The behavior of animals is often complex and requires them to interact with their surroundings. Within the brain, there are specialized neural systems in place to create and store representations of space. In order to effectively utilize these spatial mappings, there must be coordination between sub-cortical systems, which are responsible for allocentric spatial processing, and cortical regions, which handle sensory processing in egocentric coordinates. The retrosplenial cortex is a candidate for the role in facilitating the transformation between different coordinates, as it is anatomically located between the hippocampus and sensory cortical areas and exhibits both egocentric and allocentric spatial responses. The first experiment explored the firing properties of neurons in the retrosplenial cortex in response to boundaries defined in egocentric coordinates. Based on previous research conducted in our lab, which showed that cells in the striatum respond to such boundaries, rats were implanted with electrodes to record the activity of neurons in the retrosplenial cortex while they roamed freely in an open field. The response properties of these neurons were analyzed as the arena was expanded, its shape altered, and boundaries were added and removed. A subgroup of neurons, referred to as Egocentric Boundary Cells, showed increased or decreased firing when the rat was at a specific distance and direction from any of the arena's boundaries. These cells showed no preference for any particular boundary, and their firing was not biased by the animal's angular velocity or turning behavior, suggesting that they respond to boundaries in a general manner, regardless of the features of the boundary or the animal's behavior. Building on experiment one, the second experiment examined the behavior of retrosplenial neurons in rats during open field exploration when alterations were made to the environment. In three separate sessions, a subset of neurons recorded showed either an increase or decrease in their mean firing rate throughout the entire session in which the environment was altered, then returned to prior levels when the environment was returned to a familiar configuration. These alterations included the introduction of an object, rotation of boundaries, expansion of boundaries, changes in the arena's geometry, and removal of boundaries. Similar proportions of neurons exhibited increases or decreases in firing rate for all experimental manipulations. Furthermore, the majority of retrosplenial neurons showed strong speed sensitivity. Some neurons showed an increase in firing rate as speed increased, others showed a decrease in firing rate as speed increased, and some had a specific speed at which their firing rate was the highest. These results support the idea that the RSC is involved in contextual and memory processing, scene processing, as well as transmitting information about self-movement to downstream regions. The third experiment analyzed the different poses of rats as they moved through open field arenas, using simultaneous high-resolution thermal, depth, and RGB cameras. This was organized into a new open-source dataset called Rodent3D. The three-dimensional posture of the animal was reconstructed from the two-dimensional videos using a model called OptiPose. We investigated aspects of the environment where the animals spent the most time looking, such as boundaries and objects, and the frequency and duration of behaviors such as rearing and changes in heading. Finally, we discuss the significance of our model and the potential uses of our unique dataset for the fields of neuroscience and computer vision, as well as future research plans. These experiments demonstrated that the retrosplenial cortex is a vital region for spatial processing, particularly emphasizing egocentric responses. We show that aspects of the environment, such as boundaries, the presence of objects, and changes to the local features induce multiple changes in spatial firing properties of neurons. We also provide a novel open-source model and dataset that provides an innovative tool for more rigorous behavioral analysis that can be used in many disciplines. Altogether, these results suggest that the retrosplenial cortex plays a crucial role as a hub for egocentric spatial processing and coordinating of spatial reference frames to support spatial memory and navigation.

One of the earliest symptoms of Alzheimer’s disease (AD) and age-related cognitive decline is topographical disorientation or impairment to spatial navigation. Furthermore, aging and AD are associated with cortical gray-matter thinning, particularly in the medial temporal and posterior cingulate regions, which have been associated with spatial navigation. Aerobic exercise has been well-established as a beneficial intervention to curtail the neurodegenerative effects of aging. This study aims to explore the relationship between cardiorespiratory fitness (CRF), and two markers of AD and cognitive aging, virtual navigation ability and cortical thickness of the entorhinal, parahippocampal and retrosplenial regions. Cross-sectional data utilized in this study was collected from 23 healthy older adults (60-80 years). Measures included in our analyses consisted of estimated VO2max, T1-weighted structural MR images, and behavioral performance on a virtual navigation task, measured as numbers of objects located during recall. Cortical thickness of the regions of interest (ROIs) was determined by processing T1-weighted MR images in FreeSurfer. We hypothesized that greater CRF would correlate with improved virtual navigation performance and greater cortical thickness of ROIs. Our analyses did not reveal statistically significant relationships between CRF and navigation performance or CRF and cortical thickness. However, Pearson’s correlations found right retrosplenial cortical (RSC) thickness and navigation performance to be significantly related. Multiple regression models of right RSC thickness and navigation performance were performed controlling for age, sex, education and task version. These analyses revealed that greater right RSC thickness predicted navigation performance. Additionally, this model showed that older age predicts decline in navigation performance. Our findings did not survive multiple comparisons correction; nonetheless, the results provide promising insight to the relationship between cortical thickness and navigation performance in healthy aging. Further cross-sectional and longitudinal investigations with a larger sample size are required to assess the impact of CRF and exercise on cortical thickness and navigation abilities in healthy aging. Understanding these relationships would contribute to the expansive body of literature that has linked CRF and exercise to neuroprotective mechanisms in the aging brain.

Path integration is a navigational process that humans and animals use to track changes in their position and orientation. Animal and computational studies suggest that a spatially-tuned navigation system supports path integration, yet this system is not well understood in humans. Here, the prediction was tested that path integration mechanisms and goal-directed navigation in humans would recruit the same key brain regions within the parietal cortex and medial temporal lobes as predicted by animal and computational models. The three experiments described in this dissertation used behavioral and functional magnetic resonance imaging methods in 131 adults (18-35 years) to examine behavioral and brain correlates of navigation. In a landmark-free environment, path integration mechanisms are utilized to update position and orientation to a goal. Experiment 1 examined neural correlates of these mechanisms in the human brain. The results demonstrated that successful first and third person perspective navigation recruited the anterior hippocampus. The posterior hippocampus was found to track distance and temporal proximity to a goal location. The retrosplenial and posterior parietal cortices were additionally recruited for successful goal-directed navigation. In a landmark-rich environment, humans utilize route-based strategies to triangulate between their position, landmarks, and navigational goal. Experiment 2 contrasted path integration and landmark-based strategies by adding a solitary landmark to a sparse environment. The results demonstrated that successful navigation with and without an orienting landmark recruited the anterior hippocampus. Activity in the bilateral posterior hippocampus was modulated by larger triangulation between current position, landmark, and goal location during first person perspective navigation. The caudate nucleus was additionally recruited for landmark-based navigation. Experiment 3 used functional connectivity methods coupled with two fMRI tasks to determine whether areas responsive to optic flow, specifically V3A, V6, and the human motion complex (hMT+), are functionally connected to brain regions recruited during first person perspective navigation. The results demonstrated a functional relationship between optic flow areas and navigationally responsive regions, including the hippocampus, retrosplenial, posterior parietal, and medial prefrontal cortices. These studies demonstrate that goal-directed navigation is reliant upon a navigational system supported by hippocampal position computations and orientation calculations from the retrosplenial and posterior parietal cortices.

The anterior thalamic nuclei (ATN) are a critical part of an extended hippocampal system that supports key elements of episodic memory. Damage or disconnection of the ATN is a component of clinical conditions associated with severe anterograde amnesisa such as the Korsakoff’s syndrome, thalamic stroke, and neurodegenerative disorders. Previous studies have demonstrated that the ATN and hippocampus are often interdependent, and that ATN damage can result in ‘covert pathology’ in ostensibly healthy distal regions of the extended hippocampal system. Adult male rats with neurotoxic bilateral ATN lesions or sham surgery were post-operatively housed in an enriched environment or standard housing after a lesion-induced spatial working memory deficit had been established. These rats were retested on cross-maze and then trained in radial-arm maze spatial memory tasks. Other enriched rats received pseudo-training only after the enrichment period. The detailed neuromorphology of neurons was subsequently examined in the hippocampal CA1. Soma characteristics were also examined in the retrosplenial granular b cortex and the prelimbic cortex. In Experiment 1, ATN lesions produced clear deficits in both the cross-maze and radial-arm maze tasks and reduced hippocampal CA1 dendritic complexity, length, and spine density, while increasing the average diameter of the dendrites. Post-operative enrichment reversed the ATN lesion-induced deficits in the cross-maze and radial-arm maze, and returned CA1 basal and apical spine density to a level comparable to that of sham standard housed trained rats. The sham enriched rats exhibited improved radial-arm maze performance and increased CA1 branching complexity and spine density in both basal and apical arbors compared to sham standard housed rats. The neuromorphological changes observed in the enriched ATN and sham rats may be in part responsible for the spatial working memory improvements observed. Experiment 2 provided support for this contention by demonstrating that the CA1 spine changes were explicitly relevant to spatial learning and memory, because trained enriched sham and ATN rats had increased spines, particularly in the basal tree when compared to closely comparable pseudo-trained enriched rats. Interestingly, spatial memory training increased the numbers of both thin and mushroom spines, whereas enrichment was only associated with an increase in thin spines. In Experiment 3, ATN lesions increased cell body size in layer II of the retrosplenial granular b cortex, whereas enrichment decreased cell body size in layer V of this region. Neither ATN lesions nor enrichment had any effect on cell body morphology in the prelimbic cortex. The current research provides some of the strongest evidence to date of ATN and hippocampal interdependence within the extended hippocampal system, and provides the first evidence of neuromorphological correlates of recovery after ATN lesions.

Encoding and consolidation of memories are two processes within the hippocampus, and connected cortical networks, that recruit different circuit level dynamics to effectively process and pass information from brain region to brain region. In the hippocampal CA1 pyramidal layer local field potential (LFP), these processes take the form of theta and sharp wave ripples (SPW-Rs) for encoding and consolidation, respectively. As an animal runs through an environment, neurons become active at specific locations in the environment (place cells) increasing their firing rate, functionally representing these specific locations. These firing rate increases are organized within the local theta oscillations and sequential activation of many place cells creates a map of the environment. Once the animal stops moving and begins consummatory behaviors, such as eating, drinking, or grooming, theta activity diminishes, and large irregular activity (LIA) begins to dominate the LFP. Spontaneously, with the LIA, the place cells active during the experience are replayed during SPW-Rs in the same spatial order they were encountered in the environment. Both theta and SPW-R oscillations and their associated neuronal firing are necessary for effective place recognition as well as learning and memory. As such, interruption or termination of SPW-R events results in decreased learning performance over days. During exploration, the associated theta and sequential place cell activity is thought to encode the experience. During quiet restfulness or slow wave sleep (SWS), SPW-R events, that replay experience specific place sequences, are thought to be the signal by which systems consolidation progresses and the hippocampus guides cortical synaptic reorganization. The granular retrosplenial cortex (gRSC) is an associational area that exhibits high frequency oscillations (HFOs) during both hippocampal theta and SPW-Rs, and is potentially a period when the gRSC interprets incoming content from the hippocampus during encoding and systems consolidation. However, the precise laminar organization of synaptic currents supporting HFOs, whether the local gRSC circuitry can support HFOs without patterned input, and the precise coupling of hippocmapla oscillations to gRSC HFOs across brain states remains unknown. We aimed to answer these questions using in vivo, awake electrophysiological recordings in head-fixed mice that were trained to run for water rewards in a 1D virtual environment. We show that gRSC synaptic currents supporting HFOs, across all awake brain states, are exclusively localized to layer 2/3 (L2/3), even when events are detected within layer 5 (L5). Using focal optogenetics, both L2/3 and L5 can generate induced HFOs given a strong enough broad stimulation. Spontaneous gRSC HFOs occurring outside of SPW-Rs are highly comodulated with medial entorhinal cortex (MEC) generated high gamma in hippocampal stratum lacunosum moleculare. gRSC HFOs may serve a necessary role in communication between the hippocampus during SPW-Rs states and between the hippocampus, gRSC, and MEC during theta states to support memory consolidation and memory encoding, respectively. / Doctor of Philosophy / As an animal moves through an environment, individual neurons in the hippocampus, known as place cells, increase and decrease their firing rate as the animal enters and exits specific locations in the environment. Within an environment, multiple neurons become active in different locations, this cooperation of spiking in various locations creates a place map of the environment. Now let's say when the animal moved from one corner of the environment to another, place cells 'A', 'C', 'B', 'E', and 'D' became active in that order. This means, at any given point in the environment, the animal is standing in a venn-diagram-esque overlap of place fields, or locations individual place cells represent. A key question that entranced researchers for many years was how do these neurons know when to be active to not impinge on their neighbor's locations? The answer to this question rested with population electrical activity, known as the local field potential (LFP), that place cell activity is paced to. During active navigation through an environment, place cells activity is coupled to the phase of a slow ~8 hertz (Hz) theta oscillation. Within one theta cycle, or peak to peak, multiple place cells are active, representing the venn diagram of location the animal is in. Importantly, this theta activity and encoding of place cell activity is largely seen during active running or rapid eye movement (REM) sleep. During slow wave sleep (SWS), after an animal has experienced a specific environment and has created a place map, place cells are reactivated in the same order the animal experienced them in. From our previous example, the content of this reactivation would be the place cells 'A', 'C', 'B', 'E', and 'D' which all would be reactivated in that same order. These reactivations or replays occur during highly synchronous and fast LFP oscillations known as sharp wave-ripples (SPW-Rs). SPW-Rs are thought to be a key LFP event that drives memory consolidation and the eventual conversion of short-term memory into long-term memory. However, for consolidation to occur, connected cortical regions need to be able to receive and interpret the information within SPW-Rs. The granular retrosplenial cortex (gRSC) is one proposed region that serves this role. During SPW-Rs the superficial gRSC has been shown to exhibit high frequency oscillations (HFOs), which potentially serve the purpose for interpreting SPW-R content. However, HFOs have been reported during hippocampal theta, suggesting HFOs serve multiple purposes in interregional communication across different states. In this study, we found that naturally occurring gRSC HFOs occur exclusively in layer 2/3 across all awake brain states. Using focal optogenetic excitation we were able to evoke HFOs in both layer 2/3 and 5. Spontaneous gRSC HFOs occurring without SPW-Rs were highly comodulated with medial entorhinal cortex (MEC) generated high gamma in hippocampal stratum lacunosum moleculare. gRSC HFOs may serve a general role in supporting hippocampo-cortical dialogue during SPW-R and theta brain states to support memory consolidation and encoding, respectively.

<p><a></a><a>The visual system is capable of interpreting immense sensory complexity, allowing us to quickly identify behaviorally relevant stimuli in the environment. It performs this task with a hierarchical organization that works to detect, relay, and integrate visual stimulus features into an interpretable form. To understand the complexities of this system, visual neuroscientists have benefited from the many advantages of using mice as visual models. Despite their poor visual acuity, these animals possess surprisingly complex visual systems, and have been instrumental in understanding how visual features are processed in the primary visual cortex (V1). However, a growing body of literature has shown that primary sensory areas like V1 are capable of more than basic feature detection, but can express neural activity patterns related to learning, memory, categorization, and prediction. </a></p> <p>Visual experience fundamentally changes the encoding and perception of visual stimuli at many scales, and allows us to become familiar with environmental cues. However, the neural processes that govern visual familiarity are poorly understood. By exposing awake mice to repetitively presented visual stimuli over several days, we observed the emergence of low frequency oscillations in the primary visual cortex (V1). The oscillations emerged in population level responses known as visually evoked potentials (VEPs), as well as single-unit responses, and were not observed before the perceptual experience had occurred. They were also not evoked by novel visual stimuli, suggesting that they represent a new form of visual familiarity in the form of low frequency oscillations. The oscillations also required the muscarinic acetylcholine receptors (mAChRs) for their induction and expression, highlighting the importance of the cholinergic system in this learning and memory-based phenomenon. Ongoing visually evoked oscillations were also shown to increase the VEP amplitude of incoming visual stimuli if the stimuli were presented at the high excitability phase of the oscillations, demonstrating how neural activity with unique temporal dynamics can be used to influence visual processing.</p> <p>Given the necessity of perceptual experience for the strong expression of these oscillations and their dependence on the cholinergic system, it was clear we had discovered a phenomenon grounded in visual learning or memory. To further validate this, we characterized this response in a mouse model of Fragile X syndrome (FX), the most common inherited form of autism and a condition with known visual perceptual learning deficits. Using a multifaceted experimental approach, a number of neurophysiological differences were found in the oscillations displayed in FX mice. Extracellular recordings revealed shorter durations and lower power oscillatory activity in FX mice. Furthermore, we found that the frequency of peak oscillatory activity was significantly decreased in FX mice, demonstrating a unique temporal neural impairment not previously reported in FX. In collaboration with Dr. Christopher J. Quinn at Purdue, we performed functional connectivity analysis on the extracellularly recorded spikes from WT and FX mice. This analysis revealed significant impairments in functional connections from multiple layers in FX mice after the perceptual experience; some of which were validated by another graduate student (Qiuyu Wu) using Channelrhodopsin-2 assisted circuit mapping (CRACM). Together, these results shed new light on how visual stimulus familiarity is differentially encoded in FX via persistent oscillations, and allowed us to identify impairments in cross layer connectivity that may underlie these differences. </p> <p>Finally, we asked whether these oscillations are observable in other brain areas or are intrinsic to V1. Furthermore, we sought to determine if the oscillating unit populations in V1 possess uniform firing dynamics, or contribute differentially to the population level response. By performing paired recordings, we did not find prominent oscillatory activity in two visual thalamic nuclei (dLGN and LP) or a nonvisual area (RSC) connected to V1, suggesting the oscillations may not propagate with similar dynamics via cortico-thalamic connections or retrosplenial connections, <a>but may either be uniquely distributed across the visual hierarchy or predominantly</a> restricted to V1. Using K-means clustering on a large population of oscillating units in V1, we found unique temporal profiles of visually evoked responses, demonstrating distinct contributions of different unit sub-populations to the oscillation response dynamics.</p>