Selective cleavage of ErbB4 by G-protein-coupled Gonadotropin-Releasing Hormone Receptor in Cultured Hypothalamic Neurons

• Published in: Journal of cellular physiology 2012-06, Vol.227 (6), p.2492-2501
• Main Authors: Higa-Nakamine, Sayomi, Maeda, Noriko, Toku, Seikichi, Yamamoto, Tomoko, Yingyuenyong, Matee, Kawahara, Masahiro, Yamamoto, Hideyuki
• Format: Article
• Language: English
• Subjects: ADAM Proteins - antagonists & inhibitors; ADAM Proteins - genetics; ADAM Proteins - metabolism; ADAM17 Protein; Animals; Cell Line; Down-Regulation; Enzyme Activation; Enzyme Inhibitors - pharmacology; Extracellular Signal-Regulated MAP Kinases - metabolism; Gonadotropin-Releasing Hormone - metabolism; Hypothalamus - drug effects; Hypothalamus - metabolism; Mice; Neuregulin-1 - metabolism; Neurons - drug effects; Neurons - metabolism; Phosphorylation; Protein Processing, Post-Translational; Receptor, Epidermal Growth Factor - antagonists & inhibitors; Receptor, Epidermal Growth Factor - genetics; Receptor, Epidermal Growth Factor - metabolism; Receptor, ErbB-4; Receptors, LHRH - metabolism; RNA Interference; Time Factors; Transfection
• ISSN: 0021-9541; 1097-4652
• DOI: 10.1002/jcp.22988

Gonadotropin‐releasing hormone (GnRH) is secreted from hypothalamic neurons (GnRH neurons). GnRH neurons have a GnRH receptor belonging to the G‐protein‐coupled receptors. The stimulation of this receptor activates extracellular signal‐regulated kinase (ERK). In the present study, we found that epidermal growth factor receptor (EGFR) and ErbB4 were expressed in immortalized GnRH neurons (GT1‐7 cells). AG1478, a relatively specific inhibitor of the ErbB family, and small interfering RNA (siRNA) for ErbB4 inhibited the GnRH‐induced activation of ERK in GT1‐7 cells, suggesting that EGFR and ErbB4 were necessary for the activation. In addition, GnRH induced the cleavage of ErbB4 and accumulation of an 80‐kDa fragment. After treatment of the cells with 50 nM GnRH for 5 min, about 80% of ErbB4 was cleaved. Biotinylation of cell surface proteins revealed that more than 70% of the cell surface ErbB4 was cleaved by GnRH treatment. A higher concentration and longer treatment were necessary for GnRH to induce ErbB4 cleavage than ERK activation. TAPI‐2, an inhibitor of tumor necrosis factor‐α‐converting enzyme (TACE), and siRNA for TACE inhibited the cleavage of ErbB4, suggesting that TACE was involved. After ErbB4 cleavage, the activation of ERK by neuregulin 1 was almost completely inhibited. These results suggest that the down‐regulation of ErbB4 expression is induced by G‐protein‐coupled receptor stimulation. J. Cell. Physiol. 227: 2492–2501, 2012. © 2011 Wiley Periodicals, Inc.