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. 2015 Sep 9:9:321.
doi: 10.3389/fnins.2015.00321. eCollection 2015.

Temporary inactivation of the anterior part of the bed nucleus of the stria terminalis blocks alarm pheromone-induced defensive behavior in rats

Tino Breitfeld  1 , Johann E A Bruning  1 , Hideaki Inagaki  2 , Yukari Takeuchi  3 , Yasushi Kiyokawa  3 , Markus Fendt  4
Affiliations

Temporary inactivation of the anterior part of the bed nucleus of the stria terminalis blocks alarm pheromone-induced defensive behavior in rats

Tino Breitfeld et al. Front Neurosci. .

Abstract

Rats emit an alarm pheromone in threatening situations. Exposure of rats to this alarm pheromone induces defensive behaviors, such as head out behavior, and increases c-Fos expression in brain areas involved in the mediation of defensive behaviors. One of these brain areas is the anterior bed nucleus of the stria terminalis (aBNST). The goal of the present study was to investigate if pharmacological inactivation of the aBNST by local microinjections of the GABAA receptor-agonist muscimol modulates alarm pheromone-induced defensive behaviors. We first established the behavioral paradigm of alarm pheromone-induced defensive behaviors in Sprague-Dawley rats in our laboratory. In a second experiment, we inactivated the aBNST, then exposed rats to one of four different odors (neck odor, female urine, alarm pheromone, fox urine) and tested the effects of the aBNST inactivation on the behavior in response to these odors. Our data show that temporary inactivation of the aBNST blocked head out behavior in response to the alarm pheromone. This indicates that the aBNST plays an important role in the mediation of the alarm pheromone-induced defensive behavior in rats.

Keywords: BNST; alarm pheromone; anxiety; fear; muscimol; odor-induced anxiety; rats; risk assessment behavior.

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Figures

Figure 1
Figure 1
Effects of the alarm pheromone on the behavior of Sprague-Dawley rats. Rats were exposed to either the alarm pheromone (alarm) or water. Data are expressed as means ± SEMs. *p < 0.05, paired t-tests.
Figure 2
Figure 2
Injection sites into the aBNST. (A) Reconstruction of the different injection sites of saline or muscimol into the aBNST. The coronal sections were taken from the atlas of Paxinos and Watson (2014). Numbers indicate distance from bregma in mm. aBNST, anterior bed nucleus of the stria terminalis; MnPo, median preoptic nucleus; MS, medial septal nucleus; CPu, caudate putamen; LPO, lateral preoptic area; VP, ventral pallidum; f, fornix. (B) Photomicrographs with a representative example of aBNST injection sites.
Figure 3
Figure 3
Locomotor activity during the acclimation phase. Rats were exposed to neck odor (neck), female urine (female), alarm pheromone (alarm), or fox urine (fox). Neither intra-aBNST injections of muscimol nor the type of odor samples affected distance traveled. Data are expressed as means ± SEMs.
Figure 4
Figure 4
Effects of odors and aBNST inactivation on defensive behavior. Rats were exposed to neck odor (neck), female urine (female), alarm pheromone (alarm), or fox urine (fox). (A) Head out behavior, (B) time spent in the center, in percentage of the time spent outside. Data are expressed as means ± SEMs. *p < 0.05, ++p < 0.01, comparison with saline/alarm pheromone.
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