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The ERK phosphorylation levels in the amygdala predict anxiety symptoms in humans and MEK/ERK inhibition dissociates innate and learned defensive behaviors in rats

Molecular Psychiatry volume 26, pages 7257–7269 (2021)Cite this article

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Abstract

We demonstrate that the rate of extracellular signal-related kinase phosphorylation (P-ERK1,2/Total-ERK1,2) in the amygdala is negatively and independently associated with anxiety symptoms in 23 consecutive patients with drug-resistant mesial temporal lobe epilepsy that was surgically treated. In naive Wistar rats, the P-ERK1,2/Total-ERK1,2 ratio in the amygdala correlates negatively with innate anxiety-related behavior on the elevated plus maze (n = 20) but positively with expression of defensive-learned behavior (i.e., freezing) on Pavlovian aversive (fear) conditioning (n = 29). The microinfusion of ERK1/2 inhibitor (FR180204, n = 8–13/group) or MEK inhibitor (U0126, n = 8–9/group) into the basolateral amygdala did not affect anxiety-related behavior but impaired the evocation (anticipation) of conditioned-defensive behavior (n = 9–11/group). In conclusion, the P-ERK1,2/Total-ERK1,2 ratio in the amygdala predicts anxiety in humans and the innate anxiety- and conditioned freezing behaviors in rats. However, the ERK1/2 in the basolateral AMY is only required for the expression of defensive-learned behavior. These results support a dissociate ERK-dependent mechanism in the amygdala between innate anxiety-like responses and the anticipation of learned-defensive behavior. These findings have implications for understanding highly prevalent psychiatric disorders related to the defensive circuit manifested by anxiety and fear.

Highlights

  • The P-ERK1,2/Total-ERK1,2 ratio in the amygdala (AMY) correlates negatively with anxiety symptoms in patients with mesial temporal lobe epilepsy.

  • The P-ERK1,2/Total-ERK1,2 in the amygdala correlates negatively with the anxiety-like behavior and positively with freezing-learned behavior in naive rats.

  • ERK1,2 in the basolateral amygdala is required for learned-defensive but not for the anxiety-like behavior expression in rats.

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Fig. 1: Pearson’s correlation between the neurochemical parameter levels according to the brain areas and the HADS anxiety scores (n = 23).
Fig. 2: The P-ERK/Total-ERK levels in the AMY correlates negatively with the anxiety-like behavior, but pharmacological BLA inhibition of MEK/ERK did not change its behavior in rats.
Fig. 3: The P-ERK/Total-ERK levels in the AMY correlates positively with freezing behavior and MEK/ERK activation into the BLA is required for learned-defensive rats.

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Acknowledgements

This work was supported by PRONEX Program (Programa de Núcleos de Excelência - NENASC Project) of FAPESC-CNPq-MS, Santa Catarina Brazil (process 56802/2010), MRC 271–05–0712 (ZAB), FAPESC-CONFAP – THE UK ACADEMIES – 2016 (ZAB and RW), and Brazilian National Council for Scientific and Technological Development (CNPq) Grant 408210/2018–4 (RW). We are grateful to the Laboratório Multiusuário de Estudos em Biologia at the Universidade Federal de Santa Catarina (LAMEB/UFSC) for providing its infrastructure for carrying out the western blotting quantification. CRC, CLC, and AAH were supported by scholarships from CAPES/PNPD. RBL, RDP, AL, KL, and RW are researchers from the CNPq.

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Authors and Affiliations

  1. Graduate Program in Neuroscience, Universidade Federal de Santa Catarina (UFSC), Florianópolis, SC, Brazil

    Cristiane Ribeiro de Carvalho, Leandra C. Constantino, Hiago Murilo de Melo, Rui Daniel Prediger, Rodrigo Bainy Leal & Roger Walz

  2. Center for Applied Neuroscience, Hospital Universitário (HU), UFSC, Florianópolis, SC, Brazil

    Cristiane Ribeiro de Carvalho, Leandra C. Constantino, Alexandre Ademar Hoeller, Hiago Murilo de Melo, Ricardo Guarnieri, Marcelo Neves Linhares, Rui Daniel Prediger, Katia Lin, Rodrigo Bainy Leal & Roger Walz

  3. Graduate Program in Biochemistry, UFSC, Florianópolis, SC, Brazil

    Mark William Lopes & Rodrigo Bainy Leal

  4. Medical Science Research Group - Health Investigation, Medicine Graduation Course, Centro Universitário para o Desenvolvimento do Alto Vale do Itajaí (UNIDAVI), Rio do Sul, SC, Brazil

    Mark William Lopes

  5. Graduate Program in Medical Sciences, UFSC, Florianópolis, SC, Brazil

    Alexandre Ademar Hoeller, Katia Lin & Roger Walz

  6. Neurosurgery Unit, Governador Celso Ramos Hospital, Florianópolis, SC, Brazil

    Ricardo Guarnieri & Marcelo Neves Linhares

  7. Center of Epilepsy Surgery of Santa Catarina (CEPESC), UFSC, Florianópolis, SC, Brazil

    Ricardo Guarnieri, Marcelo Neves Linhares, Katia Lin & Roger Walz

  8. Psychiatry Division, HU, UFSC, Florianópolis, Brazil

    Ricardo Guarnieri

  9. Neurosurgery Service, Department of Surgery, HU, UFSC, Florianópolis, SC, Brazil

    Marcelo Neves Linhares

  10. Centre for Synaptic Plasticity, School of Physiology, Pharmacology and Neuroscience, University of Bristol, Bristol, UK

    Zuner Assis Bortolotto

  11. Department of Pharmacology, UFSC, Florianópolis, SC, Brazil

    Rui Daniel Prediger

  12. LABOX, Departamento de Bioquímica, Centro de Ciências Biológicas, UFSC, Florianópolis, SC, Brazil

    Alexandra Latini

  13. Neurology Service, Department of Internal Medicine, UFSC, Florianópolis, SC, Brazil

    Katia Lin & Roger Walz

  14. Department of Psychiatry and Behavioral Sciences, State University of New York, Upstate Medical University, Syracuse, NY, USA

    Julio Licinio

  15. Biochemistry Department, UFSC, Florianópolis, SC, Brazil

    Rodrigo Bainy Leal

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  1. Cristiane Ribeiro de Carvalho
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Contributions

Principal investigator (PI): RW. Study design: CRC, RBL, and RW. Method development and data analysis: CRC, MWL, LCC, HMM, AAH, KL, RBL, and RW. Clinical data collection: RG, KL, and RW. Human brain tissue sampling: MNL and RW. Stereotaxic surgery and rat tissue sampling: CRC, HMM, LCC, and AAH. Biochemical assays of human and rat samples: MWL, LCC, AL, and RBL. Experimental and analytical supervision: CRC, AAH, RDP, RBL, and RW. Manuscript writing: CRC and RW. Data interpretation: CRC, LCC, AAH, RDP, AL, ZAB, JL, RBL, and RW. Manuscript edition and approval: all authors.

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Correspondence to Roger Walz.

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de Carvalho, C.R., Lopes, M.W., Constantino, L.C. et al. The ERK phosphorylation levels in the amygdala predict anxiety symptoms in humans and MEK/ERK inhibition dissociates innate and learned defensive behaviors in rats. Mol Psychiatry 26, 7257–7269 (2021). https://doi.org/10.1038/s41380-021-01203-0

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