title: Tricyclic Antidepressant Amitriptyline-induced Glial Cell Line-derived Neurotrophic Factor Production Involves Pertussis Toxin-sensitive Gαi/o Activation in Astroglial Cells.
abstract: Further elaborating the mechanism of antidepressants, beyond modulation of monoaminergic neurotransmission, this study sought to elucidate the mechanism of amitriptyline-induced production of glial cell line-derived neurotrophic factor (GDNF) in astroglial cells. Previous studies demonstrated that an amitriptyline-evoked matrix metalloproteinase (MMP)/FGF receptor (FGFR)/FGFR substrate 2α (FRS2α)/ERK cascade is crucial for GDNF production, but how amitriptyline triggers this cascade remains unknown. MMP is activated by intracellular mediators such as G proteins, and this study sought to clarify the involvement of G protein signaling in amitriptyline-evoked GDNF production in rat C6 astroglial cells (C6 cells), primary cultured rat astrocytes, and normal human astrocytes. Amitriptyline-evoked GDNF mRNA expression and release were inhibited by pertussis toxin (PTX), a Gα(i/o) inhibitor, but not by NF449, a Gα(s) inhibitor, or YM-254890, a Gαq inhibitor. The activation of the GDNF production cascade (FGFR/FRS2α/ERK) was also inhibited by PTX. Deletion of Gα(ο1) and Gα(i3) by RNAi demonstrated that these G proteins play important roles in amitriptyline signaling. G protein activation was directly analyzed by electrical impedance-based biosensors (CellKey(TM) assay), using a label-free (without use of fluorescent proteins/probes or radioisotopes) and real time approach. Amitriptyline increased impedance, indicating Gα(i/o) activation that was suppressed by PTX treatment. The impedance evoked by amitriptyline was not affected by inhibitors of the GDNF production cascade. Furthermore, FGF2 treatment did not elicit any effect on impedance, indicating that amitriptyline targets PTX-sensitive Gα(i/o) upstream of the MMP/FGFR/FRS2α/ERK cascade. These results suggest novel targeting for the development of antidepressants.
authors: From the Department of Pharmacology, Hiroshima University Graduate School of Biomedical Sciences, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553, hisaokak@hiroshima-u.ac.jp.
authors: Kazue
authors: K
authors: Hisaoka-Nakashima
authors: the Division of Cancer Pathophysiology, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045, and.
authors: Kanako
authors: K
authors: Miyano
authors: From the Department of Pharmacology, Hiroshima University Graduate School of Biomedical Sciences, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553.
authors: Chie
authors: C
authors: Matsumoto
authors: From the Department of Pharmacology, Hiroshima University Graduate School of Biomedical Sciences, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553.
authors: Naoto
authors: N
authors: Kajitani
authors: From the Department of Pharmacology, Hiroshima University Graduate School of Biomedical Sciences, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553, the Division of Psychiatry and Neuroscience, Institute for Clinical Research, and.
authors: Hiromi
authors: H
authors: Abe
authors: the Division of Psychiatry and Neuroscience, Institute for Clinical Research, and.
authors: Mami
authors: M
authors: Okada-Tsuchioka
authors: the Division of Cancer Pathophysiology, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045, and.
authors: Akinobu
authors: A
authors: Yokoyama
authors: the Division of Cancer Pathophysiology, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045, and.
authors: Yasuhito
authors: Y
authors: Uezono
authors: From the Department of Pharmacology, Hiroshima University Graduate School of Biomedical Sciences, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553.
authors: Norimitsu
authors: N
authors: Morioka
authors: From the Department of Pharmacology, Hiroshima University Graduate School of Biomedical Sciences, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553.
authors: Yoshihiro
authors: Y
authors: Nakata
authors: the Division of Psychiatry and Neuroscience, Institute for Clinical Research, and Department of Psychiatry, National Hospital Organization Kure Medical Center and Chugoku Cancer Center, 3-1 Aoyama, Kure 737-0023, Japan mtakebayashi@kure-nh.go.jp.
authors: Minoru
authors: M
authors: Takebayashi
mesh_terms: D000639:Amitriptyline / Q000494:pharmacology*; D000818:Animals; D000929:Antidepressive Agents, Tricyclic / Q000494:pharmacology*; D001253:Astrocytes / Q000166:cytology / Q000378:metabolism*; D015374:Biosensing Techniques; D002460:Cell Line; D003865:Major Depressive Disorder / Q000188:drug therapy; D017097:Electric Impedance; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D019206:GTP-Binding Protein alpha Subunits, Gi-Go / Q000378:metabolism*; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D008297:Male; D037342:Pertussis Toxin / Q000737:chemistry*; D010766:Phosphorylation; D012313:RNA / Q000378:metabolism; D034622:RNA Interference; D051381:Rats; D011957:Receptors, Opioid / Q000378:metabolism; D015398:Signal Transduction
chemical_list: D000929:Antidepressive Agents, Tricyclic; C495039:GDNF protein, human; C495041:Gdnf protein, rat; D051100:Glial Cell Line-Derived Neurotrophic Factor; D011957:Receptors, Opioid; D000639:Amitriptyline; D012313:RNA; D037342:Pertussis Toxin; D048049:Extracellular Signal-Regulated MAP Kinases; D019206:GTP-Binding Protein alpha Subunits, Gi-Go
keywords: Cellkey assay; G protein; antidepressant; astrocyte; biosensor; depression; glial cell