Existing knowledge identifies the prefrontal cortex (PFC) as the modulatory area for pain. Previous neuroimaging studies suggest the existence of the cortico-cortical pathway, an alternative pain modulatory pathway distinct from the descending modulatory pathway of pain. However, little is known of the extent, mechanism and underlying substrate of the modulation. The objective of this study is therefore to explore the role of the PFC in pain modulation. To examine the extent of PFC involvement in pain, meta-analyses of imaging studies in healthy volunteers and patients with chronic pain were performed. Using Gaussian-process regression (GPR) analysis, brain maps were produced from foci of activation as reported in the studies. Since structure dictates function, our next study was to performprobabilistic tractography on diffusion-weighted brain images to ascertain the connection probability of lateral PFC subdivisions and pain-related brain regions as well as intrinsic PFC connections. Two behavioural studies were conducted to investigate cognitive modulation of pain. The first was a study to assess the subjective and physiological correlates of cognitive stress, as previously used in stress-induced analgesia studies. The second was to investigate the involvement of the endogenous opioid system inthe cognitive modulation of pain through effortful reappraisal and contextual modulation. Meta-analyses in healthy volunteers and chronic pain patients revealed activation mainly in the lateral aspect of the PFC due to pain. Distinct pattern of activation was demonstrated in patients with significant ventrolateral PFC (VLPFC) activation across subtypes of chronic pain. Probabilistic tractography further illustrate the functional significance of lateral PFC subdivisions by demonstrating differential connection probability to pain-related brain regions; dorsolateral PFC (DLPFC) regions displayed higher connection probability with brain regions serving more sensory-discriminative function while VLPFC showed high connection probability with both sensory-discriminative and affective regions. Behavioural study of stress showed that cognitive stress failed to induce significant increases in biomarkers of stress, and was not affected by increased level of difficulty. Lastly, behavioural study on contextual modulation and reappraisal confirmed opioid mediation for contextual modulation while negating its involvement in effortful reappraisal. Findings from this studyillustrate the extent of PFC involvement in pain modulation especially in chronic pain patients and provide further evidence of an alternative pathway distinct from the opioid-mediated descending inhibitory pathway.

Physical exercise has repeatedly been shown to trigger positive effects on brain function including improved learning, memory, and executive functions. In addition, corresponding physiological changes have been observed, such as increased neurotrophic factors, changes in neurotransmitter concentrations, and increased dendritic spines. However, these changes have not been well described outside of the hippocampus, including the medial prefrontal cortex (mPFC), and have not been directly compared at different points of development. Because the prefrontal cortex is one of the last brain areas to fully mature, considering the age at which intervention, such as exercise, takes place is particularly important. Additionally, in human studies the data suggest that exercise has the most profound effects on prefrontal-mediated cognitive functions, while there is considerably less evidence on how exercise affects these functions in animals. The experiments presented here draw upon several well-established methodologies to explore the behavioral and physiological changes due to exercise that take place during adulthood compared to adolescence, as well as the role of mPFC sub regions in instrumental extinction and renewal. To that end, these experiments employ conditioning paradigms using appetitive lever-pressing to assess renewal of extinguished instrumental responding following exercise or pharmacological manipulations. Additionally, because there are multiple reports suggesting that early experiences can affect prefrontal neuronal morphology, dendritic length, complexity, and spine density was examined in young or adult male rats that had access to a locked (no exercise) or unlocked (exercise) running wheel for two weeks. Furthermore, norepinephrine transporter (NET) protein expression in the mPFC was examined by Western blot. Collectively, these experiments suggest that exercise in developing, but not adult rats, reduces the expression of instrumental renewal. The precise role of the mPFC and its sub-regions (i.e., prelimbic (PL) and infralimbic (IL)) in instrumental renewal was examined, providing evidence that the behavioral consequences of physical exercise may be due to modifications not only restricted to the mPFC, but also that exercise may have preferential effects on sub-regions, or change the balance of activation. The finding that when juvenile rats exercised they showed less ABA renewal than non-exercisers, paired with the reduction of ABA renewal when the PL was inactivated (and indeed, almost an identical reduction in the two experiments) points to the deduction that exercise is affecting the PL, perhaps more so than the IL or other mPFC regions.

BACKGROUND: Schizophrenia is a debilitating disorder that has a profound impact on quality of life due to the presence of both cognitive deficits and psychotic symptoms. Despite having significant global economic and social costs and a worldwide prevalence of 1%, schizophrenia is still not well understood. Research has been making strides in uncovering the pathophysiology and the etiology that drive this disease, ranging from genetic abnormalities, disrupted circuitry, changes in microarchitecture, to impaired synaptic connectivity. Evidence suggests that disrupted-in-schizophrenia-1 (DISC1) driven genetic disturbances in fast-spiking parvalbumin (PV) neurons and their surrounding perineuronal nets (PNNs) likely contribute to schizophrenia etiology as they are part of the microcircuits required for working memory, a cognitive function that has been consistently impaired in schizophrenic patients. OBJECTIVE: To identify the neuroanatomical changes in PV neurons and surrounding PNNs in the superficial and deep layers of the prelimbic and infralimbic prefrontal cortex of a rat DISC-1 knockout model. METHODS: 19 DISC1-KO male rats and 15 wildtype rats were treated with saline or MK-801. They were sacrificed between P268-269 and brains were extracted and separated at the corpus callosum. After fixing and preserving, the brains were sliced then stained to visualize parvalbumin and perineuronal nets with immunohistochemistry. Slices were imaged and analyzed for PV, PNN, and PV+PNN counts in the superficial and deep regions of the prelimbic and infralimbic cortices. Averages counts within each group were taken and analyzed via 2-way ANOVAs for each brain region and dependent variable. RESULTS: DISC1-KO rats displayed the following trending changes: decreased PV cells in deep layers of infralimbic and decreased PNNs throughout the prelimbic cortex. MK-801 appears to increase the number of unsheathed PV cells in the superficial layers of prelimbic and infralimbic cortex. It decreased the number of PNNs in the prelimbic of wildtype animals but not in the DISC1-KO cohort. MK-801 moderately increased PV counts in DISC1-KO. CONCLUSIONS: This DISC1-KO model is a promising model of schizophrenia as we see the same directionality of decreases in PV and PNN as post mortem human studies. Furthermore, MK-801 is seen to have an increasing trend effect on PV cells, which should be considered when interpreting findings in future studies that look at these markers.

Thesis advisor: Gorica D. Petrovich / The drive to eat, like most motivated behaviors, is controlled by both intrinsic signals from the body as well as extrinsic signals from the environment. Although these factors often act in concert, in some instances environmental cues can override the body’s homeostatic signals. Prior work investigating the ability of learned cues to promote overeating in the absence of hunger identified a critical forebrain network composed of the amygdala, medial prefrontal cortex (mPFC), and lateral hypothalamus (LHA). We hypothesized that a similar forebrain network may also be critical when learned fear-cues inhibit eating despite hunger. The amygdala, mPFC and LHA are each anatomically and functionally positioned to influence feeding, and evidence suggests they could work together to support the fear-cue’s ability to inhibit feeding by overriding homeostatic hunger signals triggered by food-deprivation. Prior anatomical work identified direct pathways between these three large, heterogeneous regions; however, less is known about the organization of the underlying circuitries, especially between distinct nuclei and/or subdivisions that comprise these structures. Study 1 used a dual retrograde tract tracing design to map the topographical organization of the connections between the amygdala, mPFC, and LHA in detail, and to determine whether amygdalar pathways to the mPFC and to LHA originated from the same or different neurons. We found evidence for multiple, topographically organized, direct pathways from the amygdala to the LHA, and separate pathways from the amygdala to areas of the mPFC that send direct projections to the LHA. Importantly, nearly all amygdalar projections to the mPFC and to the LHA originated from different neurons, suggesting that amygdala and amygdala-mPFC processing influence the LHA independently. Study 2 used immediate early gene induction to map the patterns of functional activation within this amygdala-prefrontal-lateral hypothalamic network during the expression of fear-cue inhibited feeding behavior, and to assess whether these patterns were similar in males and females. We found differential activation across the network, and activation patterns related to the presentation of fear-cues, the presence of food-related cues, and the amount of food consumed were associated within distinct cell groups in the amygdala, mPFC, and LHA. Together, the studies presented in this dissertation provide anatomical and functional maps for future interrogation of the circuitry underlying fear-cue inhibited feeding. / Thesis (PhD) — Boston College, 2015. / Submitted to: Boston College. Graduate School of Arts and Sciences. / Discipline: Psychology.

The hippocampus and prefrontal cortex play distinct roles in the generation and retrieval of episodic memory. The hippocampus is crucial for binding inputs across behavioral timescales, whereas the prefrontal cortex is found to influence retrieval. Spiking of hippocampal principal neurons contains environmental information, including information about the presence of specific objects and their spatial or temporal position relative to environmental and behavioral cues. Neural activity in the prefrontal cortex is found to map behavioral sequences that share commonalities in sensory input, movement, and reward valence. Here I conducted a series of four experiments to test the hypothesis that external inputs from cortex update cell assemblies that are organized within the hippocampus. I propose that cortical inputs coordinate with CA3 to rapidly integrate information at fine timescales. Extracellular tetrode recordings of neurons in the orbitofrontal cortex were performed in rats during a task where object valences were dictated by the spatial context in which they were located. Orbitofrontal ensembles, during object sampling, were found to organize all measured task elements in inverse rank relative to the rank previously observed in the hippocampus, whereby orbitofrontal ensembles displayed greater differentiation for object valence and its contextual identity than spatial position. Using the same task, a follow-up experiment assessed coordination between prefrontal and hippocampal networks by simultaneously recording medial prefrontal and hippocampal activity. The circuit was found to coordinate at theta frequencies, whereby hippocampal theta engaged prefrontal signals during contextual sampling, and the order of engagement reversed during object sampling. Two additional experiments investigated hippocampal temporal representations. First, hippocampal patterns were found to represent conjunctions of time and odor during a head-fixed delayed match-to-sample task. Lastly, I assessed the dependence of hippocampal firing patterns on intrinsic connectivity during the delay period versus active navigation of spatial routes, as rats performed a delayed-alternation T-maze. Stimulation of the ventral hippocampal commissure induced remapping of hippocampal activity during the delay period selectively. Despite temporal reorganization, different hippocampal populations emerged to predict temporal position. These results show hippocampal representations are guided by stable cortical signals, but also, coordination between cortical and intrinsic circuitry stabilizes flexible CA1 temporal representations.

Dramatic changes occur across childhood and adolescence in the activity and connectivity of an amygdala-medial prefrontal cortex circuit critical for emotional learning and regulation. However, little is currently known about how neuroplasticity within the circuit changes during development in the human. Experiences that occur during developmental sensitive periods of increased neuroplasticity have the capacity to sculpt neural function with lifelong consequences for cognition and behavior, though. This dissertation will therefore investigate when and how experience may shape amygdala-medial prefrontal cortex functional circuitry (Aim 1) and what the implications of experience-dependent circuitry development are for emotion regulation behaviors (Aim 2) across childhood, adolescence, and adulthood in three studies. Study 1 (previously published as Gabard-Durnam, Gee et al., 2016) posits and tests the long-term phasic molding hypothesis that tonic amygdala-prefrontal cortex functional connectivity, the functional architecture of the brain, is shaped during development by recurring stimulus-elicited connectivity in the circuitry using prospective examination of these connectivities’ development across childhood and adolescence. Study 1 also tests whether the ability of amygdala-prefrontal cortex stimulus-elicited connectivity to shape the amygdala-prefrontal cortex resting-state functional architecture changes across development, reflecting changing plasticity of the circuitry. Study 2 examines how the timing and duration of an early adverse experience, parental deprivation, interacts with genetically-driven differences in neuroplasticity levels indexed by the Brain-Derived Neurotrophic Factor val66met polymorphism to influence the developmental trajectory of amygdala-prefrontal cortex functional architecture using a population of previously-institutionalized children and adolescents and a never-institutionalized comparison sample. Study 2 further examines how the experience- and plasticity-related changes to the functional architecture influence both concurrent and future internalizing symptomatology across childhood and adolescence. Study 3 builds on the first two developmental studies by explicitly testing whether childhood is a sensitive period for medial prefrontal cortex-mediated regulatory signal learning through a retrospective design in adults. Study 3 additionally assesses the effects of developmental experience on adult emotion regulation behavior and physiology. My findings at the levels of brain circuitry, behavior, physiology, and genetics together delineate a period of increased sensitivity to the environment within prefrontal cortex-amygdala functional circuitry from infancy through childhood, modifiable by genetically-conferred variation in plasticity and the nature of the early environment. Moreover, experiences occurring during the sensitive period have consequences for future emotion regulation behavior both during development and lasting into young adulthood. Together, these findings demonstrate how experience-dependent development has enduring effects on amygdala-prefrontal cortex circuitry function and affective behavior.

A hallmark of intelligence in humans and other animals is the ability to engage in complex behaviors geared towards achieving far-removed goals. Such behavior relies on a set of diverse and sophisticated mental processes that are collectively referred to as cognitive or executive in nature. The prefrontal cortex (PFC) has long been considered the primary neural locus for such processes. From humans down to rodents, damage to the PFC has been shown to impair cognition and executive function. In neuropsychiatric disorders such as schizophrenia, dysfunction of the PFC has been strongly linked to cognitive dysfunction. PFC functioning however, necessarily relies on interactions within and between networks of interconnected neurons. Across species, the PFC has been anatomically defined as the region of cortex reciprocally connected with the mediodorsal thalamus (MD), a definition that suggests PFC functioning cannot be divorced from that of its main thalamic counterpart. Indeed, an increasing number of studies have demonstrated the involvement of MD in cognitive behaviors. Schizophrenia patients performing cognitive tasks also exhibit decreased MD activity, with growing evidence for decreased functional connectivity with the PFC. The studies presented here seek to build on this literature using the mouse as a model organism. Taking advantage of recent tools for temporally- and spatially-restricted manipulations of neural activity we show that a relatively mild and reversible decrease in MD activity is capable of impairing two cognitive behaviors classically shown to be PFC-dependent – behavioral flexibility and working memory. Simultaneously recording MD and PFC activity while mice perform a spatial working memory task, we show task modulations of synchronous MD-PFC activity that are disrupted by a primary decrease in MD activity. Following up on this finding using pathway-specific manipulations of MD-to-PFC and PFC-to-MD neural connections, we provide behavioral and neurophysiological evidence that these circuits serve as distinct neural substrates for working memory maintenance and retrieval. Together, these findings provide causal evidence in support of an association between thalamo-prefrontal dysfunction and cognitive impairment, and may enable the development of more selective therapeutic strategies for cognitive disorders such as schizophrenia.

The ability to take in one’s environment, integrate relevant information, and then act appropriately is an incredibly complex feat that organisms do continuously. Disruption in the ability to think and act clearly, or cognitive dysfunction, is a debilitating aspect of neuropsychiatric diseases like schizophrenia. The prefrontal cortex and the striatum are key brain regions for functional and dysfunctional cognition, but the way that they interact to allow for cognitive processing is poorly understood. To get at these questions, I manipulated and recorded from medial frontal and striatal neurons—frontostriatal ensembles—while rats engaged in interval timing, an elementary cognitive function that engages both areas. I report four main results. First, ramping activity—a gradual, consistent change in neuronal firing rate across time—is observed throughout frontostriatal ensembles. Secondly, medial frontal areas dynamically reflect changing temporal conditions during learning and precede these same changes in striatal areas. Thirdly, interval timing and striatal ramping activity are disrupted when the medial frontal cortex is inactivated. Finally, this behavioral impairment can be reduced by optogenetic stimulation of frontostriatal terminals. My results support the view that striatal neurons integrate medial frontal activity and suggest a possible mechanism—ramping activity—through which neurons might represent the passage of time. These observations elucidate temporal processing in frontostriatal circuits and provide insight into how the medial frontal cortex exerts top-down control of cognitive processing in the striatum. My hope is that these findings will contribute to a clearer understanding of basic cognitive processing and might inform future approaches to treatments that address cognitive dysfunction.

Fear generalization, the generalization of fear to innocuous stimuli, is a characteristic component of pathological anxiety. For example, after returning from war, a person might begin to experience fear in response to the sound of fireworks, a stimulus typically regarded as safe. When excessive, “overgeneralization” serves as a core feature of fear and anxiety-related disorders, such as PTSD. The present collection of studies sought to investigate the neural correlates and temporal dynamics of fear generalization in humans. The first study sought to investigate the causal role of the ventromedial prefrontal cortex (vmPFC) and hippocampus in the generalization of fear. Contrary to hypotheses, individuals with focal damage to the vmPFC (N=8) or hippocampus (N=12) did not demonstrate significantly increased fear generalization relative to individuals with brain damage outside of these regions (N=16) or normal comparison participants (N=20). Potential explanations for this finding are explored. The second study sought to investigate the time course of fear generalization in humans. Participants (N = 107) completed a fear generalization task over the course of two sessions. Results indicate that fear generalization significantly increased as the duration of time between training and testing increased. This suggests that a stimulus may elicit a generalized fear response at one arbitrarily selected time point, but not another. This study establishes a novel paradigm that can be used in future work to investigate changes in the neural correlates of fear generalization over time. Fear generalization is found across an array of anxiety disorders, making it a compelling area of study. The present work highlights the dynamic nature of fear generalization in humans. Further, the present study leads to a number of questions for future research.

Emotion regulation is an ability that humans engage in throughout their lives. Disruption in this ability due to brain injury can have devastating consequences on the ability to function adaptively in complex environments. It has been observed that damage involving certain areas of the prefrontal cortex (PFC), including the ventromedial PFC (VMPFC), can result in long-lasting impairments in real-world emotional and behavioral functioning. However, the specific areas of the PFC that are critical for the ability to regulate emotion have not been identified. The primary aims of this project were to identify areas of the PFC that are important for the regulation of emotion, and to determine the degree to which impairments in emotion regulation may contribute to real-world dysfunction following damage to the PFC. To address these aims, emotional regulation and real-world functioning were examined in a sample of patients with focal PFC lesions. Damage involving the VMPFC appeared to have limited impact on the ability to voluntarily regulate emotion. It was also observed that damage to PFC regions outside the VMPFC was associated with reduced ability to overcome distraction by salient emotional stimuli, compared to VMPFC damage. However, analyses of lesion volume showed that more extensive damage involving the VMPFC was associated with greater emotional distraction, suggesting one form of emotional dysregulation that may result from damage to the VMPFC. In addition, it was found that brain damage in general was associated with impairments in real-world functioning, though PFC damage was not associated with more striking impairments compared to damage outside the PFC. These findings suggest that damage involving certain PFC regions can disrupt the ability to effectively regulate emotion. The results from this project also suggest that laboratory measures of emotion regulation may help in predicting real-world dysfunction following brain damage.