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CAPON-nNOS coupling can serve as a target for developing new anxiolytics

Nature Medicine volume 20, pages 1050–1054 (2014)Cite this article

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Abstract

Anxiety disorders are highly prevalent psychiatric diseases1,2. There is need for a deeper understanding of anxiety control mechanisms in the mammalian brain and for development of new anxiolytic agents. Here we report that the coupling between neuronal nitric oxide synthase (nNOS) and its carboxy-terminal PDZ ligand (CAPON) can serve as a target for developing new anxiolytic agents. Augmenting nNOS-CAPON interaction in the hippocampus of mice by overexpressing full-length CAPON gave rise to anxiogenic-like behaviors, whereas dissociating CAPON from nNOS by overexpressing CAPON-125C or CAPON-20C (the C-terminal 125 or 20 amino acids of CAPON) or delivering Tat-CAPON-12C (a peptide comprising Tat and the 12 C-terminal amino acids of CAPON) in the hippocampus of mice produced anxiolytic-like effects. Mice subjected to chronic mild stress (CMS) displayed a substantial increase in nNOS-CAPON coupling in the hippocampus and a consequent anxiogenic-like phenotype. Disrupting nNOS-CAPON coupling reversed the CMS-induced anxiogenic-like behaviors. Moreover, small-molecule blockers of nNOS-CAPON binding rapidly produced anxiolytic-like effects. Dexamethasone-induced ras protein 1 (Dexras1)–extracellular signal–regulated kinase (ERK) signaling was involved in the behavioral effects of nNOS-CAPON association. Thus, nNOS-CAPON association contributes to the modulation of anxiety-related behaviors via regulating Dexras1-ERK signaling and can serve as a target for developing potential anxiolytics.

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Figure 1: Augmenting nNOS-CAPON interaction causes anxiogenic-like behaviors.
Figure 2: Uncoupling nNOS and CAPON produces anxiolytic-like effects.
Figure 3: Blocking nNOS-CAPON association reverses stress-induced behavioral modifications.
Figure 4: nNOS-CAPON blockers produce anxiolytic-like effects via Dexras1-ERK signaling.

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Acknowledgements

This work was supported by grants from the National Natural Science Foundation of China (91232304, 81222016, 81030023 and 21303086), the National Basic Research Program of China (973 Program) (2011CB504404) and the Natural Science Foundation of Jiangsu Province–Special Program grant BK2011029 and Distinguished Young Scientist Fund BK20130040, and by the Collaborative Innovation Center For Cardiovascular Disease Translational Medicine. We thank X.D. Qian, H.Y. Ni, F.Y. Zhang, Y. Hu, Y. Tang and Z. Zhu for technical assistance.

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Author notes

  1. Wei Lu

    Present address: Present address: Key Laboratory of Developmental Genes and Human Disease, Ministry of Education, Institute of Life Sciences, Southeast University, Nanjing, China.,

  2. Li-Juan Zhu and Ting-You Li: These authors contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Cerebrovascular Disease, Key Laboratory of Cardiovascular Disease and Molecular Intervention, Nanjing Medical University, Nanjing, China

    Li-Juan Zhu, Chun-Xia Luo, Lei Chang, Yu-Hui Lin, Hai-Hui Zhou, Chen Chen, Li-Yan Gao, Hai-Yin Wu & Dong-Ya Zhu

  2. Department of Pharmacology, Nanjing Medical University, Nanjing, China

    Li-Juan Zhu, Chun-Xia Luo, Yu-Hui Lin, Hai-Hui Zhou, Chen Chen, Yu Zhang, Li-Yan Gao, Qi-Gang Zhou, Jing Zhang, Hai-Yin Wu & Dong-Ya Zhu

  3. Department of Medicinal Chemistry, Nanjing Medical University, Nanjing, China

    Ting-You Li, Nan Jiang, Lei Chang & Yu Ma

  4. Department of Neurobiology, Nanjing Medical University, Nanjing, China

    Wei Lu

  5. Department of Chemistry, Nanjing Medical University, Nanjing, China

    Qin Hu

  6. Center of Drug Metabolism & Pharmacokinetics, Jiangsu Simcere Pharmaceutical, Nanjing, China

    Xiao-Ling Hu

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Contributions

L.-J.Z. and C.C. performed the cell culture studies, coimmunoprecipitation, western blotting, surgical preparation and animal behavioral examinations. T.-Y.L. and L.C. contributed the design and synthesis of small-molecule compounds. Y.Z. and H.-H.Z. contributed recombinant lentivirus production and Tat-CAPON-12C preparation. N.J. performed molecular dynamics simulations, interaction energy calculations and force field parameterization for ZLc-002. C.-X.L. contributed to the design of the study. W.L. performed electrophysiological experiments. L.-Y.G., Y.-H.L., Y.M., Q.-G.Z., J.Z. and H.-Y.W. participated in the study. Q.H. and X.-L.H. analyzed concentrations of ZLc-002 and its metabolites. D.-Y.Z. initiated the project, designed the study and wrote the paper. All authors contributed to data analysis.

Corresponding authors

Correspondence to Chun-Xia Luo or Dong-Ya Zhu.

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Zhu, LJ., Li, TY., Luo, CX. et al. CAPON-nNOS coupling can serve as a target for developing new anxiolytics. Nat Med 20, 1050–1054 (2014). https://doi.org/10.1038/nm.3644

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