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Updated: Feb 11, 2026

Protocol for Studying Extinction of Conditioned Fear in Naturally Cycling Female Rats
Published on: February 23, 2015
Effects of estrus cycle stage on defensive behavior in female Long-Evans hooded rats
Nathan S Pentkowski1, Yoav Litvin1, D Caroline Blanchard2
1Department of Psychology, University of Hawaii at Manoa, 2530 Dole Street, Sakamaki C 400, Honolulu, HI 96822, USA.
Abstract:
This study investigated the influence of the estrus cycle in mediating cat odor-induced unconditioned and conditioned defensive behaviors in female Long-Evans hooded rats. Unconditioned defensive behaviors were assessed during predatory cue exposure; conditioned defensive behaviors were examined 24 h after threat exposure. Estrus phases were determined by microscopic examination of vaginal smears within 10 min of completing the behavioral tests. Compared to no-odor controls, female rats exposed to cat odor exhibited both unconditioned and conditioned defensive behaviors, including elevated levels of freezing, risk assessment and avoidance. Rats in proestrus and estrus exhibited reduced levels of defensive behavior during the unconditioned test trial compared to subjects in diestrus and metestrus. Specifically, estrus stages characterized by high levels of circulating estrogens and progesterone were associated with reduced immobility (i.e. freezing) and enhanced active defense (i.e. risk assessment), profiles that may enable mate seeking and subsequent reproduction in potentially dangerous or novel environments. These results suggest a specific role for ovarian hormone fluctuations in mediating unconditioned fear- and anxiety-like defensive behaviors during exposure to predatory odors.
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