Automated touchscreen systems have become increasingly prevalent in rodent model screening. This technology has significantly enhanced cognitive and behavioral assessments in mice and has bridged the translational gap between basic research using rodent models and human clinical research. Our study introduces a custom-built touchscreen operant conditioning chamber powered by a Raspberry Pi and a commercially available computer tablet, which effectively addresses the significant cost barriers traditionally associated with this technology. In order to test our prototype, we decided to train C57BL/6 mice on a visual discrimination serial-reversal task, and both C57BL/6 and AppNL−G−Fstrain - an Alzheimer’s Disease (AD) mouse model - on a new location discrimination serial-reversal task. The results demonstrated a clear progression toward asymptotic performance, particularly in the location discrimination task, which also revealed potential genotype-specific deficits, with AppNL−G−F mice displaying an increase in the average number of errors in the first reversal as well as in perseverative errors, compared to wild-type mice. These results validate the practical utility of our touchscreen apparatus and underline its potential to provide insights into the behavioral and cognitive markers of neurobiological disorders.

Aging is associated with cognitive decline, and currently, there are no approved medications that can prevent these impairments. Recently, cannabinoids derived from Cannabis sativa have emerged as promising therapeutic compounds with neuroprotective, anti-inflammatory, and cognitive-enhancing properties. Despite their benefits, further research is needed to fully understand their efficacy across various conditions. This study investigates the effects of cannabidiol (CBD) on memory impairment and brain inflammation in aging mice. Fourteen-month-old C57 mice were administered CBD orally for 7 months and subsequently evaluated between 19 and 21 months of age using behavioral tasks that are sensitive to dysfunction of the perirhinal cortex, hippocampus, amygdala, and various brain regions that are crucial for motor control and coordination. The findings of this study indicate that CBD reduces inflammatory response in the brain and improves cognitive decline associated with aging.

Evidence from neurophysiological and genetic studies demonstrates that activity sparsity-the proportion of neurons that are active at a given time in a population-systematically varies across the canonical trisynaptic circuit of the hippocampus. Recent work has also shown that sparsity varies across the hippocampal dorsoventral (long) axis, wherein activity is sparser in ventral than dorsal regions. While the hippocampus has a critical role in long-term memory (LTM), whether sparsity across the trisynaptic circuit and hippocampal long axis is task-dependent or invariant remains unknown. Importantly, representational sparsity has significant implications for neural computation and theoretical models of learning and memory within and beyond the hippocampus. Here we used functional molecular imaging to quantify sparsity in the rat hippocampus during performance of the Morris water task (MWT) and contextual fear discrimination (CFD) - two popular and distinct assays of LTM. We found that activity sparsity is highly reliable across memory tasks, wherein activity increases sequentially across the trisynaptic circuit (DG < CA3 < CA1) and decreases across the long axis (ventral<dorsal). These results have important implications for models of hippocampal function and suggest that activity sparsity is a preserved property in the hippocampal system across cognitive settings.

Chronic inflammatory diseases are frequently comorbid with depression and anxiety, often persisting during periods of inflammatory remission. This suggests functional changes to neural circuits involved in the contextual regulation of motivation and threat processing. Here, we test how chronic gut inflammation evoked by dextran sodium sulfate (DSS) affects gene expression in several limbic brain structures associated with these functions. We assessed post-mortem expression of mRNA transcripts in the anterior cingulate cortex (ACC), CA1 hippocampus, nucleus accumbens (NAc), and primary motor cortex (M1) as a non-limbic control. The levels of mRNA associated with mitochondrial function, inflammation, and synaptic connectivity were altered in DSS-treated animals, but the specific pattern of changes was heterogeneous among brain structures. Chronic gut inflammation affected transcript expression in the CA1 and NAc more so than in the ACC and M1. These differences involved genes related to antioxidant systems and mitochondrial function. For example, expression of the cytochrome oxidase 1 gene mt-co1, which is necessary for oxidative phosphorylation, was reduced in ACC and NAc of DSS animals, suggesting reduced capacity for ATP production in these regions. Markers of gut inflammation correlated with expression of several transcripts in the ACC, including markers of synapses and GABA synthesis. The NAc showed strong correlations of mitochondrial function and measures of mitochondrial fission, inflammation, synaptic connectivity, and GABA synthesis. In sum, the effects of chronic relapsing gut inflammation on mitochondrial and antioxidant transcriptional programs were heterogeneous across key limbic brain structures.

Lameness is a disruption of normal gait due to infection, injury, or abnormal conformation, causing pain and discomfort. Several scoring systems have been developed to determine lameness severity; a key indicator in deciding if an animal is fit to withstand transport. However, most scoring systems were developed for dairy cattle and may not be suited for use in feedlot cattle due to the large genetic variation between beef breeds as well as the highly variable environment in which they are housed. Lack of consistency when scoring lameness and reduced ability to detect early lame cattle can result in poor animal welfare and increased costs. The objectives of this study were 1) to develop a mobility scoring system that aids in transport decisions for beef cattle, and 2) to determine observer repeatability using this scoring system. Over a 7-mo period, video cameras were placed in an alley to record the ambulation of individual cattle exiting the processing barn at 4 commercial feedlots in Southern Alberta. Cameras were set to record whole-body videos of cattle walking, running, or trotting. Five researchers trained to use this mobility scoring system, individually evaluated 174 cattle from the collected videos, for fitness for transport and the presence/absence of gait abnormalities: shortness of stride, stiffness (mild, moderate, or severe), limp (mild, moderate, or severe), hip hike, head dropped, head bob, arched back, non-weight bearing, or reluctance to move. The mobility scoring system used was a modified (NCBA and Zinpro) 4-point scale (0-3) for locomotion: 0) an animal with no gait abnormalities; 1) an animal with mild stiffness and shortness of stride; 2) an animal with difficulty taking some steps and one or more of the following: severe stiffness, limp favoring affected limb, limps with a head bob when walking but still bears weight; and 3) an animal reluctant or unable to move, bearing little or no weight on affected limb, the head of the animal is dropped and back arched with pronounced head bob and severe limp detected, or incoordination. Fitness for transport was scored as fit, compromised, or unfit according to the humane transportation guidelines of the Canadian Food Inspection Agency. A kappa statistic was calculated to assess inter-observer reliability for both mobility and transport fitness scoring systems as well as gait abnormalities. The combined mobility scores had substantial agreement between observers (Κ = 0.63; P < 0.001). Mobility score of 2 had the highest agreement among all variables (Κ = 0.88; P < 0.001), followed by head bob (Κ = 0.85; P < 0.001), and arched back (Κ = 0.76; P < 0.001). Results of this study will aid in the development of a mobility score that improves industry stakeholder agreement regarding lameness severity and transport loading decisions.

Background: The goal of these experiments was to determine which learning and memory system(s) were necessary for the retention of visual discriminations and subsequent acquisition of a second problem. The dorsal striatum should be involved in the acquisition and expression of this task based on previous work implicating this region in instrumental learning and memory processes. The perirhinal cortex has been implicated in learning and memory processes associated with visual information like objects, and pictures and may also play a role in the acquisition and/or retention of visual discriminations. As there is no clear spatial/relational component to the task, the hippocampus should not be involved. Methods: Rats were trained on a two-choice visual discrimination task to criterion performance after which they received lesions to portions of the dorsal striatum (dorso-medial or dorso-lateral striatum) and medial temporal lobe (perirhinal cortex or hippocampus). After surgical recovery, the rats were tested for retention of the original discrimination, followed by training on a second problem on the same task. Results: The results showed that dorsal medial striatal lesions produced a retrograde deficit on picture discrimination, but dorsal lateral striatum lesions did not. Neither dorsal striatal lesion produced a deficit on acquisition of a second problem. Perirhinal cortex did not seem to make an essential contribution to the retention of the original discrimination or acquisition of the second problem. Surprisingly, subjects with hippocampal damage were severely impaired but eventually re-learned the discrimination. Damage to the hippocampus had no impact on acquisition of a second problem. Conclusions: Taken together, the results of the present experiments show that the dorsomedial striatum and the hippocampus may support performance on this instrumental task if intact during acquisition but is not required for acquisition of a new problem. The implications of this pattern of results for our understanding of the organization of learning and memory in mammals is discussed.

Education, occupation, and an active lifestyle, comprising enhanced social, physical, and mental components are associated with improved cognitive functions in aged people and may delay the progression of various neurodegenerative diseases including Alzheimer's disease. To investigate this protective effect, 3-month-old APPNL-G-F/NL-G-F mice were exposed to repeated single- or multi-domain cognitive training. Cognitive training was given at the age of 3, 6, & 9 months. Single-domain cognitive training was limited to a spatial navigation task. Multi-domain cognitive training consisted of a spatial navigation task, object recognition, and fear conditioning. At the age of 12 months, behavioral tests were completed for all groups. Then, mice were sacrificed, and their brains were assessed for pathology. APPNL-G-F/NL-G-F mice given multi-domain cognitive training compared to APPNL-G-F/NL-G-F control group showed an improvement in cognitive functions, reductions in amyloid load and microgliosis, and a preservation of cholinergic function. Additionally, multi-domain cognitive training improved anxiety in APPNL-G-F/NL-G-F mice as evidenced by measuring thigmotaxis behavior in the Morris water maze. There were mild reductions in microgliosis in the brain of APPNL-G-F/NL-G-F mice with single-domain cognitive training. These findings provide causal evidence for the potential of certain forms of cognitive training to mitigate the cognitive deficits in Alzheimer disease.

Damage to the hippocampus produces profound retrograde amnesia, but odour and object discrimination memories can be spared in the retrograde direction. Prior lesion studies testing retrograde amnesia for object/odour discriminations are problematic due to sparing of large parts of the hippocampus, which may support memory recall, and/or the presence of uncontrolled, distinctive odours that may support object discrimination. To address these issues, we used a simple object discrimination test to assess memory in male rats. Two visually distinct objects, paired with distinct odour cues, were presented. One object was associated with a reward. Following training, neurotoxic hippocampal lesions were made using N-methyl-D-aspartate (NMDA). The rats were then tested on the preoperatively learned object discrimination problem, with and without the availability of odour or visual cues during testing. The rats were also postoperatively trained on a new object discrimination problem. Lesion sizes ranged from 67% to 97% of the hippocampus (average of 87%). On the preoperatively learned discrimination problem, the rats with hippocampal lesions showed preserved object discrimination memory when tested in the dark (i.e., without visual cues) but not when the explicit odour cues were removed from the objects. Hippocampal lesions increased the number of trials required to reach criterion but did not prevent rats from solving the postoperatively learned discrimination problem. Our results support the idea that long-term memories for odours, unlike recall of visual properties of objects, do not depend on the hippocampus in rats, consistent with previous observations that hippocampal damage does not cause retrograde amnesia for odour memories.

This study investigated the impact of familial Alzheimer's disease (AD)-linked amyloid precursor protein (App) mutations on hippocampal CA1 neuronal activity and function at an early disease stage in AppNL-G-F/NL-G-F × Thy1-GCaMP6s+/- (A-TG) mice using calcium imaging. Longitudinal assessment of spatial behavior at 12 and 18 months of age identified an early disease stage at 12 months when there was significant amyloid beta pathology with mild behavioral deficits. Hippocampal CA1 neuronal activity and event-related encoding of distance and time were therefore assessed at 12 months of age in several configurations of an air-induced running task to assess the dynamics of cellular activity. Neurons in A-TG mice displayed diminished (weaker) and more frequent (hyperactive) neuronal firing that was more pronounced during movement compared to immobility. Responsive neurons showed configuration-specific deficits in distance and time encoding with impairment in adapting their responses to changing configurations. These results suggest that at an early stage of AD in the absence of full-blown behavioral deficits, weak-hyperactive neuronal activity may induce impairments in sensory perception of changing environments.

BACKGROUND:An active lifestyle is associated with improved cognitive functions in aged people and may prevent or slow down the progression of various neurodegenerative diseases including Alzheimer's disease (AD). To investigate these protective effects, male APPNL-G-F mice were exposed to long-term voluntary exercise.METHODS:Three-month-old AD mice were housed in a cage supplemented with a running wheel for 9 months for long-term exercise. At the age of 12 months, behavioral tests were completed for all groups. After completing behavioral testing, their brains were assessed for amyloid pathology, microgliosis, and cholinergic cells.RESULTS:The results showed that APPNL-G-F mice allowed to voluntarily exercise showed an improvement in cognitive functions. Furthermore, long-term exercise also improved anxiety in APPNL-G-F mice as assessed by measuring thigmotaxis in the Morris water task. We also found reductions in amyloid load and microgliosis, and a preservation of cholinergic cells in the brain of APPNL-G-F mice allowed to exercise in their home cages. These profound reductions in brain pathology associated with AD are likely responsible for the observed improvement of learning and memory functions following extensive and regular exercise.CONCLUSION:These findings suggest the potential of physical exercise to mitigate the cognitive deficits in AD.

ABSTRACT In contrast to most transgenic mouse models of Alzheimer disease (AD), knock-in mice expressing familial AD-linked mutations of the amyloid precursor protein ( App ) gene exhibit stereotypical age-dependent amyloid beta (Aβ) pathology and cognitive impairment without physiologically unrealistic App overexpression. This study investigated the effect of familial AD-linked App mutations on hippocampal CA1 neuronal activity and function. To enable calcium imaging of neuronal activity, App NL-G-F/NL-G-F knock-in (APPki) mice were crossed with Thy1 -GCaMP6s +/- (C-TG) mice to generate App NL-G-F/NL-G-F × Thy1 -GCaMP6s +/- (A-TG) mice, which were characterized at 12 months of age. A-TG mice exhibited Aβ pathology in the hippocampus. In several configurations of an air-induced running task, A-TG mice and C-TG mice were equally successful in learning to run or to stay immobile. In the Morris water place test, A-TG mice were impaired, but learned the task. Comparisons of hippocampal CA1 neuronal activity in the air-induced running task showed that A-TG mice displayed neuronal hypoactivity both during movement and immobility. A-TG mice and C-TG CA1 neuronal encoding of distance or time in the air induced running task were not different. These results suggest that knock-in of familial AD-linked mutations in A-TG mice results in Aβ pathology, neuronal hypoactivity, and cognitive impairment without severely affecting CA1 neuronal encoding. In comparison to APPki mice, A-TG mice had less severe AD-like memory impairments at 12 months of age (Saito et al., 2014; Mehla et al., 2019), suggesting that the disease onset was delayed in A-TG mice. The effect of APP mutations may have been mitigated through genetic mechanisms when APPKi mice were crossed with C-TG mice.

Evidence from genetic, behavioural, anatomical, and physiological study suggests that the hippocampus functionally differs across its longitudinal (dorsoventral or septotemporal) axis. Although, how to best characterize functional and representational differences in the hippocampus across its long axis remains unclear. While some suggest that the hippocampus can be divided into dorsal and ventral subregions that support distinct cognitive functions, others posit that these regions vary in their granularity of representation, wherein spatial-temporal resolution decreases in the ventral (temporal) direction. Importantly, the cognitive and granular hypotheses also make distinct predictions on cellular recruitment dynamics under conditions when animals perform tasks with qualitatively different cognitive-behavioural demands. One interpretation of the cognitive function account implies that dorsal and ventral cellular recruitment differs depending on relevant behavioural demands, while the granularity account suggests similar recruitment dynamics regardless of the nature of the task performed. Here, we quantified cellular recruitment with the immediate early gene (IEG) Arc across the entire longitudinal CA1 axis in female and male rats performing spatial-and fear-guided memory tasks. Our results show that recruitment is greater in dorsal than ventral CA1 regardless of task or sex, and thus support a granular view of hippocampal function across the long axis. We further discuss how future experiments might determine the relative contributions of cognitive function and granularity of representation to neuronal activity dynamics in hippocampal circuits.

Alzheimer’s disease (AD) is characterized neuropathologically by amyloid-β (Aβ) plaques and neurofibrillary tangles. Vascular pathology caused by chronic cerebral hypoperfusion (HP) is hypothesised to exacerbate AD pathology and has emerged as an increasing cause of age-related cognitive impairment. In this study we examined the effects of gradual cerebral HP on cognitive dysfunction, Aβ pathology, microgliosis, and cortical network dynamics in C57BL/6J mice and a single App knock-in mouse model of AD ( App NL-G-F ). We performed unilateral common carotid artery gradual occlusion (UCAgO) in two-month-old mice using an ameroid constrictor. At 4 months of age, animals were tested in a behavioral battery consisting of tests of spatial learning and memory (Morris water task), recognition memory (novel object recognition task), and motor coordination (balance beam). Following behavioural testing, in vivo mesoscale wide-field voltage imaging was done to assess cortical functional connectivity and sensory-evoked cortical activity, and brains were harvested for pathology characterization using immunohistochemistry. We found that UCAgO reduced cerebral blood flow (CBF) in the occluded hemisphere (OH), however, subtle behavioural deficits were observed due to HP. A dissociative effect of HP was observed in resting-state functional connectivity analysis, where HP led to hyper-connectivity in C57 mice and hypo-connectivity in App mice. Interestingly, sensory stimulation of limbs contralateral to OH revealed hyper-cortical activations in the non-occluded hemisphere of C57 HP mice, however, hypo-cortical activations were observed in App HP mice. Furthermore, we found that the UCAgO increased cortical and hippocampal microgliosis in both hemispheres of C57 and App mice, a bilateral increase in Aβ deposition was only observed in App mice. These results suggest that gradual cerebral HP leads to cortical network alterations in AD, which is partly mediated via activation of microglia.

In contrast to most transgenic mouse models of Alzheimer disease (AD), knock-in mice expressing familial AD-linked mutations of the amyloid precursor protein ( App ) gene exhibit stereotypical age-dependent amyloid beta (Aβ) pathology and cognitive impairment without physiologically unrealistic App overexpression. This study investigated the effect of familial AD-linked App mutations on hippocampal CA1 neuronal activity and function. To enable calcium imaging of neuronal activity, AppNL-G-F/NL-G-F knock-in (APPki) mice were crossed with Thy1 -GCaMP6s+/- (C-TG) mice to generate AppNL-G-F/NL-G-F × Thy1 -GCaMP6s+/- (A-TG) mice, which were characterized at 12 months of age. A-TG mice exhibited Aβ pathology in the hippocampus. In several configurations of an air-induced running task, A-TG mice and C-TG mice were equally successful in learning to run or to stay immobile. In the Morris water place test, A-TG mice were impaired, but learned the task. Comparisons of hippocampal CA1 neuronal activity in the air-induced running task showed that A-TG mice displayed neuronal hypoactivity both during movement and immobility. A-TG mice and C-TG CA1 neuronal encoding of distance or time in the air induced running task were not different. These results suggest that knock-in of familial AD-linked mutations in A-TG mice results in Aβ pathology, neuronal hypoactivity, and cognitive impairment without severely affecting CA1 neuronal encoding. In comparison to APPki mice, A-TG mice had less severe AD-like memory impairments at 12 months of age ([Saito et al., 2014][1]; [Mehla et al., 2019][2]), suggesting that the disease onset was delayed in A-TG mice. The effect of APP mutations may have been mitigated through genetic mechanisms when APPKi mice were crossed with C-TG mice.### Competing Interest StatementThe authors have declared no competing interest. [1]: #ref-27 [2]: #ref-24

Documenting a mouse's "real world" behavior in the "small world" of a laboratory cage with continuous video recordings offers insights into phenotypical expression of mouse genotypes, development and aging, and neurological disease. Nevertheless, there are challenges in the design of a small world, the behavior selected for analysis, and the form of the analysis used. Here we offer insights into small world analyses by describing how acute behavioral procedures can guide continuous behavioral methodology. We show how algorithms can identify behavioral acts including walking and rearing, circadian patterns of action including sleep duration and waking activity, and the organization of patterns of movement into home base activity and excursions, and how they are altered with aging. We additionally describe how specific tests can be incorporated within a mouse's living arrangement. We emphasize how machine learning can condense and organize continuous activity that extends over extended periods of time.

Abstract Background. Alzheimer’s disease (AD) is characterized by the prion-like propagation of amyloid-β (Aβ). However, the role of Ab in cognitive impairment is still unclear. Methods. Two-month-old AppNL-G-F were intracerebrally seeded with three conformationally distinct Aβ seeds. The mice were trained in a multi-model paradigm using multiple behavioural tests. Immunohistochemical techniques were used to assess Ab deposition, microgliosis, and cholinergic tone. Results. Seeding accelerated AD pathology throughout the brain in a region-dependent manner. Microgliosis was elevated in the substantia nigra reticular area. The cholinergic tone was found to be significantly reduced in the medial septal band. No impairment in learning and memory was found. Conclusion. These results suggest seeding accelerates microglial and Ab plaque progression in a regional distinct manner with the seed having no unique effect on behaviour or histological markers.

Alzheimer's disease (AD) is characterized by the prion-like propagation of amyloid-β (Aβ). However, the role of Aβ in cognitive impairment is still unclear. To determine the causal role of Aβ in AD, we intracerebrally seeded the entorhinal cortex of a 2-month-old AppNL−G−F mouse model with an Aβ peptide derived from patients who died from rapidly progressing AD. When the mice were 3 months of age or 1 month following seeding, spatial learning and memory were tested using the Morris water task. Immunohistochemical labeling showed seeding with the Aβ was found accelerate Aβ plaque deposition and microgliosis in the AppNL−G−F mice, but this was dependent on the presence of the knocked-in genes. However, we found no correlation between pathology and spatial performance. The results of the present study show the seeding effects in the AppNL−G−F knock-in model, and how these are dependent on the presence of a humanized App gene. But these pathological changes were not initially causal in memory impairment.

Damage to the hippocampus (HPC) typically causes retrograde amnesia for contextual fear conditioning. Repeating the conditioning over several sessions, however, can eliminate the retrograde amnesic effects. This form of reinstatement thus permits modifications to networks that can support context memory retrieval in the absence of the HPC. The present study aims to identify cortical regions that support the nonHPC context memory. Specifically, the contribution of the perirhinal cortex (PRH) and the anterior cingulate cortex (ACC) were examined because of their established importance to context memory. The findings show that context memories established through distributed reinstatement survive damage limited only to the HPC, PRH, or ACC. Combined lesions of the HPC and PRH, as well as the HPC and ACC, caused retrograde amnesia, suggesting that network modifications in the PRH and ACC enable context fear memories to become resistant to HPC damage.

We test the hypothesis that the stability and precision of context and visual discrimination memories depend on interactions between the hippocampus (HPC) and other memory storage networks. In four experiments we tested the properties of memories acquired in the absence of the HPC. Long–Evans male rats were exclusively used in all experiments. Experiment 1 evaluated acquisition and retention of context fear memories in rats with prior partial or complete HPC damage. Confirming an earlier report (Zelikowsky et al., 2012) a very small but statistically reliable slowing in a single session of context fear conditioning was found after HPC damage. In contrast, retention of context fear memory was normal after HPC damage up to 30 d after learning. In experiment 2, we found that discrimination between a context paired with foot shocks and a different context never paired with foot shock was retained normally for 15 d. In experiment 3, we replicated the finding of intact context discrimination for at least 15 d in rats who display a significant impairment in acquisition of place learning in the Morris water task (MWT). In final experiment using an appetitive object discrimination task, we showed normal retention of the discrimination for at least 30 d after training in rats with complete HPC damage. These finding score against the idea that non HPC memory storage requires a period of interaction with HPC to establish a stable, precise memory.SIGNIFICANCE STATEMENTContrary to expectations from systems memory consolidation, we find that in the absence of a functional hippocampus (HPC) context and visual memories are formed rapidly and exhibit normal persistence and precision. The findings suggest that the HPC is not obligatory for these features of long-term memories.

Education, occupation, and an active lifestyle, comprising enhanced social, physical, and mental components are associated with improved cognitive functions in aged people and may prevent/ or delay the progression of various neurodegenerative diseases including Alzheimer’s disease (AD). To investigate this protective effect, APPNL-G-F/NL-G-F mice at 3 months of age were exposed to repeated, single- or multi-domain cognitive training. Cognitive training was given at the age of 3, 6, 9 & 12 months of age. Single-domain cognitive training was limited to a spatial navigation task. Multi-domain cognitive training consisted of a spatial navigation task, object recognition, and fear conditioning. At the age of 12 months, behavioral tests were completed for cognitive training groups and control group. After completion of behavioral testing, mice were sacrificed, and their brains were assessed for pathology. AppNL-G-F mice given multi-domain cognitive training compared to APPNL-G-F control group showed an improvement in cognitive functions, reductions in amyloid load and microgliosis, and a preservation of cholinergic function. There were mild reductions in microglosis in the brain of APPNL-G-F mice with singledomain cognitive training. These findings provide causal evidence for the potential of certain forms of cognitive training to mitigate the cognitive deficits in Alzheimer disease.### Competing Interest StatementThe authors have declared no competing interest.