Amygdaloid central nucleus neuronal activity accompanying pavlovian cardiac conditioning: effects of naloxone

Rabbits were treated intravenously with either naloxone-HCl (0.5 mg/kg) or saline vehicle prior to aversive pavlovian conditioning and extinction training; heart rate conditioned responses and concomitant multiple-unit activity in the amygdaloid central nucleus were compared. Multiple-unit activity evoked by the conditioned stimulus increased during conditioning and decreased during extinction in saline-treated rabbits; naloxone treatment attenuated evoked neuronal activity but enhanced bradycardiac conditioned responses. Correlational analysis showed that, in 3 of 8 animals in the saline-treated group, larger increases in multiple-unit activity corresponded to smaller bradycardiac responses. Naloxone treatment did not alter the frequency or magnitude of this relationship, but it did augment the training-induced decrements in evoked neuronal activity at placements that were correlated with bradycardiac response magnitude. These data suggest that conditioned decreases in central nucleus neuronal activity normally may serve to disinhibit vagal mechanisms as conditioned bradycardia develops and that the neural circuits that produce these responses are sensitive to opioid modulation.     

Cryogenic blockade of the central nucleus of the amygdala attenuates aversively conditioned blood pressure and respiratory responses

The central nucleus of the amugdala (ACE) was reversibly blocked during extinction of an aversively conditioned cardiorespiratory response in unanesthetized, freely moving cats. Cryoprobes were positioned bilaterally in the ACE of 4 cats and in the nucleus entope-duncularis of 1 cat. Blood pressure typically showed biphasic changes in response to the conditioned stimulus (CS) during non-cooling trials. Blood pressure initially dropped and then rose. Heart rate consistently dropped, and respiratory rate increased in response to the CS. ACE cooling did not alter the pre-CS baseline blood pressure, heart rate or respiratory timing, but changed the cardiorespiratory response to the CS. During ACE cooling, blood pressure and respiratory responses were greatly attenuated or abolished. No significant effect on the heart rate response was observed during ACE cooling. Cooling of a nearby structure, the nucleus entopeduncularis, did not affect blood pressure, heart rate or respiratory responses to the CS. These results support the hypothesis that the ACE plays a role in both cardiovascular and respiratory regulation during conditioned aversive responses. The study also suggests that, in cats, the predominant influence of the ACE on cardiovascular control is on blood pressure rather than on heart rate regulation.     

Cardiovascular responses elicited by stimulation of neurons in the central amygdaloid nucleus in awake but not anesthetized rats resemble conditioned emotional responses

Cardiovascular responses elicited by electrical stimulation of the central amygdaloid nucleus were examined in awake and anesthetized rats. Stimulation through chronically implanted electrodes evoked increases in arterial pressure and heart rate in awake, freely behaving rats. The responses, which were dependent upon the frequency and the intensity of the stimulus, were not consistently related to the presence of evoked amygdaloid afterdischarges or to evoked behavioral reactions. Following induction of anesthesia, stimuli delivered to the same rats through the same fixed electrodes produced decreases in blood pressure and heart rate. Microinjection ofl-glutamate into the amygdala of freely behaving rats also elicited increases in arterial pressure and heart rate, indicating that the cardiovascular changes evoked by electrical stimuli are due to excitation of local neurons rather than fibers of passage. The timing and pattern of the response elicited by electrical stimulation of the amygdala in the awake but not anesthetized rat closely corresponds with that evoked by an acoustic conditioned emotional stimulus.     

The discharge characteristics of vagal cardiac neurons during classically conditioned heart rate change

Classically conditioned heart rate change in the pigeon has been developed as a vertebrate model system for cellular analysis of associative learning. One aspect of this development involved characterizing in detail the "final common path" for the conditioned response, the "cardiac motoneurons." With this as a foundation it has been possible to investigate the discharge properties of these motoneurons during acquisition of the conditioned heart rate response, and in this paper we report such data for the vagal (parasympathetic) component of the conditioned response. The activity of single neurons was recorded in the intermediate zone of the dorsal motor nucleus of the vagus of the pigeon. Antidromic activation confirmed that these cells gave rise to efferent axons in the vagus nerve and allowed their classification on the basis of conduction velocity into three groups: A, B1 (cardiac), and B2 units. B1 and B2 units had low levels of irregularly maintained activity, whereas A units discharged more regularly at higher frequency. B1 units had a weak cardiac rhythm, no respiratory rhythm, and showed decreased discharge in response to visual stimulation or foot-shock. These stimulus-evoked decreases in the activity of cardiac cells were accompanied by increases in heart rate. B2 units were unresponsive to both visual stimulation and foot-shock, whereas A units were unresponsive to visual stimulation but possibly responsive to foot-shock. The activity of vagal cardiac neurons was recorded during or after classical conditioning (paired lights and foot-shocks) or sensitization (unpaired lights and foot-shocks) to characterize discharge changes associated with conditioned cardioacceleration.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of lidocaine injection in the interpositus nucleus and red nucleus on conditioned behavioral and neuronal responses

The role of the cerebellum and the red nucleus in the conditioned eyeblink response was assessed, using a combination of reversible lesions and multiple-unit extracellular recording in the awake, behaving rabbit. Lesion, recording, and stimulation experiments have indicated that both of these structures are involved in the performance of learned skeletal muscle responses. The present study sought to distinguish the relative contributions of the interpositus nucleus and the red nucleus to the expression of the learned response by recording behavior-related multiple unit activity in one structure while reversibly inactivating the other via injections of local anesthetic. Results indicate that inactivating either the interpositus or the red nucleus temporarily abolishes the learned eyeblink response. Injection of lidocaine into the interpositus also abolishes the neuronal unit model of the conditioned response in the red nucleus, while injection into the red nucleus does not affect the model in the interpositus. These results are consistent with the hypothesis that the red nucleus acts as a relay for motor commands from the cerebellum, and that the plasticity that generates conditioned responses occurs in the cerebellum or an afferent structure.     

Neuronal activity in the midline thalamic nuclei during pavlovian heart rate conditioning

Multiple unit activity (MUA) was recorded from chronically implanted electrodes in the midline nuclei of the thalamus in rabbits receiving either Pavlovian heart rate (HR) conditioning, or explicitly unpaired presentations of the conditioned stimulus (CS) and unconditioned stimulus. Animals receiving paired training demonstrated increases in CS-evoked MUA compared to animals receiving unpaired training; however, these increases were small and only weakly correlated with the HR conditioned response, suggesting that the midline nuclei are only minimally, if at all, involved in associative learning.     

Inhibition of the amygdala central nucleus by stimulation of cerebellar output in rats: a putative mechanism for extinction of the conditioned fear response

The amygdala and the cerebellum serve two distinctively different functions. The amygdala plays a role in the expression of emotional information, whereas the cerebellum is involved in the timing of discrete motor responses. Interaction between these two systems is the basis of the two‐stage theory of learning, according to which an encounter with a challenging event triggers fast classical conditioning of fear‐conditioned responses in the amygdala and slow conditioning of motor‐conditioned responses in the cerebellum. A third stage was hypothesised when an apparent interaction between amygdala and cerebellar associative plasticity was observed: an adaptive rate of cerebellum‐dependent motor‐conditioned responses was associated with a decrease in amygdala‐dependent fear‐conditioned responses, and was interpreted as extinction of amygdala‐related fear‐conditioned responses by the cerebellar output. To explore this hypothesis, we mimicked some components of classical eyeblink conditioning in anesthetised rats by applying an aversive periorbital pulse as an unconditioned stimulus and a train of pulses to the cerebellar output nuclei as a cerebellar neuronal‐conditioned response. The central amygdala multiple unit response to the periorbital pulse was measured with or without a preceding train to the cerebellar output nuclei. The results showed that activation of the cerebellar output nuclei prior to periorbital stimulation produced diverse patterns of inhibition of the amygdala response to the periorbital aversive stimulus, depending upon the nucleus stimulated, the laterality of the nucleus stimulated, and the stimulus interval used. These results provide a putative extinction mechanism of learned fear behavior, and could have implications for the treatment of pathologies involving abnormal fear responses by using motor training as therapy.     

Nucleus basalis involvement in conditioned neuronal responses in the rat frontal cortex

Rat frontal cortex neurons exhibit alterations in firing in response to a 2 sec tone cue followed by rewarding medial forebrain bundle (MFB) stimulation. Nucleus basalis neurons supply up to 75% of the cortical cholinergic innervation. The nucleus basalis and ACh have been implicated as playing a role in cognitive function. Three experiments were designed to test the hypothesis that the nucleus basalis cholinergic system is involved in the generation of conditioned neuronal responses in the rat frontal cortex. Local microinjection of the cholinergic antagonist, atropine, into the frontal cortex suppressed the conditioned responses of 22 of 25 cortical single units. Unilateral kainic acid lesioning of the nucleus basalis resulted in a significant decrease in the proportion of units exhibiting conditioned responses in the cortex ipsilateral to the lesion (25%) compared to the proportion of responding units from the cortex of untreated animals (70%). When the firing rates of units encountered in the region of the nucleus basalis were monitored during presentation of the cue-MFB paradigm, 28 of 38 unit recordings exhibited significant increases or decreases in firing rate. Therefore, the results of the experiments indicate that the nucleus basalis cholinergic neurons are involved in the generation of conditioned neuronal responses in the rat frontal cortex.     

Cardiovascular effects of α-adrenergic agents in the dorsal raphe nucleus of the rat

Local injection of norepinephrine (NE) or phenylephrine into the lateral aspect of the dorsal raphe nucleus results in an increase in blood pressure and heart rate in the urethane-anesthetized rat. The increases in blood pressure and heart rate are blocked by prior injection of phentolamine into the dorsal raphe nucleus and are significantly reduced by i.v. mecamylamine. Selective lesion of serotonergic neurons in the dorsal raphe nucleus by 5,7-dihydroxytryptamine significantly reduces the pressor response to phenylephrine, but does not affect the increase in heart rate in response to phenylephrine. These data are consistent with a central α-adrenergic mechanism in the dorsal raphe that elevates blood pressure at least partly by an action on serotonergic neurons and elevates heart rate by a mechanism involving non-serotonergic neurons in the dorsal raphe area.     

Accessory abducens nucleus and conditioned eye retraction/nictitating membrane extension in rabbit

The role of accessory abducens nucleus neurons in the conditioned eye retraction/nictitating membrane extension response was defined in the rabbit. Horseradish peroxidase injections into the retractor bulbi muscle showed that accessory abducens nucleus is the principal location of its motor-neurons. Single and multiple unit recording in accessory abducens indicated that these motor neurons show a marked responsiveness to corneal and periorbital stimulation and fire in close correlation with conditioned, unconditioned, or spontaneous eye retraction/nictitating membrane extension. Complete lesions of accessory abducens showed, at most, a partial reduction of the conditioned and unconditioned eye retraction response. Section of the extraocular muscles, other than retractor bulbi, also caused a partial reduction of the eye retraction response. Accessory abducens lesions, combined with extraocular muscle section, were necessary to dramatically reduce the eye retraction response permanently. These experiments demonstrated that accessory abducens is a primary controller of eye retraction through its axons to retractor bulbi. The other extraocular muscles act in concert with retractor bulbi to elicit conditioned and unconditioned eye retractions.     

Ibotenic acid lesions in the amygdaloid central nucleus but not in the lateral subthalamic area prevent the acquisition of differential Pavlovian conditioning of bradycardia in rabbits

The present study examined the effect of ibotenic acid lesions in the amygdaloid central nucleus (ACe) or in the lateral zona incerta of the subthalamus (LZI) on the acquisition of differential Pavlovian conditioning of bradycardia in rabbits. Previous work has shown that bilateral electrolytic lesions in either ACe or LZI abolished the retention of conditioned heart rate (HR) responses. In order to determine whether these findings were due to destruction of cells intrinsic to ACe or LZI, ibotenic acid lesions were placed bilaterally in either structure or in control sites. Following recovery, animals were subjected to differential Pavlovian conditioning in which one tone (CS+) was paired with periorbital shock and a second tone (CS−) was presented alone. It was found that destruction of cell bodies in ACe, but not LZI, prevented the acquisition of the differential bradycardiac conditioned response. In addition, ACe lesions did not interfere with baseline HR, the HR orienting response, the HR unconditioned response to shock, or the concomitantly conditioned corneoretinal potential. The results of this study suggest that destruction of cells intrinsic to ACe selectively prevents the acquisition of differentially conditioned HR, and perhaps other conditioned responses related to conditioned arousal, but does not affect unlearned HR responses or specific somatomotor conditioned responses.     

Parasagittal thalamic knife cuts retard pavlovian eyeblink conditioning and abolish the tachycardiac component of the heart rate conditioned response

Rabbits received parasagittal knife cuts lateral to the mediodorsal nucleus of the thalamus (MD), severing afferents and efferents to and from the prefrontal cortex. These animals were compared to sham animals in a Pavlovian eyeblink and heart rate conditioning experiment in which a tone was the conditioned stimulus (CS) and paraorbital electrical shock was the unconditioned stimulus (US). Knife cuts retarded acquisition of the eyeblink conditioned response (CR), and abolished the late-occurring tachycardiac component of the heart rate CR. These data are compatible with previous experiments which suggest that MD participates in the sympathetic control associated with somatomotor learning.     

Different projections of the central amygdaloid nucleus mediate autonomic and behavioral correlates of conditioned fear

The purpose of the present study was to determine whether lesions of areas projected to by the central amygdaloid nucleus (ACE) would disrupt the classical conditioning of autonomic and/or behavioral emotional responses. The areas studied included 3 projection targets of the ACE: the lateral hypothalamic area (LH), midbrain central gray (CG) region, and bed nucleus of the stria terminalis (BNST). Lesions were made either electrolytically or by microinjection of ibotenic acid, which destroys local neurons without interrupting fibers of passage. Two weeks later, the animals were classically conditioned by pairing an acoustic stimulus with footshock. The next day, conditioned changes in autonomic activity (increases in arterial pressure) and emotional behavior ("freezing," or the arrest of somatomotor activity) evoked by the acoustic conditioned stimulus (CS) were measured during extinction trials. Electrolytic and ibotenic acid lesions of the LH interfered with the conditioned arterial pressure response, but did not affect conditioned freezing. Electrolytic lesions of the rostral CG disrupted conditioned freezing but not conditioned changes in arterial pressure. Ibotenic acid injected into the rostral CG reduced neither the arterial pressure nor the freezing response. Injection of ibotenic acid in the caudal CG, like electrolytic lesions of the rostral CG, disrupted the freezing, but not the arterial pressure response. Injection of ibotenic acid into the BNST had no effect on either response. These data demonstrate that neurons in the LH are involved in the autonomic, but not the behavioral, conditioned response pathway, whereas neurons in the caudal CG are involved in the behavioral, but not the autonomic, pathway. Different efferent projections of the central amygdala thus appear to mediate the behavioral and autonomic concomitants of conditioned fear.     

Vasopressinergic modulation of stress responses in the central amygdala of the Roman high-avoidance and low-avoidance rat.

The central nucleus of the amygdala (CEA) is selectively involved in the passive component of the behavioral (immobility) and the accompanying parasympathetic response during conditioned, stressful environmental challenges. Vasopressinergic mechanisms in the brain seem to play a role in these stress responses. The effects of the neuropeptides arginine-8-vasopressin (AVP) and oxytocin (OXT) on modulating CEA activity during conditioned stress of inescapable footshock were studied in male Roman high-avoidance (RHA/Verh) and low-avoidance (RLA/Verh) rats, psychogenetically selected on the basis of shuttle-box acquisition behavior. In RLA/Verh rats, the cardiac and behavioral responses to the conditioned emotional stressor were bradycardia and immobility, suggesting an important role for the CEA in these rats. The RHA/Verh rats, however, failed to show any change in heart rate or immobility in response to a conditioned stress situation. The low dose of AVP (20 pg) in the CEA of conscious RLA/Verh rats caused an enhancement of the stress-induced bradycardiac and immobility response. However, the high dose of AVP (2 ng) and OXT (200 pg) attenuated the bradycardiac and immobility responses in the RLA/Verh rats. Infusion of AVP and OXT in the RHA/Verh rats failed to induce any change in heart rate or immobility. Binding studies revealed that the AVP receptor selectively binds AVP with high affinity. In contrast, the OXT receptor recognizes both AVP and OXT with a similar (but lower) affinity. This suggests that the behavioral and autonomic responses of the high dose of AVP may be caused by OXT receptor stimulation.(ABSTRACT TRUNCATED AT 250 WORDS)     

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.     

Multiple response measures during classical conditioning

Animal research assessing multiple responses during Pavlovian conditioning has revealed a dichotomy between the central nervous system (CNS) substrates for somatomotor and visceral CRs. These findings have implications for the study of clinical/applied problems in human subjects, since differences in the acquisition functions for these response systems may suggest which CNS structures are involved in various neuropsychiatric disorders. The present paper describes methods and procedures utilized to assess the somatomotor conditioned eyeblink (EB) response and accompanying visceral changes in human subjects. Methods are described for assessing concomitant EB conditioned and unconditioned responses and the accompanying heart rate, skin conductance, and respiratory changes during Pavlovian conditioning in human subjects. It is stressed that utilization of concomitant conditioning of these different response systems may lead to inferences regarding the central nervous system structures involved in a variety of different kinds of clinical problems.     

Differential effect of lesioning of the central amygdala on the bradycardiac and behavioral response of the rat in relation to conditioned social and solitary stress

The central nucleus of the amygdala (CEA) is considered to be involved in stress-dependent regulation of autonomic functions. In the present study the effects of a bilateral electrolytic lesion of the CEA on the cardiac response and on the duration of immobility behavior in two conditioned stress situations have been analysed in Wistar rats. Fear of a previously received electric footshock elicited in almost all animals a profound early bradycardia and a marked immobility behavior in comparison to non-stressed freely moving controls. Lesioning of the CEA completely abolished the bradycardiac response. Immobility behavior was slightly diminished. The conditioned fear of a dominant rat following social defeat was accompanied by a bradycardiac response in about 50% of the rats. This bradycardiac response was not accompanied by significant immobility behavior. The rest of the defeated rats--i.e. cardiac 'non-responders'--displayed an early increase in immobility behavior. The CEA lesion abolished again the bradycardiac response, but the immobility behavior remained unchanged. In the cardiac 'non-responders' CEA lesion failed to affect immobility behavior, and the heart rate also remained unchanged. These results suggest that the CEA is involved in the organization of conditioned parasympathetic responses independent of the nature of stress. In contrast, the role of this nucleus in the organization of the immobility behavior seems to depend on the nature of stress and other, probably individual characteristics of the rats' behavior.     

Modification of neuronal discharge along the ascending tectofugal pathway during visual conditioning

Visually conditioned heart-rate change in the pigeon has been developed as vertebrate model system for analysis of associative learning. The visual pathways transmitting the conditioned stimulus information were identified, and neurophysiological analyses during conditioning were then undertaken to determine if these pathways behave merely as input lines or undergo training-induced modification. After finding that the retinal output is invariant with training, we investigated the central visual pathways, beginning with the tectofugal pathway. During conditioning single neurons in the nucleus rotundus and ectostriatum, the thalamic and telencephalic relays of the tectofugal pathway, showed enhancement of their phasic light-evoked responses. In contrast, the initial phasic responses attenuated during non-associative training. The rate at which these discharge modifications developed paralleled the development of the behavioral response. Thus, the tectofugal pathway shows plasticity during conditioning and does not behave merely as an input channel for the conditioned stimulus.     

Involvement of the avian hypothalamus in defensively conditioned heart rate change.

Hypothalamic involvement in visually conditioned heart rate change (established by pairing light and foot-shock) was studied in 128 pigeons by evaluating conditioning performance following lesions of various hypothalamic areas. Extensive destruction of the posterior hypothalamus severely impaired development of conditioned heart rate change. Anterior hypothalamic lesions also produced serious deficits, though not as severe as following posterior hypothalamic damage. Partial posterior hypothalamic, unilateral and tuberal lesions produced only minor to moderate deficits. The critical locus for profound impairment of conditioned response development appeared to be the medial hypothalamus, and it is suggested that more specifically it is the terminal field of the archistriatal projection upon the medial hypothalamus. It is concluded that the medial hypothalamus is essential for the development of defensively conditioned heart rate change, and based on previous findings it is suggested that the critical descending pathway for expression of this conditioned response involves the archistriatal projection upon the medial hypothalamus and subsequently a polysynaptic pathway through the ventral brainstem.     

Hippocampal cellular plasticity during extinction of classically conditioned nictitating membrane behavior

Hippocampal unit responses were recorded during extinction of classically conditioned nictitating membrane (NM) behavior in the rabbit. Prior studies have shown that during the acquisition phase of nictitating membrane conditioning, the frequency of hippocampal cell firing increases at a faster rate (across trials) than learned behavior. Results reported here show that, during the early phases of extinction, conditioned hippocampal unit responses decrement at a faster rate than learned NM behavior. Furthermore, only certain components of the conditioned hippocampal unit response display robust spontaneous recovery across successive days of extinction training. In all, results show that changes in the activity of hippocampal neurons predict changes in learned behavior over trials of conditioning and extinction, and demonstrate that hippocampal cellular activity is particularly sensitive to stimulus configurations or environmental contingencies that produce changes in behavior.