Conditioning-related single unit activity in the frontal cortex of urethane anesthetized rats

Responses of frontal cortex single units to a tone preceding medial forebrain bundle (MFB) stimulation were recorded in urethane anesthetized rats. The animals were implanted with monopolar electrodes for MFB stimulation and, following recovery, stimulation parameters which supported self-stimulation were determined for each rat. Prior to the unit recording experiment, the animals were trained to associate a 2-sec tone with MFB stimulation. Trials were presented at variable intervals. Under urethane anesthesia, single units were isolated and the responses of units to paired and unpaired tones were determined. The results indicate that conditioning-related responses of frontal cortex single units can be recorded in urethane anesthetized rats.     

Anesthetic-dependent excitability changes in the hypothalamic ventromedial nucleus of the rat

In acute experiments in rats anesthetized with urethane, the field potentials, population spike, and unit activity evoked in the hypothalamic ventromedial nucleus (HVM) by amygdaloid stimulation are significantly increased with respect to control when preceded by a conditioning volley at 20- to 100-ms intervals. Under pentobarbital anesthesia, in contrast, the evoked responses were inhibited by the conditioning stimulus for similar interstimulus intervals. In unanesthetized animals chronically implanted with stimulating and recording electrodes, a facilitation of responses by a conditioning stimulus was observed when they were awake or anesthetized with urethane. When the same animals were anesthetized with pentobarbital the HVM evoked response was inhibited by a conditioning pulse. Frequency facilitation and post-tetanic potentiation of HVM responses were markedly enhanced under urethane, whereas in pentobarbital-anesthetized animals inhibition predominated. Picrotoxin reversed the inhibition under pentobarbital to facilitation. These results suggest that the HVM neuron population is under both excitatory and inhibitory influences from the amygdala, the former being predominant in awake and urethane-anesthetized animals and the latter being expressed under pentobarbital anesthesia and is probably mediated by γ-aminobutyric acid.     

Neural substrates for tone-conditioned bradycardia demonstrated with 2-deoxyglucose. I. Activation of auditory nuclei

The 2-deoxyglucose (2-DG) autoradiographic method was used to map the metabolic activity of auditory nuclei before, during and after conditioning. The experiment involved freely behaving rats in a Pavlovian conditioning paradigm in which a 4-5 kHz frequency modulated tone (CS) was paired with aversive electrical stimulation of the midbrain reticular formation (US). The unconditioned response was a rapid decrease in heart rate evoked by the US. Eight groups of rats were subjected to: (1) the tone CS before conditioning; (2) the US alone; (3) the paired CS-US (acquisition); (4) the tone CS after conditioning (extinction); (5) the US prior to the CS (sensitization); (6) the unpaired CS-US (pseudoconditioning); (7) the CS after pseudoconditioning; and (8) no stimulation. The major finding was the differential effect produced by the same tone before and after conditioning. The results showed that: (a) reticular mechanisms interact with incoming acoustic stimuli and modulate the response of auditory nuclei; (b) within each auditory nucleus the region of overlap of the spatial representations of CS and US developed an enhanced metabolic response during conditioning; and (c) the CS representation within the neuronal space of the tonotopic maps in all auditory nuclei, with the exception of the medial geniculate, reflected the learned behavioral value of the CS. The changes revealed by the 2-DG method represent the first anatomical demonstration of the activating effects of reticular sensitization and conditioning on a sensory system. The observations support the conclusion that auditory responses are dependent on the physical as well as on the behavioral parameters of a stimulus.     

Olfactory associative conditioning in infant rats with brain stimulation as reward. I. Neurobehavioral consequences

In Experiment 1, infant rats were implanted with a stimulating electrode in the medial forebrain bundle/lateral hypothalamus (MFB/LH) on postnatal day 12 (PN12). Four to 6 hours later, the pups underwent associative olfactory conditioning, with half of the pups trained with 30 temporal pairings of odor (5 s) and MFB/LH stimulation (200 Hz, 300 ms), and the other half trained with random presentations of odor and MFB/LH stimulation. On PN13, pups were tested for: (1) behavioral preference for the conditioned odor; (2) focal glomerular layer 2-DG uptake to the odor; or (3) mitral/tufted cell single-unit response pattern to the odor. Odor-MFB/LH pairings produced a relative behavioral preference, enhanced focal 2-DG uptake and a modified mitral/tufted cell response pattern to the conditioned odor. Random training resulted in none of these changes. In Experiment 2, PN12 pups were anesthetized with urethane and single-unit responses of mitral/tufted cells to MFB/LH stimulation were examined. MFB/LH stimulation produced a brief suppression of mitral/tufted cell activity followed either by a prolonged excitation (18/30 cells; 8-10 s duration) or a prolonged suppression (12/30 cells; 10-30 s). These results suggest that pairing olfactory nerve input with MFB/LH stimulation modifies subsequent behavioral and physiological responses to olfactory nerve input alone. Furthermore, the prolonged olfactory bulb response to MFB/LH stimulation may be critical in this modification.     

Unit activity of the septo-hippocampal system in classical conditioning with rewarding brain stimulation

Neuronal activity in dentate, hippocampus, septum and medial forebrain bundle (MFB) was recorded with chronically implanted electrodes in rats. Responses to tone were measured before and after differential conditioning reinforced by rewarding brain stimulation. Differential unit response changes were found in hippocampus and MFB. The hippocampal response was enhanced in the early phase to the CS+ and gradually subsided. A similar pattern was obtained in the dentate, with a less remarkable differentiation. In contrast, MFB showed sustained enhancement to the CS+ during the whole period, but not to the CS—. It was proposed that the CS-UCS connection was formed in the MFB after conditioning, and that this connection had an effect that was the activation of this system by the CS+ itself.     

Classical conditioning of tone-signaled bradycardia modifies 2-deoxyglucose uptake patterns in cortex, thalamus, habenula, caudate-putamen and hippocampal formation

The 2-[14C]deoxyglucose (2-DG) autoradiographic method was used to map metabolic activity in all telencephalic and diencephalic structures of the rat brain during and after classical conditioning. A trial was made of a 4-5 KHz frequency modulated tone (CS) paired with midbrain reticular stimulation (US). The unconditioned response was a rapid bradycardia elicited by the US. Alert rats were injected with 2-DG, placed in a sound-proof chamber, and subjected during 90 min to a given treatment: (1) the CS before conditioning, (2) the US alone, (3) the paired CS-US (acquisition), (4) the CS after conditioning (extinction), (5) the US prior to the CS (sensitization), (6) the unpaired CS-US (pseudoconditioning), (7) the CS after pseudoconditioning and (8) no stimulation. The prefrontal cortex showed discrete regions with enhanced 2-DG uptake during conditioning and pseudoconditioning. A columnar organization was well-defined in the posterior parietal cortex of rats subjected to CS-US pairing. The medial thalamus was greatly activated in all groups subjected to reticular stimulation. The dorsomedial nucleus showed its largest activation during conditioning. The lateral habenula and a caudal portion of caudate-putamen showed an overall increase in 2-DG uptake during conditioning. The hippocampal formation showed a specific pattern of metabolic activation during conditioning and after conditioning. A laminar densitometric analysis showed that 2-DG uptake was concentrated in a central band along the sides of the hippocampal fissure which corresponded to the molecular layers. Only this neuropil band of greater metabolic activity showed the learning-related changes. In addition, the hippocampal formation was the only nonauditory structure in the forebrain which clearly responded to the acquired signal value of the tone CS after conditioning. These changes revealed by 2-DG provide a first demonstration of forebrain substrates with localized metabolic alterations related to learning and reticular sensitization.     

Interactions of septal evoked responses to stimulation of the fornix and medial forebrain bundle

Gross and single unit septal evoked response to stimulation of the fornix and medial forebrain bundle (MFB) were studied in anesthetized, acutely prepared rats. Stimulation of the fornix and MFB produced short-latency antidromic and synaptic activation of localized groups of septal target cells. In addition, stimulation of either pathway produced inhibition of spontaneous single cell activity. The interactions of these responses were studied by delivering paired stimuli to the fornix and/or MFB. A prior stimulus to the fornix potentiated the responses of cell groups synaptically activated by subsequent fornical stimulation, but briefly depressed the sunaptic activation of septal cells by subsequent MFB stimulation. A prior stimulus to the MFB slightly potentiated the synaptic activation of septal cells by a subsequent MFB stimulus, but depressed the responses of cell groups synaptically activated by a subsequent stimulus to the fornix. These results were discussed in terms of their implications for septal organization and function.     

Δ-Tetrahydrocannabinol and the hippocampus: Effects on CA1 field potentials in rats

The effects of Δ⁹-tetiahydrocannabinol (THC) on ortho- and antidromically elicited CA1 field potentials were observed in locally anesthetized rats and in rats anesthetized with urethane. THC augmented amplitudes of population EPSP's as well as orthodromic and antidromic population spikes from pyramidal cells in locally anesthetized animals. Latencies to peak amplitude of these responses were increased. Conditioning-test shock experiments revealed that THC also depressed recurrent inhibition probably mediated by basket cells. In animals under urethane anesthesia THC enhanced test responses, but failed to augment population responses to the conditioning stimulus. It was concluded that THC enhanced postsynaptic excitatory processes but attenuated recurrent inhibition. Urethane anesthesia completely blocked the postsynaptic excitatory effect of THC but had little apparent influence on THC's disinhibitory action.     

Δ9-Tetrahydrocannabinol and the hippocampus: Effects on CA1 field potentials in rats

The effects of Δ⁹-tetiahydrocannabinol (THC) on ortho- and antidromically elicited CA1 field potentials were observed in locally anesthetized rats and in rats anesthetized with urethane. THC augmented amplitudes of population EPSP's as well as orthodromic and antidromic population spikes from pyramidal cells in locally anesthetized animals. Latencies to peak amplitude of these responses were increased. Conditioning-test shock experiments revealed that THC also depressed recurrent inhibition probably mediated by basket cells. In animals under urethane anesthesia THC enhanced test responses, but failed to augment population responses to the conditioning stimulus. It was concluded that THC enhanced postsynaptic excitatory processes but attenuated recurrent inhibition. Urethane anesthesia completely blocked the postsynaptic excitatory effect of THC but had little apparent influence on THC's disinhibitory action.     

Classical eyeblink conditioning in decerebrate guinea pigs

A decerebrate guinea pig preparation was used to test the hypothesis that brainstem-cerebellar circuitry is sufficient for classical delay eyeblink conditioning. Delay conditioning was carried out using a tone conditioned stimulus (CS) paired with a co-terminating, periorbital shock unconditioned stimulus (US). Decerebrate animals readily acquired the conditioned response (CR), while pseudoconditioning yielded no signs of learning. When a longer tone CS was used, the learning became slower. These CRs were adaptive and appropriately timed relative to the US. Subsequent CS-alone trials caused extinction of the CR. These characteristics of the eyeblink conditioning were similar to those reported previously in various species, suggesting that the cerebellum and brainstem are sufficient for this type of learning.     

Conditioning-related changes of unit activity in the dorsal lateral geniculate nucleus of urethane-anaesthetized rats

Changes in geniculate unit activity of urethane-anaesthetized and freely moving rats were investigated during conditioning. The conditioned stimulus (CS) was a flash which was paired with an electrical stimulation of the tail as unconditioned stimulus (US). The discharge rates evoked by the CS during forward conditioning were significantly higher in responding units than those evoked by reversal of CS and US (backward conditioning) or by pseudoconditioning. Tail stimulation alone did not cause significant changes in the firing rate of most of the neurons. In 25% of the investigated neurons the facilitation of activity evoked by forward conditioning persisted during an extinction period of more than 15 min. The effect of conditioning on neuronal activity appeared to be comparable in urethane-anaesthetized rats and in freely moving ones which responded to the US with a slight freezing behavior.     

Electrophysiological characteristics of neurons in forebrain regions implicated in self-stimulation of the medial forebrain bundle in the rat

In an attempt to identify neurons likely to play a role in self-stimulation of the medial forebrain bundle (MFB), action potentials of single neurons in the septum and basal forebrain of anesthetized rats were recorded by means of extracellular electrodes. Refractory period estimates were obtained from cells antidromically activated by stimulation of the lateral hypothalamus or ventral tegmental area, and estimates of interelectrode conduction time were obtained from cells that were driven by stimulation of both sites. The results show that some descending MFB axons arising in the medial septum, diagonal band of Broca and neighboring forebrain structures have characteristics comparable to properties of MFB reward neurons inferred from behavioral experiments.     

Auditory cortex lesions prevent the extinction of Pavlovian differential heart rate conditioning to tonal stimuli in rabbits

The present study examined the role of the auditory cortex in the extinction of differentially conditioned heart rate (HR) responses in rabbits. Lesions were placed bilaterally in wither the auditory cortex or the visual cortex. Three days after recovery from surgery, the auditory cortex lesion group and the visual cortex lesion control group were habituated to the tonal conditioned stimuli (CSs), and then given 2 days of Pavlovian differential conditioning (60 trials per day) in which one tone (CS+) was always paired with the unconditioned stimulus and another tone (CS−) was never paired with the uncondition stimulus. Animals that had demonstrated reliable differential conditioning (CS+ response at least 5 beats greater than the CS− response) were placed on an extinction schedule for 7 days. The extinction schedule was identical to the differential conditioning schedule with the exception that shock never followed the CS+. The results of the study indicate that auditory cortex lesions prevent the extinction of differential bradycardia conditioned responses (CRs) to tonal CSs. Whereas the bradycardia responses to the CS+ quickly extinguished in the group that had control lesions in the visual cortex, the auditory cortex lesion group continued to exhibit significantly larger bradycardiac HR CRs to the CS+ relative to the CS− during all 7 days of extinction. These results suggest that the animals in the auditory cortex lesioned group did not inhibit responses to a previously reinforced stimulus (i.e. CS+) as well as animals with control lesions in the visual cortex.     

The action of norepinephrine in the rat hippocampus. IV. The effects of locus coeruleus stimulation on evoked hippocampal unit activity.

Activity of neurons in the hippocampus (HPC) was recorded in awake, freely moving rats. Most cells were inhibited by either a loud auditory stimulus (tone) or by electrical stimulation of the nucleus locus coeruleus (LC). The inhibitory responses to the tone were antagonized by drugs that interfere with central noradrenergic transmission. When LC stimulation was used as the unconditioned stimulus in a classical conditioning paradigm, previously inhibitory responses to the tone were reinstituted. When behaviorally subthreshold LC stimulation preceded a tone which was correlated with food, the existing conditioned response to the tone was potentiated. These data suggest that the generalized inhibitory response of HPC neurons to a tone is modulated by the noradrenergic pathway and that experimental activation of LC can potentiate HPC responses to behaviorally significant conditioned stimuli.     

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.     

Cocaine enhances resistance to extinction of responding for brain-stimulation reward in adult prenatally stressed rats

The present experiment assessed whether prenatal stress (PS) can alter the ability of acute and chronic cocaine administration to increase and decrease the rewarding effectiveness of the medial forebrain bundle (MFB) using intracranial self-stimulation (ICSS), and also whether PS can affect the extinction of the MFB stimulation response. Adult male offspring of female rats that received PS or no PS (nPS) were implanted with MFB stimulating electrodes, and were then tested in ICSS paradigms. In both nPS and PS offspring, acute cocaine injection decreased ICSS thresholds dose-dependently. However, the threshold-lowering effects at any dose were not significantly different between groups. There was also no group-difference in the threshold-elevating effects of chronic cocaine administration. Nevertheless, chronically drug-administered PS rats exhibited a resistance to the extinguishing of the response for brain-stimulation reward when acutely treated with cocaine, as compared to extinction without cocaine treatment. The results suggest that PS may weaken the ability for response inhibition under cocaine loading in male adult offspring.     

Single-cell responsiveness to sensory stimuli in the mediobasal hypothalamus of rats with partial hypothalamic deafferentations and medial forebrain bundle lesions

Neuroendocrine studies have demonstrated that photic, acoustic, and sciatic nerve stimulation produce adrenocortical discharges and these can be blocked by partial hypothalamic deafferentations and medial forebrain bundle (MFB) lesions. The effects of these sensory stimuli on single-cell activity of mediobasal hypothalamic (MBH) neurons were studied in intact male rats and in animals with anterior, anterolateral, and posterolateral hypothalamic deafferentations and bilateral MFB lesions. The data were analyzed as to changes in responsiveness, facilitation: inhibition ratio, patterns of firing, magnitude of response, frequency distribution, and sensory convergence. Photic projections enter the MBH both anteriorly and posteriorly and via the MFB, acoustic afferent fibers utilize the MFB, and the sciatic impulses arrive posteriorly. The MFB has also an inhibitory modulation on the photic, acoustic, and sciatic responses and AHD has the same effect on the latter. MFB lesions reduced most markedly the sensory convergence of the three modalities. The relation of these electrophysiologic findings to neuroendocrine studies on the neural pathways mediating adrenocortical responses are discussed.     

Response of medial septal neurons to the lontophoretic application of glucocorticoids

Neuroendocrine studies have demonstrated that photic, acoustic, and sciatic nerve stimulation produce adrenocortical discharges and these can be blocked by partial hypothalamic deafferentations and medial forebrain bundle (MFB) lesions. The effects of these sensory stimuli on single-cell activity of mediobasal hypothalamic (MBH) neurons were studied in intact male rats and in animals with anterior, anterolateral, and posterolateral hypothalamic deafferentations and bilateral MFB lesions. The data were analyzed as to changes in responsiveness, facilitation: inhibition ratio, patterns of firing, magnitude of response, frequency distribution, and sensory convergence. Photic projections enter the MBH both anteriorly and posteriorly and via the MFB, acoustic afferent fibers utilize the MFB, and the sciatic impulses arrive posteriorly. The MFB has also an inhibitory modulation on the photic, acoustic, and sciatic responses and AHD has the same effect on the latter. MFB lesions reduced most markedly the sensory convergence of the three modalities. The relation of these electrophysiologic findings to neuroendocrine studies on the neural pathways mediating adrenocortical responses are discussed.     

Generation of cortical event-related slow potentials in the rat involves nucleus basalis cholinergic innervation

These experiments were conducted to gather information regarding the role of cholinergic innervation to the cortex in the generation of event-related slow potentials. The effects of unilateral drug treatments or lesions on ipsilateral and contralateral frontal cortex slow potential (SP) responses were examined in rats. The SP responses were recorded with silver-silver chloride electrodes and were generated by a 2 sec light cue which preceded rewarding medial forebrain bundle stimulation. The following approaches were used: microinjection of GABA, procaine or saline into the nucleus basalis magnocellularis; microinjection of atropine or saline subdurally in the SP recording area; electrolytic lesion of the nucleus basalis area; and kainic acid lesion of the nucleus basalis area. The following bilateral measurements were obtained lesion studies: choline acetyltransferase (ChAT) in cortex and hippocampus; serotonin in cortex, hippocampus, striatum and nucleus accumbens; norepinephrine in cortex and hippocampus; dopamine in striatum and nucleus accumbens; and metabolites of serotonin, norepinephrine and dopamine in these areas. The cortical SP responses were reduced on the side ipsilateral to the injections of GABA and procaine into the nucleus basalis, and on the side of the subdural atropine injection. With either type of lesion, the SP responses on the lesioned side were significantly reduced as compared to the non-lesioned side. Reductions in cortical ChAT and other measures were observed ipsilateral to the electrolytic lesion, but only cortical ChAT activity was reduced in the kainic acid-lesioned animals. Thus, pharmacological depression of nucleus basalis neurons, blockade of cholinergic muscarinic receptors in the cortex, and nucleus basalis lesions that reduce cortical choline acetyltransferase activity depress event-related slow potentials in the rat frontal cortex. These results provide evidence that cortical slow potential responses in the rat are dependent upon cholinergic innervation from the nucleus basalis.     

Prolonged unit responses in thalamic reticular, ventral, and posterior nuclei following lateral hypothalamic and midbrain reticular stimulation.

In unanesthetized postcollicular cerveau isole rats, prolonged responses over 10 sec periods following intracranial stimulation (0.2 sec, 100 Hz, 0.5 msec. 600 muA cathodal pulses) were studied with extracellular recordings of 150 nucleus reticularis and 122 ventral and posterior thalamic units. After medial forebrain bundle (MFB) or midbrain reticular (RET) stimulation, most nucleus reticularis units showed significantly decreased firing, and the MFB and RET effects converged on 64%. None was excited. In ventral and posterior thalamus, however, long-lasting unit responses were mixed. MFB and RET stimuli elicited significantly increased firing in 17% and 20% of the units, respectively, and decreased firing in 21% and 16%, respectively. Convergence of MFB- and RET-elicited responses occurred in 25% of the cells, but none showed convergence of opposite responses which might reflect believed motivational differences between the stimuli. Similar stimuli delivered to ventral medial thalamus evoked pronounced excitation of nucleus reticularis discharge, contrasting the MFB and RET effects and confirming previous reports by others. The substantial responses elicited by MFB and RET stimuli in nucleus reticularis fulfill previously proposed criteria for definition of an arousal dimension in terms of single cell activity. The similarity of MFB and RET effects in the posterior nucleus of thalamus raises questions regarding its role in nociception.