title: Valproic Acid Influences MTNR1A Intracellular Trafficking and Signaling in a β-Arrestin 2-Dependent Manner.
abstract: Valproate exposure is associated with increased risks of autism spectrum disorder. To date, the mechanistic details of disturbance of melatonin receptor subtype 1 (MTNR1A) internalization upon valproate exposure remain elusive. By expressing epitope-tagged receptors (MTNR1A-EGFP) in HEK-293 and Neuro-2a cells, we recorded the dynamic changes of MTNR1A intracellular trafficking after melatonin treatment. Using time-lapse confocal microscopy, we showed in living cells that valproic acid interfered with the internalization kinetics of MTNR1A in the presence of melatonin. This attenuating effect was associated with a decrease in the phosphorylation of PKA (Thr197) and ERK (Thr202/Tyr204). VPA treatment did not alter the whole-cell currents of cells with or without melatonin. Furthermore, fluorescence resonance energy transfer imaging data demonstrated that valproic acid reduced the melatonin-initiated association between YFP-labeled β-arrestin 2 and CFP-labeled MTNR1A. Together, we suggest that valproic acid influences MTNR1A intracellular trafficking and signaling in a β-arrestin 2-dependent manner.
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, China.
authors.forename: Ling-Juan
authors.initials: LJ
authors.lastname: Hong
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, China.
authors.forename: Quan
authors.initials: Q
authors.lastname: Jiang
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, China.|Department of Pharmacy, Hangzhou No.7 People's Hospital, Hangzhou, China.
authors.forename: Sen
authors.initials: S
authors.lastname: Long
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, China.
authors.forename: Huan
authors.initials: H
authors.lastname: Wang
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, China.|Department of Pharmacy, Zhejiang Xiaoshan Hospital, Hangzhou, Zhejiang, China.
authors.forename: Ling-di
authors.initials: LD
authors.lastname: Zhang
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Tian
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, China.
authors.forename: Cheng-Kun
authors.initials: CK
authors.lastname: Wang
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, China.
authors.forename: Jing-Jing
authors.initials: JJ
authors.lastname: Cao
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, China.
authors.forename: Rong-Rong
authors.initials: RR
authors.lastname: Tao
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, China.
authors.forename: Ji-Yun
authors.initials: JY
authors.lastname: Huang
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, China.
authors.forename: Mei-Hua
authors.initials: MH
authors.lastname: Liao
authors.affiliation: School of Medicine, Zhejiang University City College, Hangzhou, Zhejiang, China.
authors.forename: Ying-Mei
authors.initials: YM
authors.lastname: Lu
authors.affiliation: Department of Pharmacology, Graduate School of Pharmaceutical Sciences, Tohoku University, Sendai, Japan.
authors.forename: Kohji
authors.initials: K
authors.lastname: Fukunaga
authors.affiliation: Institute of Biochemistry, College Life Science, Zhejiang University, Hangzhou, China. znm2000@yahoo.com.
authors.forename: Nai-Ming
authors.initials: NM
authors.lastname: Zhou
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, China. changhuahan@zju.edu.cn.
authors.forename: Feng
authors.initials: F
authors.lastname: Han
mesh_terms: D000818:Animals; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D004705:Endocytosis / Q000187:drug effects; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer; D057809:HEK293 Cells; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism*; D008550:Melatonin / Q000494:pharmacology; D051379:Mice; D010766:Phosphorylation / Q000187:drug effects; D011485:Protein Binding / Q000187:drug effects; D021381:Protein Transport / Q000187:drug effects; D044122:Receptor, Melatonin, MT1 / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects*; D014635:Valproic Acid / Q000494:pharmacology*; D000071557:beta-Arrestins / Q000378:metabolism*; D020691:rab GTP-Binding Proteins / Q000378:metabolism
chemical_list: D044122:Receptor, Melatonin, MT1; D000071557:beta-Arrestins; D014635:Valproic Acid; D017868:Cyclic AMP-Dependent Protein Kinases; D048049:Extracellular Signal-Regulated MAP Kinases; D020691:rab GTP-Binding Proteins; D008550:Melatonin
keywords: FRET; Internalization; Melatonin; Melatonin receptor subtype 1; Valproic acid