Conditioned cortical slow potential responses in urethane anesthetized rats

Cortical slow potential (SP) responses to tone or light stimuli preceding medial forebrain bundle (MFB) stimulation were recorded in urethane anesthetized rats. In the first study, rats were implanted with Ag-AgCl electrodes for recording frontal cortex SPs as well as monopolar electrodes for MFB stimulation. Following recovery, optimum stimulation parameters for SP conditioning were determined for each rat during self-stimulation sessions. These animals were then subjected to extensive associative conditioning in the unanesthetized state. Trials were presented at variable intervals and a 2-sec tone preceded a single 0.5 sec train of MFB stimulation. Negative SP responses developed with training and responses of similar waveform and amplitude were observed in the same animals under urethane anesthesia. Other rats were implanted with MFB stimulating electrodes and, after recovery, stimulation parameters were determined as above but the animals were not subjected to the conditioning procedure prior to urethane administration. Under urethane anesthesia, Ag-AgCl electrodes were placed on the dura over frontal cortex for recording SP responses during pseudoconditioning, conditioning, extinction and retraining trials, using either light or tone stimuli. Negative bilateral SP responses to the tone or light were minimal or nonexistent during pseudoconditioning, developed gradually with pairing, diminished markedly during extinction and returned to maximum amplitude with retraining. The SP responses also reflected discrimination between reinforced and nonreinforced tone and light stimuli as well as reversal conditioning. Furthermore, turning off a light could also serve as the conditioned stimulus for SP response generation. Cortical slow potential responses can be conditioned in urethane anesthetized rats. Therefore, it may be possible to apply additional neurophysiological techniques in these animals to investigate event-related slow potential mechanisms.     

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.     

Locus coeruleus neurons in the neonatal rat: electrical activity and responses to sensory stimulation

Newborn rats, 1-3 days of age, were anesthetized with urethane and single-unit activities were recorded extracellularly in the locus coeruleus. Ca. 35% of locus coeruleus neurons recorded were antidromically activated from stimulation of the frontal cortex. Although the majority of locus coeruleus neurons were not spontaneously active, non-noxious as well as noxious sensory stimuli such as touches to the skin, air puffs and tail pinches were very effective in exciting locus coeruleus neurons.     

Muscarinic dependence of nucleus basalis induced conditioned receptive field plasticity

Receptive field (RF) plasticity in primary auditory cortex of adult animals, specifically selective increased response to a tonal conditioned stimulus (CS) relative to other frequencies, can be induced both by behavioral conditioning and by pairing a tone with stimulation of the nucleus basalis (NB). This study determined whether cortical muscarinic receptors are necessary for NB-induced RF plasticity. Single units in layers II-IV were studied in Urethane anesthetized adult rats. The cortex was perfused with saline or saline+atropine sulfate. Conditioning, 30 trials of pairing a tone with NB stimulation, produced a significant CS-specific response increase (n=8). Local atropine blocked NB-induced RF plasticity, actually resulting in CS-specific response decrease (n=6). Therefore, NB-induced RF plasticity requires engagement of muscarinic receptors in auditory cortex.     

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.     

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.     

Dorsal raphe stimulation, 5-HT and morphine microiontophoresis effects on noxious and nonnoxious identified neurons in the medial thalamus of the rat

In single cell experiments, the characterization of the responses of medial thalamic neurons to noxious and nonnoxious stimulation was made to examine the effects of two substances involved in pain, morphine and 5-HT, and the action of one pain suppressor mechanism, dorsal raphe stimulation. Single cell activity was recorded in urethane anesthetized rats. Tail pinch and tail immersion in hot water were used as nociceptive stimuli. Skin strokes, air puffs and hair brushing were used as nonnociceptive stimuli. Morphine, 5-HT microiontophoresis and dorsal raphe stimulation were performed in all the recorded units. Fifty-eight percent from 61 medial thalamic recorded units responded both to noxious and nonnoxious stimulation; whereas only 18% and 24.6% of the units responded exclusively to noxious and nonnoxious stimulation, respectively. The noxious responding units were located in the most posterior portions of the medial thalamus. Dorsal raphe stimulation and 5-HT ejection prevented the excitation elicited by noxious input. Morphine ejection prevented both the noxious and nonnoxious input in medial thalamus, in a different population as compared to dorsal raphe stimulation or 5-HT ejection. These findings support the existence of a pain ascending mechanism mediated by an opioid-serotonergic interaction in the medial thalamus of the rat.     

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.     

Ventral pallidum projections to mediodorsal nucleus of the thalamus: an anatomical and electrophysiological investigation in the rat

Horseradish peroxidase (HRP) and single unit recording experiments were done in rats to investigate neural connections from the ventral pallidal region to the mediodorsal nucleus of the thalamus (MD). In the first series, following the diffusion or iontophoretic injection of HRP into the MD, retrogradely labeled neurons were observed throughout the rostrocaudal extent of the ipsilateral ventral pallidum. Most of the labeled neurons were found in an area between the nucleus of the diagonal band and the ventral aspect of the substantia innominata subcommissuralis. Additional labeled neurons were found in the ventral aspect of the globus pallidus and substantia innominata sublenticularis. In the second series, the region shown to contain labeled neurons was explored for single units antidromically activated by single pulse stimulation of the MD in urethane anesthetized rats. One hundred and fifty-nine single units in the subpallidal area were antidromically activated with latencies corresponding to conduction velocities of 0.2-3.9 m/s. A greater percentage of units in the subcommissural region (50.3%) were activated antidromically as compared to the sublenticular region (27.4%). In the third series, the MD was explored for single units which responded orthodromically to stimulation of the ventral pallidum. Fifty-eight percent (40/69) of MD units responded to stimulation of the subcommissural substantia innominata, whereas 90% (72/80) MD units responded to stimulation of the sublenticular substantia innominata. The most frequent type of orthodromic response observed in MD neurons was inhibition with short onset latencies (less than 10 ms). These data provide anatomical and electrophysiological evidence for the existence of direct pathways from the ventral pallidum to the MD and suggest that this projection is part of a corticosubcortical loop through which the frontal cortex with the ventral striatum and pallidum may contribute to motor function.     

Discriminative conditioning-related slow potential and single-unit responses in the frontal cortex of urethane-anesthetized rats

A model is described for obtaining long-term and stable discriminative conditioning-related slow-potential and single-unit responses from the frontal cortex of urethane-anesthetized rats. Responses were recorded and analyzed to reinforced (rewarding medial forebrain bundle stimulation) and non-reinforced tone cues. In the present study, cortical event-related slow potentials provided an adequate index of the level of discriminative conditioning. Single-unit response patterns are described for 57 neurons which demonstrated a discriminative response to either the reinforced or non-reinforced tone cue.     

Responses of preoptic thermosensitive neurons to medial forebrain bundle stimulation

HORI, T., T. KIYOHARA, T. NAKASHIMA AND M. SHIBATA. Responses of preoptic thermosensitive neurons tomedial forebrain bundle stimulation. BRAIN RES. BULL. 8(6) 667–675, 1982.—Single-unit responses of neurons in the preoptic and anterior hypothalamus (PO/AH) to local thermal stimulation and electrical stimulation of the medial forebrain bundle (MFB) were studied in urethane-anesthetized male rats. In a total of 286 units (112 warm-units, 37 cold-units and 137 thermally insensitive units), 109 units (49 warm-units, 13 cold-units and 47 thermally insensitive units) responded to single pulse stimulation of MFB. The units initially inhibited by MFB stimulation corresponded to 64.2% (70 of 109), the units with facilitatory responses were 27.5% (30 of 109) and the antidromically activated units were 8.3% (9 of 109). High incidence of inhibition by noradrenaline (NA) applied iontophoretically was observed in the neurons inhibited by the MFB stimulation. Iontophoretic application of dichloroisoproterenol to 2 warm-units blocked both the NA-induced inhibition and the MFB-induced inhibition. These ascending and descending connections of the MFB with PO/AH thermosensitive neurons may be part of the neural circuits responsible for thermoregulation.     

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.     

Antidromic activation of rat dorsomedial hypothalamic neurons from locus coeruleus and median eminence

In rats anesthetized with urethane, single unit activity was recorded in the hypothalamic dorsomedial nucleus (DMH) to obtain antidromic response to stimulation of locus coeruleus (LC) and median eminence (ME). Ninety-two cells were activated antidromically from LC and/or ME. Antidromic latencies to LC stimulation ranged from 7 to 39 msec and those to ME stimulation ranged from 5 to 20 msec. Approximately 13% of the neurons recorded revealed antidromic responses simultaneously from LC and ME, and they were found to bifurcate near the soma. The majority of DMH neurons projecting to LC alone were not spontaneously active, while those projecting to ME tended to discharge spontaneously.     

Opposite medial thalamic unit responses to rewarding and aversive brain stimulation

Medial thalamus receives fibers from both medial forebrain bundle (MFB) and hindbrain and midbrain reticular formation (RET). The MFB and RET stimulations are rewarding and aversive respectively. In 32 unanesthetized cerveau isolé rats, 158 units were recorded. The MFB and RET effects converge on two thirds of the units recorded in the dorsal medial and paracentral nuclei of thalamus and are opposite in the following ways: post-stimulus pattern of unit discharge during 7 Hz stimulation; slow-wave recruiting with MFB, but not RET, 7 Hz stimulation; and at a “desynchronization” stimulus frequency (20 Hz), MFB elicits decreased unit discharge and RET elicits increased unit discharge, compared to the 7 Hz rates. In the intralaminar and parafasicular nuclei, post-train (60 Hz, 0.2 sec) decreases and increases in unit firing lasting seconds are often elicited with MFB and RET trains respectively. Stimulation of hypothalamic sites outside MFB did not elicit these MFB effects; parafasicular stimulation did not elicit the RET effects. The MFB effects were not seen in habenula or ventral basal thalamus. Hippocampal, habenular, and ventral medial thalamic units did not show opposite MFB and RET effects. Threshold currents for the opposite MFB and RET effects are similar to those eliciting self-stimulation and escape respectively in several operated rats tested both behaviorally and neurophysiologically.     

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.     

The influence of transcortical relationships on visual responses in rats

Visual responses of the 17/18 border of one hemisphere in anesthetized rats were studied while the opposite cortex was depressed. The depression of the opposite cortex was achieved by a cryoblockade of the homotopic area or by bleaching the unstimulated eye. In the first series of experiments field potentials were recorded, whereas in the second experimental series tests were carried out on single units. Bleaching the unstimulated eye produced mostly an enhancement of long latency potentials recorded in deeper layers. Cryoblockade of the contralateral cortex depressed the responses in a more uniform fashion. Most units typically presented a weaker evoked firing after cryoblockade. However, in some neurons the reaction to the treatment depended upon the nature of the stimulus and other units lost their orientation specificity. A single explanation such as a tonic excitatory transcortical influence is insufficient to account for these results.     

Frontal cortex stimulation evoked neostriatal potentials in rats: intracellular and extracellular analysis

Evoked potentials, action potentials and intracellular events were recorded in the neostriatum of urethane anesthetized rats to electrical stimulation of frontal cortex white matter, motor cortex and pre-limbic cortex. Five major waves of the evoked potential were identified. Wave N1 (3.9 msec latency) was small, preceded cellular events and probably represents activation of corticostriate terminals. Wave P1 (10.8 msec latency to peak following white matter stimulation) coincided with an EPSP and neuronal firing. Both wave N2 (38.0 msec latency to peak) and P2 (approximately 110 msec duration) overlapped the intracellularly recorded hyperpolarization and inhibition of cell firing. Based upon this correspondence and upon the behavior of waves N2 and P2 with changing current and during conditioning-test paired pulse stimulation, it was concluded that the waves represent different processes contributing to the cellular hyperpolarization. A late wave, N3 (175 msec onset latency) corresponded to a late rebound firing and cellular depolarization. This late wave was eliminated from the neostriatum, but not from the overlying sensorimotor cortex, by kainic acid lesions that destroyed medial thalamus but left thalamic lateral nuclei and reticular nucleus intact.     

Prefrontal cortex neuron activity during a discriminative conditioning paradigm in unanesthetized rats

Single neuron responses were recorded from the prefrontal cortex of unanesthetized, restrained rats trained to a discriminative conditioning paradigm. The animals were preconditioned to a tone paired with footshock (CS+) and a second tone presented unpaired (CS-). Only neurons with large amplitude, positive first deflection action potentials were studied. Eighty-five percent of the units (17 of 20) emitted conditioned responses in that the response to either one or both of the conditioned stimuli (CS) was significantly different from spontaneous activity. Thirty-five percent of the units emitted differential conditioned responses in that the responses to one of the CS was significantly different than the response to the other CS and that the response was subsequently shown to extinguish when the footshock was withheld. The neurons which emitted the differential conditioned responses appeared to be located in the deeper layers of the anteromedial prefrontal cortex while the remaining neurons were located either in more superficial layers or anterior aspects of the prefrontal cortex. The results suggest that the anteromedial prefrontal cortex may be involved in neural mechanisms related to discriminative conditioning.     

Thermogenetic changes following frontal neocortex stimulation

Heat production changes were recorded in anesthetized female Sprague-Dawley rats after stimulation of orbital frontal neocortex. The results obtained show that orbital frontal neocortex stimulation significantly increases oxygen consumption, and core and brown adipose tissue temperature. The increase was more substantial after stimulation of left than right cortex. Administration of the beta-blocker propranolol abolished the increase in O2 consumption, core and brown adipose tissue temperature following cortical stimulation. These results are in agreement with our previous research showing that functional ablation of cerebral cortex blocked the increase in thermogenesis following lateral hypothalamic lesion. These findings also show that the orbital frontal neocortex in rats is specifically involved in the control of thermogenesis.     

Cerebellar modulation of frontal cortex dopamine efflux in mice: relevance to autism and schizophrenia

Cerebellar and frontal cortical pathologies have been commonly reported in schizophrenia, autism, and other developmental disorders. Whether there is a relationship between prefrontal and cerebellar pathologies is unknown. Using fixed potential amperometry, dopamine (DA) efflux evoked by cerebellar or, dentate nucleus electrical stimulation (50 Hz, 200 muA) was recorded in prefrontal cortex of urethane anesthetized lurcher (Lc/+) mice with 100% loss of cerebellar Purkinje cells and wildtype (+/+) control mice. Cerebellar stimulation with 25 and 100 pulses evoked prefrontal cortex DA efflux in +/+ mice that persisted for 12 and 25 s poststimulation, respectively. In contrast, 25 pulse cerebellar stimulation failed to evoke prefrontal cortex DA efflux in Lc/+ mice indicating a dependency on cerebellar Purkinje cell outputs. Dentate nucleus stimulation (25 pulses) evoked a comparable but briefer (baseline recovery within 7 s) increase in prefrontal cortex DA efflux compared to similar cerebellar stimulation in +/+ mice. However, in Lc/+ mice 25 pulse dentate nucleus evoked prefrontal cortex DA efflux was attenuated by 60% with baseline recovery within 4 s suggesting that dentate nucleus outputs to prefrontal cortex remain partially functional. DA reuptake blockade enhanced 100 pulse stimulation evoked prefrontal cortex responses, while serotonin or norepinephrine reuptake blockade were without effect indicating the specificity of the amperometric recordings to DA. Results provide neurochemical evidence that the cerebellum can modulate DA efflux in the prefrontal cortex. Together, these findings may explain why cerebellar and frontal cortical pathologies co-occur, and may provide a mechanism that accounts for the diversity of symptoms common to multiple developmental disorders.