The rostrodorsal periaqueductal gray influences both innate fear responses and acquisition of fear memory in animals exposed to a live predator

  • Rodrigo de Andrade Rufino
  • Sandra Regina Mota-Ortiz
  • Miguel Antonio Xavier De Lima
  • Marcus Vinicius C. Baldo
  • Newton Sabino CanterasEmail author
Original Article


A few studies have evaluated the behavioral roles of the periaqueductal gray (PAG) in animals facing ethologically relevant threats. Exposure to a live cat induces striking activation in the rostrodorsal and caudal ventral PAG. In the present investigation, we first showed that cytotoxic lesions of the rostrodorsal and caudal ventral PAG had similar effects on innate fear responses during cat exposure, practically abolishing freezing and increasing risk assessment responses. Conversely, rostrodorsal PAG lesions but not caudal ventral lesions disrupted learned contextual fear responses to cat exposure. Next, we examined how muscimol inactivation of the rostrodorsal PAG at different times (i.e., during, immediately after and 20 min after cat exposure) influences learned contextual fear responses, and we found that inactivation of the rostrodorsal PAG during or immediately after cat exposure but not 20 min later impaired contextual fear learning. Thus, suggesting that the rostrodorsal PAG is involved in the acquisition, but not the consolidation, of contextual fear memory to predatory threat. Notably, the dosolateral PAG contains a distinct population of neurons containing the neuronal nitric oxide synthase (nNOS) enzyme, and in the last experiment, we investigated how nitric oxide released in rostrodorsal PAG influences contextual fear memory processing. Accordingly, injection of a selective nNOS inhibitor into the rostrodorsal PAG immediately after cat exposure disrupted learned contextual responses. Overall, the present findings suggest that the acquisition of contextual fear learning is influenced by an optimum level of dorsal PAG activation, which extends from during to shortly after predator exposure and depends on local NO release.


Defensive behavior Fear memory Innate fear Nitric oxide 



This research was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)—Research Grants #2014/05432-9 (awarded to N.S.C) and #2009/53390-5 (awarded to S.R.M.O.). R.A.R. and M.A.X.L. were supported by FAPESP fellowships (R.A.R. #2014/02540-5/M.A.X.L. #2016/10389-0).

Compliance with ethical standards

Conflict of interest

There is no conflict of interest.

Research involving human participants and/or animals

Experiments were carried out in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals (NIH Publications No. 80-23, 1996). All experimental procedures had been previously approved by the Committee on The Care and Use of Laboratory Animals of the Institute of Biomedical Sciences, University of São Paulo, Brazil (Protocol number 085/2012). In the present study, the experiments were planned to minimize the number of animals used and their suffering. In addition, all surgical procedures were performed under deep anesthesia, and analgesic and antibiotic medications were given postoperatively.

Supplementary material

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Supplementary material 1 (DOC 9805 KB)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Rodrigo de Andrade Rufino
    • 1
  • Sandra Regina Mota-Ortiz
    • 1
  • Miguel Antonio Xavier De Lima
    • 1
  • Marcus Vinicius C. Baldo
    • 2
  • Newton Sabino Canteras
    • 1
    • 3
    Email author
  1. 1.Department of Anatomy, Institute of Biomedical SciencesUniversity of São PauloSão PauloBrazil
  2. 2.Department of Physiology and Biophysics, Institute of Biomedical SciencesUniversity of São PauloSão PauloBrazil
  3. 3.Department of Anatomy, Institute of Biomedical SciencesUniversity of Sao PauloSão PauloBrazil

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