for FK962-induced neurite elongation/regeneration.


RESULTS
FK962 enhanced elongation and regeneration of neurites in TG neurons. GDNF treatment-induced neurite elongation and GDNF antibody significantly inhibited neurite elongation induced by GDNF and FK962. Nerve growth factor (NGF) treatment also induced neurite elongation, which was inhibited by NGF antibody, but NGF antibody did not inhibit FK962-induced neurite elongation.


CONCLUSIONS
Our data suggested that FK962 stimulated induction of GDNF from TG cells. GDNF may be a part of the signaling pathway for FK962-induced neurite elongation/regeneration in rat TG neurons.
Senju Laboratory of Ocular Sciences, Senju Pharmaceutical Co., Ltd., Kobe, Hyogo, Japan.
Yayoi
Y
Kishimoto
Chiho
C
Yabuta
Thomas R
TR
Shearer
Mitsuyoshi
M
Azuma
D000818:Animals; D001549:Benzamides / Q000494:pharmacology*; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000502:physiology*; D007150:Immunohistochemistry; D009416:Nerve Regeneration / Q000187:drug effects; D016501:Neurites / Q000502:physiology*; D010880:Piperidines / Q000494:pharmacology*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D060888:Real-Time Polymerase Chain Reaction; D012668:Trigeminal Ganglion / Q000502:physiology*
D001549:Benzamides; D051100:Glial Cell Line-Derived Neurotrophic Factor; C507659:N-(1-acetylpiperidin-4-yl)-4-fluorobenzamide; D010880:Piperidines; D012333:RNA, Messenger