Conversation is a highly interactive and coordinated effort between interactants. For example, interactants often mimic the behaviors and speech of one another and coordinate the timing of behaviors, or interactional synchrony. Despite being affected in certain neurological and psychiatric disorders, the neural mechanisms underlying these processes are not understood. The goal of this study is to understand the role of the ventromedial prefrontal cortex (vmPFC), an area of the brain involved in social and emotional behavior, for interpersonal coordination, including mimicry and interactional synchrony. To test the role of the vmPFC for mimicry, normal comparison (NC), brain damaged comparison (BDC), and participants with vmPFC damage interacted in two sessions with a research assistant (RA) who was performing a target behavior (1st session: nodding, 2nd session: face touching). The amount of time the participants spent nodding or touching their face in each session was recorded. NC and BDC participants tended to mimic the partner and nodded slightly more in the session in which the RA was nodding, and touched their face slightly more in the session in which the RA was touching their face. In contrast, vmPFC patients showed no difference in their behaviors in either session, suggesting that they were not influenced by the partner's behaviors and did not mimic them. In a second experiment, all of the above participant groups had a naturalistic conversation with an unfamiliar interactional partner. The conversational data were analyzed for numerous aspects of interpersonal coordination, including convergence of number of words, words per turn and backchannels, reciprocity of self-disclosures, the use of questions, interactional synchrony, and a time series analysis of response latency and speech rate. The vmPFC participants performed consistently worse than NC participants on convergence of words and words per turn, self-disclosures and asking questions. All brain-damaged participants were impaired on aspects of interactional synchrony, and no conclusive results were found for the time series analysis of response latency and speech rate. This study provides support for the hypothesis that the vmPFC is important for interpersonal coordination as the vmPFC group differed significantly from the NC group on the majority of the analyses. The final goal of this study was to understand the effects of traumatic brain injury (TBI) on interpersonal coordination. TBI patients participated in all of the experiments described above and preliminary results showed that they also seemed to be impaired on the mimicry task, and they performed slightly worse than NC participants on many of the interpersonal coordination analyses of the conversational data. This suggests that TBI also does seem to affect certain aspects of interpersonal coordination.

My thesis research comprises two projects looking into physiological functions of Drosophila ion channels: first, contribution of several T ransient R eceptor P otential (TRP) channels to gravity sensing; second, regulation of metabolic homeostasis by a D egenerin/ E pithelial Na + C hannel (DEG/ENaC). Many animal species sense gravity for spatial orientation. In humans recurrent vertigo and dizziness are often attributable to impairment of gravity sensing in the vestibular organs. However, the molecular bases for gravity sensing and its disruption in vestibular disease remain uncertain. Here I studied gravity sensing in the model organism Drosophila melanogaster, with a combination of genetic, behavioral and electrophysiological methods. My results show that gravity sensing requires Johnston’s organ, a mechanosensory structure located in the antenna that also mediates hearing. Johnston’s organ neurons fire action potentials in a phasic manner in response to body rotations in the gravitational field. Furthermore, gravity sensing and hearing require different TRP channels with distinct anatomical localizations, implying separate neural mechanisms underlying gravity sensing and hearing. These findings set the stage for understanding how TRP channels contribute to the sensory transduction of gravity. Drosophila melanogaster has over 20 genes belonging to the DEG/ENaC family, which are collectively referred to as pickpockets (ppks) . Genetic analyses have implicated ppk genes in salt taste, tracheal liquid clearance, pheromone detection, and developmental timing. These results, together with the conserved presence of DEG/ENaC genes through evolution, suggest that further studies on fly ppk genes may help gain insights to a number of physiological processes. Here I report that the ppk11 gene regulates metabolic homeostasis. A ppk11 enhancer/promoter fragment labels the fat body, the lipid storage organ of Drosophila. ppk11 mutants are lean — they store less triacylglyceride (TAG), possess smaller lipid droplets and are sensitive to starvation compared to wild–type flies. ppk11 mutants also show signs of enhanced insulin sensitivity — they store more glycogen and maintain a lower level of circulating carbohydrate (trehalose). Moreover, the mutants have extended life span, suggesting ppk11–dependent activities of the fat body have systematic and long–term effects on the fly body. Understanding the cellular function of ppk11 may offer new insights into mechanisms that regulate metabolic homeostasis.

Traumatic experiences, such as natural disasters, do not only cause people to suffer from material and financial losses, but they can lead to a maladaptive regulation of the stress response and to the onset of stress-induced psychopathology. Traumatic stress has been shown to alter brain structures and functions involved in the stress response. It has also been linked to the dysregulation of the hypothalamic-pituitary-adrenal axis, an overactive sympathetic nervous system with elevated cortisol and norepinephrine levels. These neurobiological alterations can make some individuals more vulnerable to the development of depression and anxiety disorders. A dose-response relationship between trauma severity and psychopathology has been found in previous research. Research has also revealed that a person’s perceived ability to cope with hardship, coping self-efficacy (CSE), is related to decreased vulnerability or resilience to stress. In the study carried out in the framework of this thesis, associations between the severity of traumatic exposure and CSE with posttraumatic stress disorder, depression, and generalized anxiety disorder were examined in a sample (N = 105) of Nepalese flood victims. Participants (18-90 years old) answered a questionnaire carried out via interview. Results showed that there were no significant correlations between flood related trauma severity and depression or anxiety. However, findings showed that higher CSE was associated with fewer depressive symptoms. Future studies in Nepal should directly investigate this association as well as possible interventions aimed at enhancing CSE and whether such interventions can reduce symptoms of depression.

Previous studies completed with humans have revealed insight into the effects of anxiety on attention tasks such the dot-probe task, but there is little information about such effects on non-human primates. This study aimed to assess whether anxiety or anxious behaviors would impact rhesus macaque performance on a three stimuli paradigm similar to the dot-probe task. Utilizing images of conspecifics (strong threat, mild threat, and neutral), eight monkeys were video recorded completing a task that required them to slide two doors, which held these images, to the side to obtain a treat. We hypothesized that behavioral phenotype (high or low anxiety) would affect attention on this modified dot-probe task. Additionally, we predicted that time spent looking at mildly threatening stimuli would be positively correlated with high levels of anxious behaviors (e.g., scratching, yawning, pacing, self-biting) and cortisol concentrations over a four month period. We also predicted that a higher percentage of the mildly threating stimuli as a first choice would be positively correlated with high levels of anxious behaviors and cortisol concentrations. However, anxious behaviors and cortisol concentrations did not affect performance on this task. Interestingly, a sex difference was found for the mild threat stimuli, with females taking significantly more time to complete the task when presented with the mild stimuli (p = 0.01), and also looking at the mild stimuli longer than males (p = 0.03). These data suggest that males and females interpret ambiguous facial expressions differently, possibly indicating the significance of attention in female dominance hierarchies in macaque social groups.

Maternal behavior is the complex array of caregiving behaviors females display towards offspring. In rats, the transition to motherhood depends on the action of various hormones, especially estradiol near parturition, which primes the maternal circuitry to respond to pups upon first encounter at parturition with appropriate maternal behavior. Although virgin rats avoid pups, new mothers are highly motivated to interact with pups, and their maternal behavior depends on the functional interaction between the medial preoptic area (mPOA) and the ventral tegmental area (VTA). However, a precise mapping of the VTA-projecting mPOA neurons remains to be elucidated. To determine whether pup-responsive neurons in the mPOA project to the VTA, we injected the retrograde tracer Fluorogold (FG) into the VTA of new mother and virgin female rats. Six days later, females were exposed to 3 pups for 5 minutes, and their brains processed to visualize FG and c-Fos immunostaining. In addition, we further characterized the molecular phenotype of these neurons by performing immunohistochemistry against estrogen receptor alpha (Esr1). As expected, the behavior of postpartum and virgin females toward pups was different. Mothers readily approached pups and displayed maternal behavior, whereas virgins avoided interaction with pups. Despite these disparate responses to pups, no differences were found in the number and distribution of mPOAc-Fos→VTA neurons. In addition, in both postpartum and virgin females, a significant proportion of these pup-responsive mPOA→VTA projecting neurons also express Esr1. Further functional interrogation of these c-Fos+/Esr1+ mPOA→VTA neurons in virgins and mothers might elucidate distinct circuit dynamics potentially underlying their behavioral differences towards pups.

Long term cognitive studies in humans and nonhuman primates such as macaques are difficult because of their long lifespan. The common marmoset (Callithrix jacchus) is a non-human primate who shares with humans many features characteristic of primates, including a complex brain and cognitive function. They also have a short lifespan (~10 years) that makes them a great model in studies of cognitive aging. This study focuses on the rate of decline in cognitive function in male and female marmosets based on performance on reversal learning tasks over 2 years of testing. We found that marmosets improved their overall performance from Year 1 to Year 2 due to practice effect, but that females exhibited an impairment in reversal learning compared to males in both years. We also found important individual differences, with some monkeys showing decline in Year 2 compared to Year 1 while most monkeys maintained or improved their performance in reversal learning over the two years. We conclude that (1) cognitive flexibility, as assessed by reversal learning, is impaired in middle-aged female marmosets compared to males, likely due to sex differences in habitual vs. goal-directed behavior, and (2) that reversal learning is a sensitive measure that can capture one year individual changes in cognitive function.

This study examined the relationships between motivation and cognitive performance in male and female common marmosets (Callithrix jacchus). This question was driven by prior data from the Lacreuse lab showing a robust female impairment in reversal learning, as assessed by the number of trials needed to acquire a reversal following a simple discrimination between two stimuli. This thesis tested the hypothesis that the female impairment in reversal learning was mediated by deficits in motivation. Two sets of measures were used to test this hypothesis. I evaluated physical effort via testing on the progressive ratio (PR), a test that requires animals to produce an increasing number of responses to get a reward. Cognitive effort was evaluated through the number of refusals (aborted trials) produced during performance of a reversal. Because estrogen replacement was previously shown to impair reversal learning in ovariectomized females, I also examined whether PR performance was affected by estrogen levels in a subset of female subjects. Contrary to my hypothesis, reversal learning was not significantly associated with cognitive or physical effort in either males or females. Estrogen levels did not significantly affect physical effort, but there was too much variability in a small sample of females for these results to be compelling. We conclude that the sex difference in reversal learning performance is unlikely to be mediated by sex differences in motivation. Instead females may be more likely than males to engage in habitual processes implicating the dorsal striatum, likely through the action of estrogens on this brain region.

The medial amygdala (MeA) is a central node in the interwoven circuits that regulate social behavior based on pheromones. Aromatase-expressing (arom+) neurons in the MeA are key for the establishment and maintenance of sex differences. Here, we characterized the intrinsic electrophysiological properties of arom+ neurons and non-aromatase (arom-) neurons in the MeA of male and female mice. Most electrophysiological properties were similar for arom+ neurons in the MeA between sexes, but the relative refractory period was twice as large in female mice. We also show that the firing pattern and firing frequency is markedly different between arom+ and arom- neurons. The activity of MeA neurons could be modulated by estradiol, which reduced activity in arom+ neurons in males. The differences between arom+ and arom- neurons were observed in both sexes suggesting that aromatase expression delineates a neural population in the MeA with similar and unique electrophysiological properties.

Glia, whose name derives from the original Greek word, meaning “glue,” have long been understood to be cells that play an important functional role in the nutritive and structural support of the central nervous system, yet their full involvement has been historically undervalued. Despite the strong evidence that glial reactions to cellular debris govern the health of the nervous system, the specific properties of damaged axonal debris and the mechanisms by which glia sense them, morphologically adapt to their presence, and initiate phagocytosis for clearance, have remained poorly understood. The work presented in this thesis was aimed at addressing this fundamental gap in our understanding of the role for glia in neurodegenerative processes. I demonstrate that the cellular machinery responsible for the phagocytosis of apoptotic cell corpses is well conserved from worms to mammals. Draper is a key component of the glial response machinery and I am able to show here, for the first time, that it signals through Drosophila Shark, a non-receptor tyrosine kinase similar to mammalian Syk and Zap-70. Shark binds Draper through an immunoreceptor tyrosine-based activation motif (ITAM) in the Draper intracellular domain. I show that Shark activity is essential for Draper-mediated signaling events in vivo, including the recruitment of glial membranes to axons undergoing Wallerian degeneration. I further show that the Src family kinase (SFK) Src42A can markedly increase Draper phosphorylation and is essential for glial phagocytic activity. Therefore I propose that ligand-dependent Draper receptor activation initiates the Src42A-dependent tyrosine phosphorylation of Draper, the association of Shark and the subsequent downstream activation of the Draper pathway. I observed that these Draper-Src42A-Shark interactions are strikingly similar to mammalian immunoreceptor-SFK-Syk signaling events in myeloid and lymphoid cells. Thus, Draper appears to be an ancient immunoreceptor with an extracellular domain tuned to modified-self antigens and an intracellular domain that promotes phagocytosis through an ITAM domain-SFK-Syk-mediated signaling cascade. I have further identified the Drosophila guanine-nucleotide exchange factor (GEF) complex Crk/Mbc/dCed-12, and the small GTPase Rac1 as novel modulators of glial clearance of axonal debris. I am able to demonstrate that Crk/Mbc/dCed-12 and Rac1 function in a non-redundant fashion with the Draper pathway to promote a distinct step in the clearance of axonal debris. Whereas Draper signaling is required early during glial responses, promoting glial activation and extension of glial membranes to degenerating axons, the Crk/Mbc/dCed-12 complex functions at later stages of glial response, promoting the actual phagocytosis of axonal debris. Finally, many interesting mutants have been identified in primary screens for genes active in neurons that are required for axon fragmentation or clearance by glia, and genes potentially active in glia that orchestrate clearance of fragmented axons. The further characterization of these genes will likely unlock the mystery surrounding “eat me” and “find me” cues hypothesized to be released or exposed by neurons undergoing degeneration. Illuminating these important glial pathways could lead to a novel therapeutic approach to brain trauma or other neurodegenerative conditions by providing a druggable means of inducing early attenuation of the glial response to injury down to levels less damaging to the brain. Taken together, my combined work identifies new components of the glial engulfment machinery and shows that glial activation, phagocytosis of axonal debris, and the termination of glial responses to injury are genetically separable events mediated by distinct signaling pathways.

The human neocortex is a highly sophisticated and organized brain structure that is thought to mediate some of the most complex cognitive functions in humans including language and abstract thought. As such, environmental and genetic insults to its normal structure or function can result in devastating neurological conditions including severe epilepsy and intellectual disability. Malformations of cortical development are an increasing collection of disorders that cause neocortical abnormalities due to impaired developmental processes. One recently identified disorder in this class is a thin lissencephaly variant (TLIS) associated with several mutations in the C-terminus death domain of the caspase-2 activation adaptor CRADD (also known as RAIDD). Beyond this, little is known about the mechanism underlying TLIS pathophysiology despite an increasing number of identified individuals suffering from it. In order to better understand this disorder, as well as the normal developmental mechanisms that are impaired in its pathogenesis, I have developed and characterized three murine models by introducing one of a number of different genetic perturbations associated with TLIS. These animal models show behavioral and biochemical abnormalities similar to those seen in human TLIS subjects. Focusing future studies on the developmental processes that underlie differences seen in these mouse models could greatly inform understanding of disease mechanism in humans and assist in the development in therapeutic interventions. My work presented in this dissertation thus effectively establishes a translationally relevant animal model of TLIS.