Phosphoproteomics of the Dopamine Pathway Enables Discovery of Rap1 Activation as a Reward Signal In Vivo

Taku Nagai; Shinichi Nakamuta; Keisuke Kuroda; Sakura Nakauchi; Tomoki Nishioka; Tetsuya Takano; Xinjian Zhang; Daisuke Tsuboi; Yasuhiro Funahashi; Takashi Nakano; Junichiro Yoshimoto; Kenta Kobayashi; Motokazu Uchigashima; Masahiko Watanabe; Masami Miura; Akinori Nishi; Kazuto Kobayashi; Kiyofumi Yamada; Mutsuki Amano; Kozo Kaibuchi

doi:10.1016/j.neuron.2015.12.019

Phosphoproteomics of the Dopamine Pathway Enables Discovery of Rap1 Activation as a Reward Signal In Vivo

Neuron. 2016 Feb 3;89(3):550-65. doi: 10.1016/j.neuron.2015.12.019. Epub 2016 Jan 21.

Authors

Taku Nagai¹, Shinichi Nakamuta², Keisuke Kuroda², Sakura Nakauchi², Tomoki Nishioka², Tetsuya Takano², Xinjian Zhang³, Daisuke Tsuboi², Yasuhiro Funahashi², Takashi Nakano⁴, Junichiro Yoshimoto⁵, Kenta Kobayashi⁶, Motokazu Uchigashima⁷, Masahiko Watanabe⁷, Masami Miura⁸, Akinori Nishi⁹, Kazuto Kobayashi¹⁰, Kiyofumi Yamada¹, Mutsuki Amano², Kozo Kaibuchi¹¹

Affiliations

¹ Department of Neuropsychopharmacology and Hospital Pharmacy, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan; SRPBS, Japan Agency for Medical Research and Development, AMED 1-7-1 Otemachi, Chiyoda-ku, Tokyo 100-0004, Japan.
² Department of Cell Pharmacology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan; SRPBS, Japan Agency for Medical Research and Development, AMED 1-7-1 Otemachi, Chiyoda-ku, Tokyo 100-0004, Japan.
³ Department of Cell Pharmacology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan.
⁴ Neurobiology Research Unit, Okinawa Institute of Science and Technology Graduate University, Onna, Okinawa 904-0495, Japan; SRPBS, Japan Agency for Medical Research and Development, AMED 1-7-1 Otemachi, Chiyoda-ku, Tokyo 100-0004, Japan.
⁵ Neural Computation Unit, Okinawa Institute of Science and Technology Graduate University, Onna, Okinawa 904-0495, Japan; SRPBS, Japan Agency for Medical Research and Development, AMED 1-7-1 Otemachi, Chiyoda-ku, Tokyo 100-0004, Japan.
⁶ Section of Viral Vector Development, National Institute for Physiological Sciences, 38 Nishigonaka Myodaiji, Okazaki 444-8585, Japan.
⁷ Department of Anatomy, Hokkaido University Graduate School of Medicine, Kita 15, Nishi 7, Kita-ku, Sapporo 060-8638, Japan.
⁸ Neuropathophysiology Research Group, Tokyo Metropolitan Institute of Gerontology, 35-2 Sakae-cho, Itabashi-ku, Tokyo 173-0015, Japan.
⁹ Department of Pharmacology, School of Medicine, Kurume University, 67 Asahi-machi, Kurume 830-0011, Japan; SRPBS, Japan Agency for Medical Research and Development, AMED 1-7-1 Otemachi, Chiyoda-ku, Tokyo 100-0004, Japan.
¹⁰ Department of Molecular Genetics, Institute of Biomedical Sciences, School of Medicine, Fukushima Medical University, 1 Hikariga-oka, Fukushima 960-1295, Japan.
¹¹ Department of Cell Pharmacology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan; SRPBS, Japan Agency for Medical Research and Development, AMED 1-7-1 Otemachi, Chiyoda-ku, Tokyo 100-0004, Japan. Electronic address: kaibuchi@med.nagoya-u.ac.jp.

PMID: 26804993
DOI: 10.1016/j.neuron.2015.12.019

Abstract

Dopamine (DA) type 1 receptor (D1R) signaling in the striatum presumably regulates neuronal excitability and reward-related behaviors through PKA. However, whether and how D1Rs and PKA regulate neuronal excitability and behavior remain largely unknown. Here, we developed a phosphoproteomic analysis method to identify known and novel PKA substrates downstream of the D1R and obtained more than 100 candidate substrates, including Rap1 GEF (Rasgrp2). We found that PKA phosphorylation of Rasgrp2 activated its guanine nucleotide-exchange activity on Rap1. Cocaine exposure activated Rap1 in the nucleus accumbens in mice. The expression of constitutively active PKA or Rap1 in accumbal D1R-expressing medium spiny neurons (D1R-MSNs) enhanced neuronal firing rates and behavioral responses to cocaine exposure through MAPK. Knockout of Rap1 in the accumbal D1R-MSNs was sufficient to decrease these phenotypes. These findings demonstrate a novel DA-PKA-Rap1-MAPK intracellular signaling mechanism in D1R-MSNs that increases neuronal excitability to enhance reward-related behaviors.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Action Potentials / physiology
Animals
Benzazepines / pharmacology
Cocaine / pharmacology
Colforsin / pharmacology
Corpus Striatum / drug effects
Corpus Striatum / metabolism
Cyclic AMP-Dependent Protein Kinases / metabolism
Dopamine / metabolism*
Dopamine / pharmacology
Enzyme Activation
Extracellular Signal-Regulated MAP Kinases / metabolism
Extracellular Signal-Regulated MAP Kinases / physiology
Guanine Nucleotide Exchange Factors / metabolism
Mice
Mice, Knockout
Neurons / metabolism
Neurons / physiology
Nucleus Accumbens / metabolism
Phosphoproteins / metabolism*
Phosphorylation / drug effects
Proteome / metabolism*
Proteomics*
Receptors, Dopamine D1 / metabolism*
Reward*
Signal Transduction* / drug effects
rap1 GTP-Binding Proteins / genetics
rap1 GTP-Binding Proteins / metabolism*

Substances

Benzazepines
Guanine Nucleotide Exchange Factors
Phosphoproteins
Proteome
Rasgrp2 protein, mouse
Receptors, Dopamine D1
Colforsin
SK&F 81297
Cyclic AMP-Dependent Protein Kinases
Extracellular Signal-Regulated MAP Kinases
rap1 GTP-Binding Proteins
Cocaine
Dopamine