Retrieving fear memories, as time goes by .

Research in fear conditioning has provided a comprehensive picture of the neuronal circuit underlying the formation of fear memories. In contrast, our understanding of the retrieval of fear memories is much more limited. This disparity may stem from the fact that fear memories are not rigid, but reorganize over time. To bring some clarity and raise awareness about the time-dependent dynamics of retrieval circuits, we review current evidence on the neuronal circuitry participating in fear memory retrieval at both early and late time points following auditory fear conditioning. We focus on the temporal recruitment of the paraventricular nucleus of the thalamus (PVT) for the retrieval and maintenance of fear memories. Finally, we speculate as to why retrieval circuits change with time, and consider the functional strategy of recruiting structures not previously considered as part of the retrieval circuit.

Fear conditioning enhances different temporal components of tone-evoked spike trains in auditory cortex and lateral amygdala.

Single neurons were recorded in freely behaving rats during fear conditioning from areas of auditory cortex that project to the lateral nucleus of the amygdala (LA). The latency and rate of conditioning and extinction were analyzed, and the results were compared to previous recordings from LA itself. Auditory cortex neurons took more trials to learn, and they responded more slowly than LA neurons within trials. Short-latency plasticity in LA, therefore, reflects inputs from the auditory thalamus rather than the auditory cortex. Unlike LA cells, some auditory cortex cells showed late conditioned responses that seemed to anticipate the unconditioned stimulus, while others showed extinction-resistant memory storage. Thus, rapid conditioning of fear responses to potentially dangerous stimuli depends on plasticity in the amygdala, while cortical areas may be particularly involved in higher cognitive (mnemonic and attentional) processing of fear experiences.

Fear conditioning enhances short-latency auditory responses of lateral amygdala neurons: parallel recordings in the freely behaving rat.

The lateral nucleus of the amygdala (LA) is the first site in the amygdala where the plasticity underlying fear conditioning could occur. We simultaneously recorded from multiple LA neurons in freely moving rats during fear conditioning trials in which tones were paired with foot shocks. Conditioning significantly increased the magnitude of tone-elicited responses (often within the first several trials), converted unresponsive cells into tone-responsive ones, and altered functional couplings between LA neurons. The effects of conditioning were greatest on the shortest latency (less than 15 ms) components of the tone-elicited responses, consistent with the hypothesis that direct projections from the auditory thalamus to LA are an important link in the circuitry through which rapid behavioral responses are controlled in the presence of conditioned fear stimuli.

Consolidation of extinction learning involves transfer from NMDA-independent to NMDA-dependent memory.

Extinction of conditioned fear to a tone paired with foot shock is thought to involve the formation of new memory. In support of this, previous studies have shown that extinction of conditioned fear depends on NMDA receptor-mediated plasticity. To further investigate the role of NMDA receptors in extinction, we examined the effects of the NMDA antagonist d(-)-3-(2-carboxypiperazine-4-yl)-propyl-1-phosphonic acid (CPP) on the extinction of conditioned freezing and suppression of bar pressing (conditioned emotional response). Rats extinguished normally during a 90 min session in the presence of systemic CPP (10 mg/kg), but were unable to recall extinction learning 24 hr later. This suggests that an NMDA-independent form of plasticity supports short-term extinction memory, but NMDA receptors are required for consolidation processes leading to long-term extinction memory. Surprisingly, extinction learned in the presence of CPP was recalled normally when tested 48 hr after training, suggesting a delayed consolidation process that was able to improve memory in the absence of further training. Delayed consolidation involves NMDA receptors because CPP injected on the rest day between training and test prevented 48 hr recall of extinction learned under CPP. Control experiments showed that the effect of CPP on memory consolidation was not caused by state-dependent learning or reduced expression of freezing under CPP. These findings demonstrate that NMDA receptor activation is critical for consolidation of extinction learning and that this process can be initiated after training has taken place. We suggest that consolidation of extinction involves off-line relearning that reinforces extinction memory through NMDA-mediated plasticity, perhaps in prefrontal-amygdala circuits.

Amphetamine sensitization is accompanied by an increase in prelimbic cortex activity.

Drug addiction is associated with dysfunction in the medial prefrontal cortex (mPFC). However, the modifications of neuronal activity in mPFC underlying the reinforcing properties of addictive drugs are still unclear. Here we carried out single-unit recording experiments to study the neuronal activity in the prelimbic (PL) cortex of anesthetized rats, after expression of locomotor sensitization to amphetamine. In control rats, an acute injection of amphetamine induced mainly an inhibitory effect on firing rate (FR) and this response was negatively correlated with the basal FR. Sensitized rats showed a higher proportion of excited neurons and the response to amphetamine was independent of basal FR. Moreover, in control rats, acute amphetamine decreased burst rate, whereas in sensitized rats acute amphetamine increased burst rate. These findings indicate that amphetamine sensitization renders mPFC neurons hyperexcitable. Taken together, these data support the hypothesis that early withdrawal is associated with an increase in the activity of the mPFC, which could strengthen the PL-Nucleus Accumbens connection, thus facilitating amphetamine-induced locomotor sensitization.

Central somatosensory conduction time in severely growth-stunted children.

To examine the effects of chronic malnutrition on central nervous system function, we used the somatosensory evoked potential to measure the central conduction time of 20 children aged 7-8 y with heights below the third percentile for their age and 20 control children in Honduras. The two groups differed significantly in socioeconomic status, achievement in Bender's neurointegrative test, and hematocrit, but not in birth weight. After median nerve stimulation, the mean central conduction time (interpeak latency between N13 and N20) for the growth-stunted group (6.19 +/- 0.52 ms) did not differ significantly from that of the control subjects (6.30 +/- 0.58 ms), suggesting appropriate myelination and fiber diameter. Somatosensory tracts may escape damage resulting from postnatal dietary deficiencies because myelination in these tracts is almost complete at birth.

Mechanism of action of a new prostaglandin antihypertensive, viprostol [CL 115 347; (dl)-15-deoxy-16-hydroxy-16(alpha/beta)-vinyl-prostaglandin E2 methyl ester]: (II). Effects on the adrenergic nervous system.

Viprostol [(dl)-15-deoxy-16-hydroxy-16(alpha/beta)-vinyl-prostaglandin E2 methyl ester; CL 115 347] is a new orally and transdermally active antihypertensive agent that exerts its major antihypertensive action by vasodilation. The present studies were conducted to examine its effects on the adrenergic nervous system. In cats, viprostol did not inhibit renal sympathetic nerve discharge (RSND) monitored at the postganglionic region, indicating that nerve transmission or conduction was not blocked at the ganglion or the pre- or postganglionic fibres. In cat nictitating membrane preparations in situ, viprostol partially blocked the membrane contractile response to exogenous epinephrine and norepinephrine, as well as to electrical stimulation of pre- and postganglionic fibres. In spontaneously hypertensive rats (SHR), viprostol partially blocked the vasopressor response of exogenous norepinephrine and epinephrine specifically without influencing that of angiotensin II. All these suggest that viprostol produced weak alpha-adrenoceptor blockade. Viprostol did not antagonize the tachycardia induced by stimulation of the discrete segments at C7-T1 (cardio-accelerator) of the spinal cord in pithed SHR, suggesting that viprostol did not activate the presynaptic alpha-adrenoceptors. Viprostol significantly inhibited the increase in blood pressure induced by electrical stimulation of the spinal cord at T7-T9 in pithed SHR, probably due to postsynaptic alpha-adrenoceptor blockade. In conclusion, viprostol produced weak, but statistically significant alpha-adrenoceptor blockade which may contribute partially to its antihypertensive action.

Evidence for the lack of interaction between (+/-)-1-O-octadecyl-2-acetylglyceryl-3-phosphorylcholine and alpha-adrenoceptors in vivo and in vitro.

The interactions of (+/-)-1-O-octadecyl-2-acetylglyceryl-3-phosphorylcholine (octadecyl-AGPC) with alpha-adrenoceptors were studied in rat mesenteric artery, cat nictitating membrane and on the blood pressure of the cat and spontaneously hypertensive (SH) rat. Using a direct radioligand alpha-adrenoceptor binding assay in particulate fractions of rat mesenteric arteries, octadecyl-AGPC was found to be 5 X 10(7) and 75 times less potent than prazosin and noradrenaline (NA), respectively, in displacing (2,6-dimethoxyphenoxyethyl)-aminomethyl-1,4-benzodioxane ([3H]-WB 4101--a selective probe for the identification of alpha-adrenoceptors). In the cat, intravenous infusions of octadecyl-AGPC, which produce a hypotensive response, did not attenuate nictitating membrane contractions in vivo in response to intravenous injections of NA, adrenaline (Ad) or to electrical stimulation of the postganglionic fibres of the superior cervical ganglion. In these experiments, the pressor responses to NA or Ad were not affected by octadecyl-AGPC. Phentolamine, on the other hand, attenuated nictitating membrane contractions and blood pressure responses to Ad or NA. In the SH rat, octadecyl-AGPC decreased mean arterial blood pressure (MABP). After an intravenous dose of phentolamine which lowered MABP, the depressor response to octadecyl-AGPC was reduced. When MABP in the phentolamine-treated SH rat was restored to its initial level with an infusion of angiotensin II (AII), the depressor response to octadecyl-AGPC was restored to its original magnitude. The effectiveness of alpha-adrenoceptor blockade under these experimental conditions was monitored with intravenous NA and Ad. Thus, based on radioligand binding studies and pharmacological studies, it is concluded that octadecyl-AGPC lacks the ability to interact with alpha-adrenoceptors.

Electrical stimulation of medial prefrontal cortex reduces conditioned fear in a temporally specific manner.

The authors recently showed that extinction of auditory fear conditioning leads to potentiation of tone-evoked activity of neurons in the infralimbic (IL) subregion of the medial prefrontal cortex, suggesting that IL inhibits fear after extinction (M. R. Milad, & G. J. Quirk, 2002). In support of this finding, pairing conditioned tones with brief (300-ms) electrical stimulation of IL reduces conditioned freezing. The present study showed that IL stimulation inhibits freezing if given 0.1 s after tone onset (the latency of tone-evoked responses) but has no effect if given either 1 s before or 1 s after tone onset. This suggests that IL gates the response of downstream structures such as the amygdala to fear stimuli.

Separable roles of hippocampal granule cells in forgetting and pyramidal cells in remembering spatial information.

To investigate the roles individual hippocampal cell groups play in processing of spatial information for memory, we administered low-intensity electrical stimulation to the granule cells, CA3 and CA1 pyramidal cells of the dorsal hippocampus at selected times before and after acquisition of the solution to a radial maze win-stay task. Stimulation of any of the 3 cells populations yielded a variable duration anterograde disruption of memory performance, while stimulation of dentate gyrus granule cells alone produced a declarative memory-specific retrograde amnesia. The amnestic effect of granule cell stimulation was not associated with after discharges in the hippocampus and was prevented by systemic administration of the opiate antagonist naloxone. Our results support the view that this electrical stimulus does not disrupt, but rather, activates the normal function of the granule cell system, resulting in erasure of information held in declarative memory. In contrast, similar activation of the pyramidal cell system does not yield retrograde amnesia, suggesting a normal role for these cells in promoting memory for spatial information.

Corticospinal responses to electrical stimulation of motor cortex in the rat.

Direct and indirect corticospinal responses to electrical stimulation of motor cortex were identified in urethane-anesthetized rats. 'Killed-end' recordings were taken from the corticospinal tract at the level of the cervical cord (C1-C2) and from the medullary pyramid. The identities of direct (D) and indirect (I) corticospinal responses were confirmed by: (1) removing motor cortex to eliminate I activity, and (2) pharmacologically increasing neocortical excitability, prior to any lesions, to increase I activity. Our data indicate that the conduction velocity of the fastest corticospinal fibers is approximately 18 m/s. Our identification of the components of the corticospinal response will permit the interpretation of the more complicated surface or 'non-killed-end' depth recordings which have shown particular utility in evaluating spinal cord damage.

Early malnutrition followed by nutritional restoration lowers the conduction velocity and excitability of the corticospinal tract.

The physiological sequelae of undernutrition were investigated in rats that were undernourished from day 1-21 and subsequently free-fed to 75 days of age. Population responses were recorded in the corticospinal tract following surface stimulation of the motor cortex, which activates corticospinal cells directly, and also indirectly via cortical synapses. The conduction velocity of the fastest corticospinal fibers in 15 malnourished rats was 16.9 m/s, significantly slower (P < 0.001) than the 20.0 m/s observed in 26 controls. In addition, the excitability of corticospinal neurons to direct stimulation was reduced as much as 67% in malnourished rats, while no effect on synaptic activation was observed. Our findings suggest that early malnutrition reduces the number of large fibers in the adult corticospinal tract. These results are discussed with respect to known morphological and behavioral effects of malnutrition in rats and their relevance to humans.

Physiological basis of motor effects of a transient stimulus to cerebral cortex.

This contribution includes a selective review of previously published material, findings from some new experiments, and discussion of some relationships between animal and recent human data. The major questions are: What descends from the cerebral cortex after a brief surface stimulus? What explains the various components of the corticofugal discharge? What are the motor consequences of the corticofugal discharge, and what are the effects of lesions on both? The focus is on the corticospinal system, which through its monosynaptic connection with alpha motoneurons of distal muscles accounts for the short latency movements after a transient cortical stimulus. The pyramidal and lateral corticospinal tract response in monkey or cat to a surface stimulus applied to area 4 is a direct (D) wave conducted in fast axons followed by several indirect (I) waves with a period of greater than 1 ms. Although computer summing reveals, at increasing amplitudes, D and I waves in recordings from nuchal skin, vertebra, and surface of the spinal cord, "killed end" recording is essential to reveal the true extent of I relative to D waves. The D wave might result from excitation of: the initial segment (IS), i.e., the classical spike trigger zone; the first or deeper nodes in white matter; or arborizations of the axon collaterals in gray matter. Under different circumstances, each of these modes of excitation can be effective. Thus, with threshold stimulation through separated bipolar electrodes, intracellular recording from pyramidal tract (PT) and uninvaded motor cortical neurons shows that D activation usually occurs when the membrane potential immediately before the stimulus is relatively depolarized, implying excitation of the IS region, i.e., close to the site of synaptic transfer. A monopolar surface (+) stimulus at the appropriate focus usually generates a D wave at weaker intensity than does a surface (-) stimulus. However, if a little above threshold, stimuli of either polarity generate both D and I waves, but the ratio of D:I amplitude is usually greater with surface (+) stimulation. A theoretical estimate of the depth of excitation by a surface (+) stimulus was consistent with threshold excitation occurring at the first node. Slow PT neurons are excited by surface stimulation, but trivially contribute to population PT or corticospinal recordings. Intracellular recording from PT neurons identifies a monosynaptic excitatory postsynaptic potential as the cause of the first I wave, the period between successive I waves reflecting single delays for synaptic discharge.(ABSTRACT TRUNCATED AT 400 WORDS)

Stress disorders of families of the disappeared: a controlled study in Honduras.

The effect of forced disappearance on the physical and psychological health of family members was assessed by interviews carried out in Honduras. Families of the disappeared were compared with two control groups: (1) families who lost a member due to accident or illness; and (2) families where no one had died within the past 10 years. Constellations of stress-related symptoms commonly seen in post-traumatic stress disorder and other anxiety disorders were approx. 2 times more prevalent in families of the disappeared as compared to the other two groups, indicating that families of the disappeared suffer over and above that due to normal grieving. It is suggested that the atmosphere of fear and isolation experienced by families of the disappeared is a causative factor in the prolongation of stress-related disorders years after the traumatic event.

Starting a neuroscience research laboratory in a developing country: a Fulbright experience in Honduras.

The first laboratory of neurophysiology was installed in the medical school of the University of Honduras during the 1992-1993 academic year. The goal of the project was to improve the teaching of physiology in the medical curriculum and to establish a neuroscience research laboratory able to address Honduran needs. In addition to a computer learning facility and wet labs in neurophysiology for medical students, an independent research program that focused on social problems in the country (for example, the effects of malnutrition on the developing central nervous system) was developed, paving the way for the first graduate program in physiology in Honduras. Funded by a Fulbright Lectureship Grant, the shoe-string budget was augmented by donations of equipment by colleagues. This first-hand account describes the planning and implementation of the project, covering both expected and unexpected problems and successes. An update on the progress of the lab after two years of independent operation is also described.

The firing of hippocampal place cells in the dark depends on the rat's recent experience.

Hippocampal "place cells" fire when a freely moving rat is in a given location. The firing of these cells is controlled by visual and nonvisual environmental cues. The effects of darkness on the firing of place cells was studied using the task of Muller et al. (1987), in which rats were trained to chase randomly scattered food pellets in a cylindrical drum with a white cue-card attached to the wall. The position of the rats was tracked via an infrared LED on the headstage with a video system linked to computer. Two experimental protocols were used: in the first, lights were turned off after the rat had already been placed in the chamber; in the second, the rat was placed in the darkened chamber. The dark segments produced by these 2 methods were identical with respect to light and other cues but differed with respect to the rat's experience. The firing patterns of 24 of 28 cells were unaffected by darkness when it was preceded by a light period. In contrast, the firing patterns of 14 of 22 cells changed dramatically when the rats were put into the darkened chamber. Furthermore, the majority of cells that changed their firing pattern in initial darkness maintained that change when the lights were turned on. These results show that place cells can fire differently in identical cue situations and that the best predictor of firing pattern is a combination of current cues and the rat's recent experience. The results are discussed in terms of mnemonic properties of hippocampal cells and "remapping" of place cell representations.

Differential effects of amygdala lesions on early and late plastic components of auditory cortex spike trains during fear conditioning.

In auditory fear conditioning, pairing of a neutral acoustic conditioned stimulus (CS) with an aversive unconditioned stimulus (US) results in an enhancement of neural responses to the CS in the amygdala and auditory cortex. It is not clear, however, whether cortical plasticity governs neural changes in the amygdala or vice versa, or whether learning in these two structures is determined by independent processes. We examined this issue by recording single-cell activity in the auditory cortex (areas Te1, Te1v, and Te3) of freely behaving, amygdalectomized rats using a movable bundle of microwires. Amygdala damage did not affect short-latency (0-50 msec) tone responses, nor did it interfere with conditioning-induced increases of these onset responses. In contrast, lesions of the amygdala interfered with the development of late (500-1500 msec) conditioned tone responses that were not present before conditioning. Furthermore, whereas onset conditioned responses in the control group remained elevated after 30 extinction trials (presentation of CS alone), onset responses in lesioned animals returned to their preconditioning firing level after approximately 10 extinction trials. These results suggest that the amygdala enables the development of long-latency (US anticipatory) responses and prevents the extinction of short-latency onset responses to threatening stimuli. The findings further suggest that auditory cortex cells may participate differently in explicit and implicit memory networks.

A comparison of corticospinal activation by magnetic coil and electrical stimulation of monkey motor cortex.

The effects of different orientations of a Cadwell round magnetic coil (MC) were compared with each other and with surface electrical stimulation of motor cortex in monkeys anesthetized with pentobarbital or urethane. Recordings were made from within the lateral corticospinal tract, either from axonal populations or with a microelectrode from individual axons. A lateral-sagittally orientated MC directly excited corticospinal neurons at lower stimulus intensity than was required for indirect, i.e., transsynaptic excitation via inputs to corticospinal neurons. By contrast, in 2 out of 3 macaques tested, a vertex-tangential orientation could excite corticospinal neurons indirectly at lower intensities than were required for direct excitation; at higher intensities, direct excitation also occurred. The site of direct corticospinal excitation by a lateral-sagittally orientated MC was inferred by comparing the response variability and latency to MC and surface electrical stimuli. Cathodal stimuli elicited more variable corticospinal population responses and later individual axonal responses than were obtained with anodal stimuli. The variability in response is attributed to interaction between nearby, on-going synaptic bombardment and the stimulus, implying that surface cathodal stimuli directly activate corticospinal neurons at the spike trigger zone (presumably the initial segment). By contrast, the consistency and reduced latency of the corticospinal responses to surface anodal stimuli are attributed to the direct excitation of corticospinal fibers within the white matter. When the stimulus intensity is clearly above threshold, surface anodal and cathodal stimuli can activate corticospinal neurons both directly and indirectly. Direct corticospinal excitation by the MC can resemble the effects of either surface anodal or surface cathodal stimuli. We conclude that the MC can activate corticospinal neurons at the spike trigger zone or their fibers deeper in white matter. The findings in the monkey are used to interpret the effects of different MC orientations in the human.

The diuretic effects of CL 115,129 (d,1-15-deoxy-16-hydroxy-16(alpha/beta)-vinyl-prostaglandin E2) and l-prostaglandin E2 in dogs.

CL 115,129, the corresponding carboxylic acid and major metabolite of CL 115,347 (d,1-15-deoxy-16-hydroxy-16(alpha/beta)-vinyl-prostaglandin E2 methyl ester), a potent orally and transdermally long acting antihypertensive agent, infused at 0.1 microgram/kg/min into the left renal artery of sodium pentobarbital anesthetized beagle dogs increased urinary volume, sodium (Na+), potassium (K+) and chloride (Cl-) excretion of the left kidney 289, 201, 101 and 229%, respectively, over the 30 min vehicle-treated control periods. At 0.3 microgram/kg/min CL 115,129 caused a 475 and 336% increase in urinary volume and Na+, respectively. l-Prostaglandin E2 (l-PGE2) infused at 0.1 microgram/kg/min into the left renal artery increased urinary volume, Na+, K+ and Cl- excretion of the left kidney of anesthetized beagle dogs 416, 234, 112 and 255%, respectively, over the control. Both CL 115,129 and l-PGE2 did not affect the systemic arterial blood pressure or the electrolyte excretion of the contralateral kidney. It is concluded that in contrast to other conventional vasodilators, which may cause severe water and electrolyte retention, CL 115,347, via its metabolite CL 115,129, may cause diuresis and natriuresis in many clinical settings when used as an antihypertensive.