Bruce S. Kapp

Amygdala central nucleus lesions: Effect on heart rate conditioning in the rabbit

Abstract The present study was designed as an initial step in determining the specific anatomical systems of the amygdala which may contribute to the expression of conditioned heart rate responding during aversive Pavlovian conditioning in the New Zealand rabbit. Animals receiving either small or large radio-frequency lesions of the central nucleus of the amygdala demonstrated a significant attenuation of the conditioned bradycardia response to the conditioned stimulus when compared to that demonstrated by control animals. No significant effects of the lesions on baseline heart rate or on the heart rate orienting response to the conditioned stimulus were observed. Central nucleus lesions also produced a significant effect on the unconditioned heart rate response to the unconditioned stimulus manifested in an increased duration and prolonged habituation of the unconditioned response. The results are consistent with anatomical and physiological evidence suggesting the involvement of the central nucleus in the expression of emotional responses including accompanying cardiovascular alterations.

Electrophysiological characteristics of amygdaloid central nucleus neurons during Pavlovian fear conditioning in the rabbit

Recent evidence suggests that the amygdaloid central nucleus (ACE) may contribute importantly to cardiovascular adjustments in response to the presentation of conditioned emotional stimuli, possibly via direct ACE projections to cardiovascular regulatory nuclei in the medulla. The present experiment was conducted to obtain additional data relevant to this suggestion. Extracellular single-unit recordings were obtained from 85 histologically-verified ACE neurons during Pavlovian differentially conditioned heart-rate responding in rabbits. Conditioning involved pairing one tone (CS+), but not a second tone (CS-), with paraorbital shock. Those ACE neurons which project to the lower brainstem were identified by their antidromic responses to stimulation of a mesencephalic region through which descending ACE projections course. Under these conditions it was possible to classify ACE neurons as conforming to one of 6 general categories based on their spontaneous activity and conditioned response characteristics. In addition, it was determined that: (1) the electrophysiological characteristics of many ACE neurons were differentially altered in response to presentations of the CS+ versus the CS-; (2) the responses of many ACE neurons to presentations of the CS+ were correlated with the magnitudes of concomitant conditioned alterations in heart rate; and (3) the activity of antidromically-identified ACE neurons which project to the lower brainstem was decreased in response to presentations of each CS. These data provide additional support for the notion that the ACE contributes to cardiovascular regulation during the presentation of emotionally-arousing stimuli.

Cardiovascular responses elicited by electrical stimulation of the amygdala central nucleus in the rabbit

Recent behavioral and anatomical evidence suggests the involvement of the amygdala central nucleus projection to medullary cardioregulatory nuclei in the expression of conditioned bradycardia during aversive Pavlovian conditioning in the rabbit 6,7,11,15. The present study sought to determine the extent to which electrical stimulation of the central nucleus produces bradycardia in the rabbit, and the extent to which any bradycardia elicited varies with stimulation at sites within as opposed to adjacent to the medial component of the central nucleus, the component from which the projection to cardioregulatory nuclei originates. Over 900 sites in 30 rabbits anesthetized with alpha-chloralose were explored. Monopolar stimulation (30-100 Hz; 0.5 ms pulse duration; 5.0 s train duration; 100-500 microamperemeter) at sites within the central nucleus produced bradycardia and depressor responses. Maximum bradycardia was produced from sites within the anterior, medial component of the nucleus. This response usually began within one second of stimulus onset, reached peak magnitude within two seconds of stimulus onset and in many instances was followed by a slight tachycardia following stimulus termination. The accompanying depressor responses occurred at longer latencies than the bradycardia responses. The responses persisted following artificial ventilation and immobilization by Flaxedil and were attenuated by i.v. injections of atropine methylnitrate. While bradycardia and depressor responses were elicited from sites immediately dorsal, ventral and lateral to the medial central nucleus, component. The results are consistent with previous evidence which suggests a contribution for the central nucleus in the expression of cardiovascular responding during aversive Pavlovian conditioning in the rabbit.

Autonomic and somatomotor effects of amygdala central N. stimulation in awake rabbits

Low level stimulation of the amygdala central nucleus was found to produce bradycardia responses in awake rabbits. Within the central nucleus the most sensitive sites were located within the medial component. The bradycardia response was accompanied by changes in respiration, most commonly an increase in frequency and a decrease in tidal volume, as well as by pupillodilation. Somatomotor responses to central nucleus stimulation were an arrest of ongoing behavior and movements of the mouth and tongue. The pattern of responses observed following stimulation of this nucleus is similar to that observed in response to threatening stimuli and suggests a contribution for this nucleus in the integration of a species-appropriate emotional response in the rabbits.

Amygdaloid D1 dopamine receptor involvement in Pavlovian fear conditioning

The amygdala has long been implicated in conditioned fear. The mesencephalic dopaminergic system provides a rich innervation to the amygdala [J.H. Fallon, P. Ciofi, Distribution of monoamines within the amygdala, in: J.P. Aggleton (Ed.), The Amygdala: Neurobiological Aspects of Emotion, Memory and Mental Dysfunction, Wiley, New York, 1992, pp. 97-114; L.J. Freedman, M.D. Cassell, Distribution of dopaminergic fibers in the central division of the extended amygdala of the rat. Brain Research 633 (1994) 243-252; E. Asan, The catecholaminergic innervation of the rat amygdala. Advances in Anatomy Embryology and Cell Biology 142 (1996) 1-107]. Specific activation of the mesoamygdaloid dopaminergic system has been reported to occur in response to conditioned fear-arousing stimuli [M.L. Coco, C.M. Kuhn, T.D. Ely, C.D. Kilts, Selective activation of mesoamygdaloid dopamine neurons by conditioned stress: attenuation by diazepam. Brain Research 590 (1992) 39-47] suggesting that dopamine release in the amygdala may contribute to the acquisition and/or expression of conditioned fear. Using a 2x2 factorial design, Experiment 1A investigated the effects of bilateral intra-amygdaloid infusions of the selective D1 receptor antagonist, SCH 23390 (2.0 microgram 0.5 microliter-1 side-1), on the acquisition and expression of Pavlovian conditioned fear measured by freezing to acoustic and background contextual stimuli. Infusions of SCH 23390 prior to acquisition training, prior to retention testing or prior to both significantly attenuated conditioned freezing during retention testing. Experiment 1B investigated the dose-dependent effects of pre-training infusions of SCH 23390 (0.5, 1.0 and 2.0 microgram) on conditioned fear. Pre-training infusions of SCH 23390 dose-dependently attenuated conditioned freezing during retention testing. Experiment 2A investigated the effects of bilateral infusions of the selective D1 receptor agonist, SKF 82958 (2.0 microgram 0.5 microliter-1 side-1) on the acquisition and expression of conditioned fear. Infusions of SKF 82958 prior to training facilitated conditioned freezing during retention testing. Experiment 2B investigated the dose-dependent effects of pre-training infusions of SKF 82958 (1.0, 2.0 and 4.0 microgram) on conditioned fear. Pre-training infusions of SKF 82958 dose-dependently facilitated conditioned freezing during retention testing. In conclusion, these results suggest that dopamine transmission within the amygdala contributes to the acquisition and expression of Pavlovian fear conditioning.

An electrophysiological characterization of ventral tegmental area dopaminergic neurons during differential pavlovian fear conditioning in the awake rabbit

Recent research has suggested that the mesencephalic dopaminergic (DA) system is activated by stress. For example, alterations in DA metabolites have been found in the ventral tegmental area (VTA) following footshock and immobilization in the rat [15,37]. Furthermore, this activation appears selective to DA neurons within the VTA since no changes were observed within the substantia nigra [15,16]. While this research suggests that DA neurons in the VTA are activated by aversive events, there has been a paucity of electrophysiological research designed to examine the sensory response characteristics of these DA neurons, and in particular their response to stimuli which predict aversive events. The present study was conducted to investigate the response characteristics of DA neurons within the VTA of the awake rabbit to acoustic stimuli which, via Pavlovian aversive conditioning procedures, came to predict the occurrence of a mild shock to the pinna. 45%, of the neurons meeting pre-established criteria for DA neurons demonstrated either significant excitation or inhibition to conditioned aversive stimuli. These neurons responded differentially to CS+ and CS- presentations. Some of these neurons (65%) demonstrated a greater increase in activity during the CS+ compared to the CS-, some (22%,) demonstrated a greater decrease in activity during the CS+ compared to the CS- and some (13%) demonstrated a greater increase in activity during the CS- compared to the CS+. Further, conditioned heart rate responses in the rabbits occurred during the recording of a majority of these neurons. These overall results suggest that conditioned aversive stimuli can affect the firing of VTA DA neurons and that these neurons comprise a heterogenous population with respect to their response profiles.

The origin and extent of direct amygdala projections to the region of the dorsal motor nucleus of the vagus and the nucleus of the solitary tract

The site of origin of a direct projection from the amygdala to autonomic regulatory nuclei in the dorsomedial medulla has been precisely localized and delimited in the rabbit by means of the retrograde axonal transport of horseradish peroxidase. Labeled cells in the amygdala were found to be confined within regions of the ipsilateral central nucleus. Many labeled cells were also observed to extend beyond the central nucleus into closely related regions of the substantia innominata and bed nucleus of the stria terminalis.

Multiple unit activity recorded from amygdala central nucleus during Pavlovian heart rate conditioning in rabbit.

Using a Pavlovian heart rate conditioning paradigm, a rapid development of short latency increases in the multiple unit activity of the amygdala central nucleus were observed in response to a tone conditioned stimulus. In some cases the increase in multiple unit response showed a parallel development with the conditioned decelerative heart rate response and were significantly correlated with it. These results suggest a direct role for the central nucleus in the expression of conditioned heart rate responding in rabbit.

The effects of intra-amygdaloid infusions of a D2 dopamine receptor antagonist on Pavlovian fear conditioning.

The present study examined the effects of bilateral intra-amygdaloid infusions of the D2 receptor antagonist, eticlopride, on the acquisition and expression of Pavlovian fear conditioning as measured by freezing to acoustic and background contextual stimuli in the rat. Infusions of eticlopride before acquisition or before both acquisition and retention testing significantly attenuated conditioned freezing to tone presentations during the retention test 24 hr later. No effects, however, were observed on freezing that emerged during acquisition. Furthermore, these effects were not attributable to state-dependent learning effects or alterations in baseline activity or shock reactivity. In conclusion, these results suggest that amygdaloid dopamine transmission at D2 receptors contributes to the formation and/or consolidation of fear memories.

Effects of electrical stimulation of the amygdaloid central nucleus on neocortical arousal in the rabbit

This study sought to determine whether electrical stimulation of the amygdaloid central nucleus (ACe) produces cholinergically mediated neocortical arousal manifested in the suppression of frontal cortex delta wave (1-4 Hz) activity. Stimulation in both anesthetized and conscious rabbits produced a suppression of delta activity that was accompanied by bradycardia and blocked by cholinergic antagonists. Stimulation of the adjacent putamen did not produce delta suppression, whereas stimulation of the adjacent ventral globus pallidus produced a suppression of shorter duration than that produced by ACe stimulation. The results suggest that the ACe influences neocortical arousal, which may be mediated by its influence on the activity of cholinergic neurons of the nucleus basalis.