Visualising pathways for learning about novelty in the rodent brain

Recognition memory is the ability to distinguish novel from familiar stimuli. This thesis explores opposing models of recognition memory that alternatively assume that the perirhinal cortex and hippocampus (regions of the medial temporal lobe) must functionally interact to support recognition memory or that the perirhinal cortex can support this process independently. Additionally, the way in which these areas differentially interact to support learning about novel compared to familiar stimuli was examined. To achieve this, rats with lesions to the hippocampus or perirhinal cortex were given tests of object recognition memory or allowed to explore novel stimuli, after which, regional neuronal activity and network interactions were explored. This was achieved by immediate-early gene imaging; the expression of c-fos was used as a marker of neuronal activity, allowing for the assessment of regional activity at an extremely high anatomical resolution. Network interactions were explored using structural equation modelling; a statistical technique that made it possible to test if the observed activity could be mapped on to known anatomical pathways. In this way, network dynamics supporting these behavioural tasks were explored. Thus, the functional interdependence of the hippocampus and perirhinal cortex was tested both when the brain was intact and following lesions. This was done at multiple levels; behaviourally, at the level of regional activation and at the level of systems interactions

Visualising pathways for learning about novelty in the rodent brain

Recognition memory is the ability to distinguish novel from familiar stimuli. This thesis explores opposing models of recognition memory that alternatively assume that the perirhinal cortex and hippocampus (regions of the medial temporal lobe) must functionally interact to support recognition memory or that the perirhinal cortex can support this process independently. Additionally, the way in which these areas differentially interact to support learning about novel compared to familiar stimuli was examined. To achieve this, rats with lesions to the hippocampus or perirhinal cortex were given tests of object recognition memory or allowed to explore novel stimuli, after which, regional neuronal activity and network interactions were explored. This was achieved by immediate-early gene imaging; the expression of c-fos was used as a marker of neuronal activity, allowing for the assessment of regional activity at an extremely high anatomical resolution. Network interactions were explored using structural equation modelling; a statistical technique that made it possible to test if the observed activity could be mapped on to known anatomical pathways. In this way, network dynamics supporting these behavioural tasks were explored. Thus, the functional interdependence of the hippocampus and perirhinal cortex was tested both when the brain was intact and following lesions. This was done at multiple levels; behaviourally, at the level of regional activation and at the level of systems interactions

Do the rat anterior thalamic nuclei contribute to behavioural flexibility?

The rodent anterior thalamic nuclei (ATN) are vital for spatial memory. A consideration of their extensive frontal connections suggests that these nuclei may also subserve non-spatial functions. The current experiments explored the importance of the ATN for different aspects of behavioural flexibility, including their contribution to tasks typically associated with frontal cortex. In Experiment 1, rats with ATN lesions were tested on a series of response and visual discriminations in an operant box and, subsequently, in a water tank. The tasks included assessments of reversal learning as well switches between each discrimination dimension. Results revealed a mild and transient deficit on the operant task that was not specific to any stage of the procedure. In the water tank, the lesion animals were impaired on the reversal of a spatial discrimination but did not differ from controls on any other measure. Experiment 2 examined the impact of ATN damage on a rodent analogue of the ‘Stroop’, which assesses response choice during stimulus conflict. The lesion animals successfully acquired this task and were able to use contextual information to disambiguate conflicting cue information. However, responding during the initial presentation of conflicting cue information was affected by the lesion. Taken together, these results suggest that the ATN are not required for aspects of behavioural flexibility (discrimination learning, reversals or high-order switches) typically associated with the rat medial prefrontal cortex. The results from Experiment 2 suggest that the non-spatial functions of the ATN may be more aligned with those of the anterior cingulate cortex

Detecting and discriminating novel objects : The impact of perirhinal cortex disconnection on hippocampal activity patterns

Funded by Wellcome Trust. Grant Numbers: , WT09520Peer reviewedPublisher PD

Collateral projections innervate the mammillary bodies and retrosplenial cortex: A new category of hippocampal cells

To understand the hippocampus it is necessary to understand the subiculum. Unlike other hippocampal subfields, the subiculum projects to almost all distal hippocampal targets, highlighting its critical importance for external networks. The present studies, in male rats and mice, reveal a new category of dorsal subiculum neurons that innervate both the mammillary bodies and the retrosplenial cortex. These bifurcating neurons comprise almost half of the hippocampal cells that project to retrosplenial cortex. The termination of these numerous collateral projections was visualized within the medial mammillary nucleus and the granular retrosplenial cortex (area 29). These collateral projections included subiculum efferents that cross to the contralateral mammillary bodies. Within the granular retrosplenial cortex, the collateral projections form a particularly dense plexus in deep layer II and layer III. This retrosplenial termination site co-localized with markers for VGluT2 and neurotensin. While efferents from the hippocampal CA fields standardly collateralize, subiculum projections often have only one target site. Consequently, the many collateral projections involving the retrosplenial cortex and the mammillary bodies present a relatively unusual pattern for the subiculum, which presumably relates to how both targets have complementary roles in spatial processing. Furthermore, along with the anterior thalamic nuclei, the mammillary bodies and retrosplenial cortex are key members of a memory circuit, which is usually described as both starting and finishing in the hippocampus. The present findings reveal how the hippocampus simultaneously engages different parts of this circuit, so forcing an important revision of this networ

Perirhinal cortex lesions in rats: Novelty detection and sensitivity to interference

Rats with perirhinal cortex lesions received multiple object recognition trials within a continuous session to examine whether they show false memories. Experiment 1 focused on exploration patterns during the first object recognition test postsurgery, in which each trial contained 1 novel and 1 familiar object. The perirhinal cortex lesions reduced time spent exploring novel objects, but did not affect overall time spent exploring the test objects (novel plus familiar). Replications with subsequent cohorts of rats (Experiments 2, 3, 4.1) repeated this pattern of results. When all recognition memory data were combined (Experiments 1–4), giving totals of 44 perirhinal lesion rats and 40 surgical sham controls, the perirhinal cortex lesions caused a marginal reduction in total exploration time. That decrease in time with novel objects was often compensated by increased exploration of familiar objects. Experiment 4 also assessed the impact of proactive interference on recognition memory. Evidence emerged that prior object experience could additionally impair recognition performance in rats with perirhinal cortex lesions. Experiment 5 examined exploration levels when rats were just given pairs of novel objects to explore. Despite their perirhinal cortex lesions, exploration levels were comparable with those of control rats. While the results of Experiment 4 support the notion that perirhinal lesions can increase sensitivity to proactive interference, the overall findings question whether rats lacking a perirhinal cortex typically behave as if novel objects are familiar, that is, show false recognition. Rather, the rats retain a signal of novelty but struggle to discriminate the identity of that signal

Asymmetric cross-hemispheric connections link the rat anterior thalamic nuclei with the cortex and hippocampal formation

Dense reciprocal connections link the rat anterior thalamic nuclei with the prelimbic, anterior cingulate and retrosplenial cortices, as well as with the subiculum and postsubiculum. The present study compared the ipsilateral thalamic-cortical connections with the corresponding crossed, contralateral connections between these same sets of regions. All efferents from the anteromedial thalamic nucleus to the cortex, as well as those to the subiculum, remained ipsilateral. In contrast, all of these target sites provided reciprocal, bilateral projections to the anteromedial nucleus. While the anteroventral thalamic nucleus often shared this same asymmetric pattern of cortical connections, it received relatively fewer crossed inputs than the anteromedial nucleus. This difference was most marked for the anterior cingulate projections, as those to the anteroventral nucleus remained almost entirely ipsilateral. Unlike the anteromedial nucleus, the anteroventral nucleus also appeared to provide a restricted, crossed projection to the contralateral retrosplenial cortex. Meanwhile, the closely related laterodorsal thalamic nucleus had almost exclusively ipsilateral efferent and afferent cortical connections. Likewise, within the hippocampus, the postsubiculum seemingly had only ipsilateral efferent and afferent connections with the anterior thalamic and laterodorsal nuclei. While the bilateral cortical projections to the anterior thalamic nuclei originated predominantly from layer VI, the accompanying sparse projections from layer V largely gave rise to ipsilateral thalamic inputs. In testing a potentially unifying principle of anterior thalamic – cortical interactions, a slightly more individual pattern emerged that reinforces other evidence of functional differences within the anterior thalamic and also helps to explain the consequences of unilateral interventions involving these nuclei

Advances in the behavioural testing and network imaging of rodent recognition memory

Research into object recognition memory has been galvanised by the introduction of spontaneous preference tests for rodents. The standard task, however, contains a number of inherent shortcomings that reduce its power. Particular issues include the problem that individual trials are time consuming, so limiting the total number of trials in any condition. In addition, the spontaneous nature of the behaviour and the variability between test objects add unwanted noise. To combat these issues, the ‘bow-tie maze’ was introduced. Although still based on the spontaneous preference of novel over familiar stimuli, the ability to give multiple trials within a session without handling the rodents, as well as using the same objects as both novel and familiar samples on different trials, overcomes key limitations in the standard task. Giving multiple trials within a single session also creates new opportunities for functional imaging of object recognition memory. A series of studies are described that examine the expression of the immediate-early gene, c-fos. Object recognition memory is associated with increases in perirhinal cortex and area Te2 c-fos activity. When rats explore novel objects the pathway from the perirhinal cortex to lateral entorhinal cortex, and then to the dentate gyrus and CA3, is engaged. In contrast, when familiar objects are explored the pathway from the perirhinal cortex to lateral entorhinal cortex, and then to CA1, takes precedence. The switch to the perforant pathway (novel stimuli) from the temporoammonic pathway (familiar stimuli) may assist the enhanced associative learning promoted by novel stimuli

Contrasting networks for recognition memory and recency memory revealed by immediate-early gene imaging in the rat.

The expression of the immediate-early gene c-fos was used to compare networks of activity associated with recency memory (temporal order memory) and recognition memory. In Experiment 1, rats were first familiarized with sets of objects and then given pairs of different, familiar objects to explore. For the recency test group, each object in a pair was separated by 110 min in the time between their previous presentations. For the recency control test, each object in a pair was separated by less than a 1 min between their prior presentations. Temporal discrimination of the objects correlated with c-fos activity in the recency test group in several sites, including area Te2, the perirhinal cortex, lateral entorhinal cortex, as well as the dentate gyrus, hippocampal fields CA3 and CA1. For both the test and control conditions, network models were derived using structural equation modeling. The recency test model emphasized serial connections from the perirhinal cortex to lateral entorhinal cortex and then to the CA1 subfield. The recency control condition involved more parallel pathways, but again highlighted CA1 within the hippocampus. Both models contrasted with those derived from tests of object recognition (Experiment 2), because stimulus novelty was associated with pathways from the perirhinal cortex to lateral entorhinal cortex that then involved both the dentate gyrus (and CA3) and CA1 in parallel. The present findings implicate CA1 for the processing of familiar stimuli, including recency discriminations, while the dentate gyrus and CA3 pathways are recruited when the perirhinal cortex signals novel stimuli