abstract / Anatomy / Master / Master of Philosophy

Two studies of voluntary movement in Parkinson's disease were carried out. In the first study, both parkinsonians and age-matched controls performed unconstrained prehensile movements in which subjects produced reaching and grasping (hand opening/closing) movements under varying conditions of movement amplitude, speed and object size. The act of prehension requires the intersegmental coordination of limb transport and grasp trajectories. Although parkinsonians were slower overall than controls, patients and controls similarly adjusted the spatial and temporal characteristics of their limb movement and grasp in response to changes in task demands. All groups showed increases in the speed of limb transport, the speed of hand opening and began hand opening proportionately earlier at faster movement speeds. All groups similarly increased hand opening velocity and initiated grasp earlier for smaller amplitude movements. Likewise, grasp was initiated earlier when reaching for wider objects. However, in contrast to controls, the onset time of hand opening during limb transport was delayed in these patients and was found to be more closely coupled with the timing of limb transport than in the controls. Moreover, patients showed greater curvature in their motion paths at the wrist during limb transport suggesting that the timing of joint motion (shoulder and elbow) may be different in these patients as well. Underlying differences between the groups in the temporal sequencing of movement are discussed. / In a second study, parkinson and control subjects performed continuous tracking movements in pursuit of sinusoidal and constant-speed target trajectories varying in frequency and amplitude. This task provided explicit temporal and spatial accuracy constraints by requiring subjects to reproduce the precise trajectory (i.e., velocity profile) of target movement. The results show that patients, similar to controls, were capable of modifying peak movement velocity while varying their times to reversal (i.e., movement durations) in response to changes in the movement time requirements of target motion. Indeed, both patients and controls were shown to alter the timing of movement deceleration in order to maintain their movement durations within the temporal limits of target movement. In contrast to controls, patients show progressive reduction in endpoint accuracy (undershooting the target) and, hence, reduced movement amplitudes, over the course of the trial. However, when endpoint accuracy requirements were reduced, by providing mechanical constraints on movement amplitude, patients were able to increase movement amplitudes while satisfying the temporal requirements of the task. These results are interpreted in terms of tradeoffs in performance between competing spatial and temporal demands of pursuit tracking. The significance of movement accuracy constraints on motor function in parkinsonian performance is discussed.

Parkinson’s disease (PD) is a multi-organ disorder. It is diagnosed from motor impairments that arise from neurodegeneration in the midbrain. However, the disease begins decades earlier in the gut prior to involvement of the brain. PD is characterized by persistent inflammation, both in the brain and in the periphery in addition to neurodegeneration. Here, I investigate the role of the adaptive immune system in disease pathogenesis and as a driver of prodromal symptoms of PD in both humans and mice. In Chapter 1, I introduce Parkinson’s disease, its pathological hallmarks and the progression of the symptoms, and discuss genetic and environmental influences. Then, I elaborate on the inflammatory phenotypes observed in the disease and recent work describing the role of inflammation in animal models for PD. In Chapter 2, I examine the autoimmune features of Parkinson’s disease from analysis of patients’ blood. I found that approximately 40% of PD patients possess aspects of autoimmunity against α-synuclein. By screening peripheral blood mononuclear cells of patients and healthy controls for potential neoantigens derived from α-synuclein protein, I identified two antigenic regions of the protein that elicit an immune response. The immune responses to a specific α-synuclein neo-antigens were linked to unique HLAs that are over-represented in our PD cohort and are associated with PD in genome wide association studies (GWAS). In Chapters 3 and 4, I describe the effects of recapitulating α-synuclein autoimmunity in a humanized mouse strain expressing the HLA allele risk for PD. In Chapter 3, I show that the humoral and cellular immunity is mounted against α-synuclein in the humanized mice, similar to what is observed in PD patients; however, there is no inflammation or immune response toward the brain. In Chapter 4, I show how the autoimmune response to α-synuclein induces inflammation and neurodegeneration in the gut leading to constipation in mice, recapitulating the prodromal aspects of the human disease. Finally, in Chapter 5, I discuss the implications of these findings for α-synuclein autoimmunity in the periphery, gut and brain in Parkinson’s disease. I also elaborate on the implications of these findings for potential future diagnostic screening and treatments for Parkinson’s disease.

No description available.

Respiratory-swallow coordination (RSC) is critical for safe and efficient swallowing. In healthy adults, RSC is most frequently characterized by an exhale-swallow-exhale pattern initiated within the mid-lung volume range with a respiratory pause of approximately one second. This combination in RSC behaviors is thought to be most optimal for swallowing-related bolus clearance and airway protection. Deviations from these RSC behaviors are observed at disproportionately higher rates in people with Parkinson’s disease (PD) when compared to non-dysphagic, healthy adults. However, little is known about which variables influence RSC in PD, if the RSC behaviors that are most optimal for swallowing in healthy adults are also most optimal for swallowing in PD, and if respiratory-swallow training can be used to successfully rehabilitate suboptimal RSC, swallowing safety, and swallowing efficiency in PD. This dissertation document includes a series of four studies that address these important clinical research questions. Chapter 1 will begin by reviewing the current body of literature as it relates to dysphagia in PD, RSC in healthy adults and PD, respiratory-swallow training as a skill-based treatment for dysphagia rehabilitation, and motor learning considerations for respiratory-swallow skill training in PD. Chapter 2 will be used to examine the relationships among RSC with patient- and swallowing-specific factors in PD. Chapter 2 will also be used to assess the influence of RSC behaviors on measures of swallowing safety (penetration-aspiration) and swallowing efficiency (pharyngeal residue) in PD. Chapter 3 will then evaluate the effects of verbal cueing on RSC in PD as a method of determining if RSC is stimulable for rehabilitative change. Chapter 4 will explore the effects of respiratory-swallow training on swallowing safety and efficiency rehabilitation in a person with mid-stage PD and severe dysphagia within the context of a single-subject experimental design. Chapter 5 will then examine the effects of respiratory-swallow training on dysphagia and RSC rehabilitation within the context of a cohort study. Chapter 5 will also be used to compare the effects of constant versus variable practice in order to explore how principles of motor learning can be used to enhance respiratory-swallow training outcomes. This document will then conclude by synthesizing the results from Chapters 2-5 and by discussing directions for future research.

Parkinson’s disease (PD) is characterized by the degeneration of midbrain dopaminergic neurons. Genetic studies have revealed causative and risk loci associated with a proportion of PD cases, such as PRKN/PARK2, encoding parkin and when mutated causes a rare familial form of autosomal recessive PD. Cell-based studies have linked parkin to mitochondrial turnover by autophagy, but to date, manipulating this gene in rodents has not robustly recapitulated core features of PD. Reconciling these results is essential to determine parkin’s role in mitochondrial biology, brain physiology, and PD pathogenesis. Here, we find that global, inducible deletion of Prkn/Park2 (parkin iKO) in the adult mouse leads to age-dependent motor impairments that are responsive to levodopa treatment. We report that these behavioral defects are associated with progressive pathology in dopaminergic neurons, regional gliosis and lipid oxidation changes, culminating in the selective degeneration of nigrostriatal dopaminergic neurons. We also present a new, in vivo mitophagy reporter system to investigate the relationship of parkin’s described roles in mitochondrial homeostasis to the observed phenotypes. These results give critical insight into parkin’s contribution to dopaminergic neuron stability in the mammalian brain, and provide two distinct and novel organismal tools to investigate mitochondrial homeostasis and PD pathogenesis.

Myeloid cells of the innate immune system have been strongly implicated in the pathogenesis of neurodegenerative diseases, including Alzheimer’s disease (AD) and Parkinson’s disease (PD). Similarly, several lines of evidence call on the adaptive immune system as a critical driver of disease, particularly in PD. The immune dynamics in both of these diseases are complex, and span across not only the innate and adaptive immune systems, but also across the periphery local action in the central nervous system (CNS). This thesis aims to address critical gaps in our knowledge regarding molecular and functional alterations of immune cells in AD and PD. We apply tools including single nucleus RNA – and ATAC – sequencing as well as protein – level and functional studies to advance our understanding of molecular pathways involved in the innate and adaptive immune dysfunction in these diseases, including both immune cells in the CNS as well as in the periphery. Chapter 1 provides an overview of the evidence implicating myeloid cell dysfunction in AD and PD, including microglia as well as peripheral myeloid cells such as monocytes. It also describes the features of immune dysregulation in both diseases, and evidence implicating the adaptive immune system in PD. Chapter 2 aims to address our currently limited understanding of microglial molecular phenotypes and diversity in PD, by characterizing microglial transcriptomic and chromatin signatures in disease. We demonstrate microglial subpopulation-specific effects, including the focal depletion of a microglial population in the substantia nigra in PD, which open novel avenues for targeted neuroimmune interventions in PD. Chapter 3 aims to identify interactions regulating the infiltration and retention of peripheral immune cells into the CNS in PD; a process which is implicated in the progression of this disease, but the mechanisms of which are not fully understood. We characterized transcriptomic signatures of infiltrating lymphocytes and blood brain barrier cells, and found increased T cell infiltration in PD as well as fibroblast and endothelial populations associated with disease. We further identified transcriptional shifts suggestive of a proinflammatory and profibrotic milieu in disease, in which chemokines and extracellular matrix elements produced by fibroblasts may influence T cell trafficking and retention in the substantia nigra in PD. Chapter 4 aims to address the gap in our knowledge of how myeloid dysfunction in the periphery contributes to AD. While genetics implicate all myeloid cells in AD and PD, contributions of peripheral myeloid cells, such as monocytes, have been largely overlooked in place of microglia, which are resident in the CNS. We evaluate the convergence of the AD genetic risk loci on functional outcomes in monocytes, in the context of Aβ as an immune stimulus. We identified functional convergence of the CD33 and SPI1 AD risk variants in the context of Aβ stress, including reduced phagocytosis and loss of surface TREM2 expression, demonstrating an interaction between genetics and environment to reduce myeloid cell fitness. Finally, Chapter 5 concludes with a summary of key findings from this work, and discusses future directions for modulating innate and adaptive immune populations, both in the CNS and in the periphery, as therapeutic approaches for these neurodegenerative diseases.

Adult neurogenesis is affected in neurodegenerative diseases and also represents an important therapeutic target. The goal of this dissertation research was to test the hypothesis that regeneration of neurons and glia in the adult brain can be manipulated by neurotrophic drugs in the context of two mouse models of neurodegenerative disease : Parkinson's disease and Huntington's disease.... These findings have implications for the pathophysiology of Huntington's disease and neurodegeneration in general. Specific alterations to the SVZ neurogenic niche parallel some of the pre-motor symptoms of Parkinson's disease and Huntington's disease. This dissertation research contributes to the growing body of literature concerning the pharmacological modulation of SVZ-derived neurogenesis designed to attenuate the progressive loss of neurons in neurodegenerative diseases and perhaps delay the onset of symptoms. / by Mark Harvey McCollum. / Vita. / Thesis (Ph.D.)--Florida Atlantic University, 2012. / Includes bibliography. / Mode of access: World Wide Web. / System requirements: Adobe Reader.

T cells infiltrate the degenerating brain and influence central nervous system (CNS) inflammation and neuronal health. In mice, the choroid plexus and the meninges have been implicated in regulating T cell entry and egress from the CNS, respectively. Further, antigen presenting cells in the mouse meninges present CNS-derived antigens to T cells and may represent a method for the peripheral immune system to sense and respond to CNS immune triggers. However, whether these processes occur in the human choroid plexus and meninges has not been comprehensively studied. Further, the antigens towards which T cells in the degenerating human brain and its borders respond remain unknown. Therefore, I implemented a multi-omics approach using fresh postmortem tissue from patients diagnosed with amyotrophic lateral sclerosis (ALS), Alzheimer’s disease (AD), Parkinson’s disease (PD), and non-neurodegenerative controls to identify not only the T cell-associated changes that occur in the degenerating human CNS and surrounding tissues but also identified a library of putative antigen targets for disease-associated T cell populations. Specifically, using single cell RNA and TCR sequencing information from paired postmortem choroid plexus, leptomeninges, and brain I lineage traced T cells using their TCR information and found that T cell access to leptomeninges and brain is likely limited and controlled by anti-inflammatory macrophage activity at the blood/CSF barrier (BCSFB). Once past the BCSFB, I present evidence that T cells access the CNS where they interact with MHC expressed by microglia. T cells also accumulate in the leptomeninges where they become tissue resident memory T cells. These tissue resident memory T cells likely serve as a reservoir for a rapid antigen-driven immune response to future CNS inflammatory insults. Finally, by performing immunopeptidomics to identify peptides presented by MHC in the same patients’ CNS and border tissues, I identified a library of putative antigenic triggers that may drive high levels of T cell clonal expansion in the brain and surrounding tissues. Altogether, this thesis serves as a resource for understanding the trajectory of T cells as they travel into the degenerating human brain and as a foundation for the development of antigen-specific precision medicines to treat neurodegeneration.