Distinct contributions of median raphe nucleus to contextual fear conditioning and fear-potentiated startle.

J Landeira-Fernandez,R C B Silva,V Avanzi,A P M Cruz,M L Brandão

doi:10.1155/np.2002.233

Abstract

Ascending 5-HT projections from the median raphe nucleus (MRN), probably to the hippocampus, are implicated in the acquisition of contextual fear (background stimuli), as assessed by freezing behavior. Foreground cues like light, used as a conditioned stimulus (CS) in classical fear conditioning, also cause freezing through thalamic transmission to the amygdala. As the MRN projects to the hippocampus and amygdala, the role of this raphe nucleus in fear conditioning to explicit cues remains to be explained. Here we analyzed the behavior of rats with MRN electrolytic lesions in a contextual conditioning situation and in a fear-potentiated startle procedure. The animals received MRN electrolytic lesions either before or on the day after two consecutive training sessions in which they were submitted to 10 conditioning trials, each in an experimental chamber (same context) where they. received foot-shocks (0.6 mA, 1 sec) paired to a 4-sec light CS. Seven to ten days later, the animals were submitted to testing sessions for assessing conditioned fear when they were placed for five shocks, and the duration of contextual freezing was recorded. The animals were then submitted to a fear-potentiated startle in response to a 4-sec light-CS, followed by white noise (100 dB, 50 ms). Control rats (sham) tested in the same context showed more freezing than did rats with pre- or post-training MRN lesions. Startle was clearly potentiated in the presence of light CS in the sham-lesioned animals. Whereas pretraining lesions reduced both freezing and fear-potentiated startle, the post-training lesions reduced only freezing to context, without changing the fear-potentiated startle. In a second experiment, neurotoxic lesions of the MRN with local injections of N-methyl-D-aspartate or the activation of 5-HT1A somatodendritic auto-receptors of the MRN by microinjections of the 5-HT1A receptor agonist 8-hydroxy- 2-(di-n-propylamino)tetralin (8-OH-DPAT) before the training sessions also reduced the amount of freezing and the fear-potentiated startle. Freezing is a prominent response of contextual fear conditioning, but does not seem to be crucial for the enhancement of the startle reflex by explicit aversive cues. As fear-potentiated startle may be produced in posttraining lesioned rats that are unable to freeze to fear contextual stimuli, dissociable systems seem to be recruited in each condition. Thus, contextual fear and fear-potentiated startle are conveyed by distinct 5-HT-mediated circuits of the MRN.

Highlights

Fear conditioning to context can be reliably evaluated by the amount of freezing behavior that animals display when they return to the context in which they received footshock and fear-potentiated startle by the increase of the startle reflex in the presence of an explicit cue paired with shock (Brown et al, 1951; Kim & Fanselow, 1992; Davis, 1992; Philips & LeDoux, 1992, 1994; McNish et al, 1997)
That electrolytic lesion of the median raphe nucleus (MRN) inhibited the freezing behavior to background cues in the chamber where the animals had previously experienced footshock provides evidence for an involvement of the MRN in contextual conditioned fear
The results cannot be attributed to changes in footshock sensitivity, as it has already been shown that lesions of the MRN did not affect freezing behavior immediately after shock presentations even though the same rats showed a deficit in freezing measured 48 h later, without footshock (Melik et al, 2000; Avanzi & Brandgo, 2001)

Summary

Introduction

Fear conditioning to context can be reliably evaluated by the amount of freezing behavior that animals display when they return to the context in which they received footshock and fear-potentiated startle by the increase of the startle reflex in the presence of an explicit cue paired with shock (Brown et al, 1951; Kim & Fanselow, 1992; Davis, 1992; Philips & LeDoux, 1992, 1994; McNish et al, 1997). A number of studies have demonstrated that electrolytic or neurotoxic lesions of the median raphe nucleus (MRN) severely disturb the performance of rats in a variety of behavioral tasks. Such a lesion may produce heightened motor activity and increased approach, aberrant hypersexuality, increased feeding behavior, and reduced escape and avoidance responses (Kiser & Lebovitz, 1975; Srebro & Lorens, 1975; Geyer et al, 1976; Tye et al, 1977; Hillegaart & Hjort, 1989; Hillegaart, 1990; Andrade & Graeff, 2001). Electrical stimulation of this nucleus causes reduced motor output and several autonomic signs, such as micturition and defecation (Graeff & Silveira-Filho, 1978)

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