Lateral entorhinal cortex lesions impair both egocentric and allocentric object-place associations.

During navigation, landmark processing is critical either for generating an allocentric-based cognitive map or in facilitating egocentric-based strategies. Increasing evidence from manipulation and single-unit recording studies has highlighted the role of the entorhinal cortex in processing landmarks. In particular, the lateral (LEC) and medial (MEC) sub-regions of the entorhinal cortex have been shown to attend to proximal and distal landmarks, respectively. Recent studies have identified a further dissociation in cue processing between the LEC and MEC based on spatial frames of reference. Neurons in the LEC preferentially encode egocentric cues while those in the MEC encode allocentric cues. In this study, we assessed the impact of disrupting the LEC on landmark-based spatial memory in both egocentric and allocentric reference frames. Animals that received excitotoxic lesions of the LEC were significantly impaired, relative to controls, on both egocentric and allocentric versions of an object-place association task. Notably, LEC lesioned animals performed at chance on the egocentric version but above chance on the allocentric version. There was no significant difference in performance between the two groups on an object recognition and spatial T-maze task. Taken together, these results indicate that the LEC plays a role in feature integration more broadly and in specifically processing spatial information within an egocentric reference frame.

Selective lesions of the cholinergic neurons within the posterior pedunculopontine do not alter operant learning or nicotine sensitization.

Cholinergic neurons within the pedunculopontine tegmental nucleus have been implicated in a range of functions, including behavioral state control, attention, and modulation of midbrain and basal ganglia systems. Previous experiments with excitotoxic lesions have found persistent learning impairment and altered response to nicotine following lesion of the posterior component of the PPTg (pPPTg). These effects have been attributed to disrupted input to midbrain dopamine systems, particularly the ventral tegmental area. The pPPTg contains a dense collection of cholinergic neurons and also large numbers of glutamatergic and GABAergic neurons. Because these interdigitated populations of neurons are all susceptible to excitotoxins, the effects of such lesions cannot be attributed to one neuronal population. We wished to assess whether the learning impairments and altered responses to nicotine in excitotoxic PPTg-lesioned rats were due to loss of cholinergic neurons within the pPPTg. Selective depletion of cholinergic pPPTg neurons is achievable with the fusion toxin Dtx-UII, which targets UII receptors expressed only by cholinergic neurons in this region. Rats bearing bilateral lesions of cholinergic pPPTg neurons (>90% ChAT+ neuronal loss) displayed no deficits in the learning or performance of fixed and variable ratio schedules of reinforcement for pellet reward. Separate rats with the same lesions had a normal locomotor response to nicotine and furthermore sensitized to repeated administration of nicotine at the same rate as sham controls. Previously seen changes in these behaviors following excitotoxic pPPTg lesions cannot be attributed solely to loss of cholinergic neurons. These findings indicate that non-cholinergic neurons within the pPPTg are responsible for the learning deficits and altered responses to nicotine seen after excitotoxic lesions. The functions of cholinergic neurons may be related to behavioral state control and attention rather than learning.

Lateral entorhinal cortex is necessary for associative but not nonassociative recognition memory.

The lateral entorhinal cortex (LEC) provides one of the two major input pathways to the hippocampus and has been suggested to process the nonspatial contextual details of episodic memory. Combined with spatial information from the medial entorhinal cortex it is hypothesised that this contextual information is used to form an integrated spatially selective, context-specific response in the hippocampus that underlies episodic memory. Recently, we reported that the LEC is required for recognition of objects that have been experienced in a specific context (Wilson et al. (2013) Hippocampus 23:352-366). Here, we sought to extend this work to assess the role of the LEC in recognition of all associative combinations of objects, places and contexts within an episode. Unlike controls, rats with excitotoxic lesions of the LEC showed no evidence of recognizing familiar combinations of object in place, place in context, or object in place and context. However, LEC lesioned rats showed normal recognition of objects and places independently from each other (nonassociative recognition). Together with our previous findings, these data suggest that the LEC is critical for associative recognition memory and may bind together information relating to objects, places, and contexts needed for episodic memory formation.

Updating of action-outcome associations is prevented by inactivation of the posterior pedunculopontine tegmental nucleus.

The pedunculopontine tegmental nucleus (PPTg) is in a pivotal position between the basal ganglia and brainstem: it is able to influence and regulate all levels of basal ganglia and corticostriatal activity as well as being a key component of brainstem reticular and motor control circuitry. Consistent with its anatomical position, the PPTg has previously been shown to process rapid, salient sensory input, is a target for Parkinson's disease treatments and has been implicated in associative learning. We explicitly investigated the role of the posterior pPPTg (pPPTg) in action-outcome processes, where actions are performed with the goal-directed aim of obtaining an anticipated outcome. We assessed rats' sensitivity to degradation of the contingency between actions (lever pressing) and outcomes (food reward) during either inactivation of pPPTg by microinjection of the GABA agonist muscimol or control infusions of saline. In response to the degradation of contingency between lever press and food reward, saline treated rats rapidly reduced rates of lever pressing whereas muscimol treated rats (pPPTg inactivation) maintained previous lever pressing rates. In contrast, when the contingency between lever press and food reward was unchanged saline and muscimol treated rats maintained their previous rates of lever pressing. This shows that the pPPTg is critically required for updating associations between actions and outcomes, but not in the continued performance of previously learned associations. These results are consistent with a role for the PPTg in 'higher-order' associative learning and are the first to demonstrate a brainstem role in action-outcome learning.

Lateral entorhinal cortex is critical for novel object-context recognition.

Episodic memory incorporates information about specific events or occasions including spatial locations and the contextual features of the environment in which the event took place. It has been modeled in rats using spontaneous exploration of novel configurations of objects, their locations, and the contexts in which they are presented. While we have a detailed understanding of how spatial location is processed in the brain relatively little is known about where the nonspatial contextual components of episodic memory are processed. Initial experiments measured c-fos expression during an object-context recognition (OCR) task to examine which networks within the brain process contextual features of an event. Increased c-fos expression was found in the lateral entorhinal cortex (LEC; a major hippocampal afferent) during OCR relative to control conditions. In a subsequent experiment it was demonstrated that rats with lesions of LEC were unable to recognize object-context associations yet showed normal object recognition and normal context recognition. These data suggest that contextual features of the environment are integrated with object identity in LEC and demonstrate that recognition of such object-context associations requires the LEC. This is consistent with the suggestion that contextual features of an event are processed in LEC and that this information is combined with spatial information from medial entorhinal cortex to form episodic memory in the hippocampus.

Bar pressing for food: differential consequences of lesions to the anterior versus posterior pedunculopontine.

The pedunculopontine tegmental nucleus (PPTg) is in a key position to participate in operant reinforcement via its connections with the corticostriatal architecture and the medial reticular formation. Indeed, previous work has demonstrated that rats bearing lesions of the whole PPTg are impaired when learning to make two bar presses for amphetamine reinforcement. Anterior and posterior portions of the PPTg make different anatomical connections, including preferential projections by the anterior PPTg to substantia nigra pars compacta dopamine neurons and by the posterior PPTg to ventral tegmental area dopamine neurons. We wanted to assess the effects of anterior and posterior PPTg ibotenate lesions on rats learning simple and more complex schedules of natural reinforcement. We trained rats with lesions to the anterior PPTg (n = 11) and the posterior PPTg (n = 5) [and appropriate controls (n = 15)] to bar press for food on a variety of fixed-ratio and variable-ratio reinforcement schedules and then during extinction. We found that posterior PPTg-lesioned rats bar pressed at lower rates, were slower to learn to bar press, and often had deficits characteristic of impaired learning and/or motivation. In contrast, anterior PPTg-lesioned rats learned to bar press for reinforcement at normal rates. However, they made errors of perseveration and anticipation throughout many schedules, and pressed at a higher rate than controls during extinction, deficits best characterized as reflecting disorganized response control. Together, these data suggest that the anterior PPTg and posterior PPTg (and their related circuits) contribute differently to reinforcement learning, incentive motivation, and response control, processes that are considered to malfunction in drug addiction.

Neurons in dopamine-rich areas of the rat medial midbrain predominantly encode the outcome-related rather than behavioural switching properties of conditioned stimuli.

Midbrain dopamine neurons are phasically activated by a variety of sensory stimuli. It has been hypothesized that these activations contribute to reward prediction or behavioural switching. To test the latter hypothesis we recorded from 131 single neurons in the ventral tegmental area and retrorubral field of thirsty rats responding during a modified go/no-go task. One-quarter (n = 33) of these neurons responded to conditioned stimuli in the task, which varied according to the outcome with which they were associated (saccharin or quinine solution) and according to whether they triggered a switch in the ongoing sequence of the animal's behaviour ('behavioural switching'). Almost half the neurons (45%) responded differentially to saccharin- vs. quinine-conditioned stimuli; the activity of a minority (15%) correlated with an aspect of behavioural switching (mostly exhibiting changes from baseline activity in the absence of a behavioural switch) and one-third (33%) encoded various outcome-switch combinations. The strongest response was excitation to the saccharin-conditioned stimulus. Additionally, a proportion (38%) of neurons responded during outcome delivery, typically exhibiting inhibition during saccharin consumption. The neurons sampled did not fall into distinct clusters on the basis of their electrophysiological characteristics. However, most neurons that responded to the outcome-related properties of conditioned stimuli had long action potentials (> 1.2 ms), a reported characteristic of dopamine neurons. Moreover, responses to saccharin-conditioned stimuli were functionally akin to dopamine responses found in the macaque and rat nucleus accumbens responses observed within the same task. In conclusion, our data are more consistent with the reward-prediction than the behavioural switching hypothesis.

Development of a behavioral task measuring reward "wanting" and "liking" in rats.

It has been suggested that reward "wanting" and "liking" are mediated by separable brain systems. To facilitate neuropharmacological and neurophysiological research on this issue we developed a behavioral task with putative measures of reward "wanting" and "liking" available on a trial-by-trial basis. We were able to test whether our measures were sensitive to changes in thirsty rats' "wanting" and "liking" of liquid reward by manipulating its delay, taste and volume. We found that three of our putative "wanting" measures (anticipatory errors, reaction time and reward collection latency) were affected by upcoming reward delay and/or taste and our putative "liking" measure (post-reward licking) was sensitive to variations in reward taste and volume. To cross-validate our measures with previous pharmacological work we tested rats following acute, systemic administration of drug compounds that globally enhance serotonin and noradrenaline (imipramine), dopamine (GBR 12909) and opioid (morphine) function. Imipramine augmented the effects of delay and taste on reward "wanting", GBR 12909 attenuated the effects of delay on reward "wanting" and the effects of taste on reward "liking", and morphine reduced the effect of delay on a measure of reward "wanting". Since morphine failed to affect reward "liking" but has been previously found to enhance reward "liking" in taste reactivity tests, our measure requires further pharmacological validation. However, this task shows potential to assess the specific neural mechanisms that contribute to the impact of reward parameters on "wanting" and "liking".

Rat nucleus accumbens neurons predominantly respond to the outcome-related properties of conditioned stimuli rather than their behavioral-switching properties.

It has been proposed that nucleus accumbens neurons respond to outcome (reward and punishment) and outcome-predictive information. Alternatively, it has been suggested that these neurons respond to salient stimuli, regardless of their outcome-predictive properties, to facilitate a switch in ongoing behavior. We recorded the activity of 82 single-nucleus accumbens neurons in thirsty rats responding within a modified go/no-go task. The task design allowed us to analyze whether neurons responded to conditioned stimuli that predicted rewarding (saccharin) or aversive (quinine) outcomes, and whether the neural responses correlated with behavioral switching. Approximately one third (28/82) of nucleus accumbens neurons exhibited 35 responses to conditioned stimuli. Over 2/3 of these responses encoded the nature of the upcoming rewarding (19/35) or aversive (5/35) outcome. No response was selective solely for the switching of the rat's behavior, although the activity of approximately one third of responses (11/35) predicted the upcoming outcome and was correlated with the presence or absence of a subsequent behavioral switch. Our data suggest a primary functional role for the nucleus accumbens in encoding outcome-predicting information and a more limited role in behavioral switching.

Nucleus accumbens neurons in the rat exhibit differential activity to conditioned reinforcers and primary reinforcers within a second-order schedule of saccharin reinforcement.

The nucleus accumbens has been associated with processing information related to primary reinforcement and reward. Most neurophysiological studies report that nucleus accumbens neurons are phasically excited in response to the onsets of salient events during the seeking of reinforcement and to the delivery of primary reinforcers. However, a minority of studies report inhibition during primary reinforcement. We recorded from 65 neurons in the nucleus accumbens whilst thirsty rats performed under a second-order schedule of saccharin reinforcement. This allowed us to analyse neural activity and behaviour during reinforcer-seeking in the presence of conditioned reinforcers (second-order stimuli, also called 'conditioned stimuli'), and during primary reinforcer consumption. Specifically, we sought to examine the valence of potential neural responses to primary reinforcement, to compare these responses to second-order stimulus-evoked responses, and to determine whether responses were differential to second-order stimuli presented at different time points within the schedule. Fifty out of 65 neurons we sampled responded to the second-order stimulus and/or consumption of the primary reinforcer. Most neurons in our sample exhibited excitation following the second-order stimulus and inhibition to the primary reinforcer, a pattern also present over the average response of the neural population. However, there was no systematic variation in neural responses evoked by second-order stimuli presented at different temporal proximities to primary reinforcement. Our results provide evidence that partially overlapping mechanisms within the nucleus accumbens differentially process conditioned reinforcers and primary reinforcers.

Second-order stimuli do not always increase overall response rates in second-order schedules of reinforcement in the rat.

RATIONALE: Second-order schedules of reinforcement have been used extensively to model reward-seeking and drug-seeking behaviour. Second-order stimuli within second-order schedules have been shown to enhance response rates during operant responding for natural reinforcers and drug reinforcers. This has led some to view second-order schedules of drug reinforcement as a model maintained of drug-seeking in addicts by drug-associated stimuli. However, the functional role of the second-order stimulus within second-order schedules is complex. OBJECTIVE: We investigated the role of second-order stimuli within a second-order schedule of reinforcement [FI 4 min (FR10: S)] maintained by sweetened water reinforcement. METHODS: Eight rats were trained to press a bar on a second-order schedule of reinforcement and tested in the presence and absence of the second-order stimulus. RESULTS: In contrast to most previous work, overall bar-pressing rates were significantly increased when the second-order stimulus was omitted (second-order stimulus omission: 0.17 Hz (+/-0.04, 95% CI); second-order stimulus present: 0.13 Hz (+/-0.04, 95% CI)). However, second-order stimuli also changed the pattern of responding whereby rats would make a bout of bar presses prior to the presentation of the second-order stimulus and then pause briefly after the second-order stimulus. In the absence of second-order stimuli, responding was uniformly high. Control measures, such as the ability of the second-order stimulus to evoke checking for the primary reinforcers, indicated that the second-order stimulus was associated with the primary reinforcer. CONCLUSIONS: These results demonstrated that although second-order stimuli maintained responding and caused the rat to check for primary reinforcement, overall response rates were increased when the second-order stimuli were omitted. This has implications for interpreting the results of studies where overall response rates within second-order schedules have been the only measure used to assess the effects of potential anti-addiction drugs. Future studies could be improved by performing a second-order stimulus omission test analysing both the overall response rates and the temporal organization of responding with respect to the second-order stimulus.