The role of intracellular calcium perturbations in muscle damage and dysfunction in mouse models of muscular dystrophy

Duchenne muscular dystrophy (DMD) is a neuromuscular disease caused by mutations in the dystrophin gene. DMD is clinically characterized by severe, progressive and irreversible loss of muscle function, in which most patients lose the ability to walk by their early teens and die by their early 20’s. Impaired intracellular calcium (Ca2+) regulation and activation of cell degradation pathways have been proposed as key contributors to DMD disease progression. This dissertation research consists of three studies investigating the role of intracellular Ca2+ in skeletal muscle dysfunction in different mouse models of DMD. Study one evaluated the role of Ca2+-activated enzymes (proteases) that activate protein degradation in excitation-contraction (E-C) coupling failure following repeated contractions in mdx and dystrophin-utrophin null (mdx/utr-/-) mice. Single muscle fibers from mdx/utr-/- mice had greater E-C coupling failure following repeated contractions compared to fibers from mdx mice. Moreover, protease inhibition during these contractions was sufficient to attenuate E-C coupling failure in muscle fibers from both mdx and mdx/utr-/- mice. Study two evaluated the effects of overexpressing the Ca2+ buffering protein sarcoplasmic/endoplasmic reticulum Ca2+-ATPase 1 (SERCA1) in skeletal muscles from mdx and mdx/utr-/- mice. Overall, SERCA1 overexpression decreased muscle damage and protected the muscle from contraction-induced injury in mdx and mdx/utr-/- mice. In study three, the cellular mechanisms underlying the beneficial effects of SERCA1 overexpression in mdx and mdx/utr-/- mice were investigated. SERCA1 overexpression attenuated calpain activation in mdx muscle only, while partially attenuating the degradation of the calpain target desmin in mdx/utr-/- mice. Additionally, SERCA1 overexpression decreased the SERCA-inhibitory protein sarcolipin in mdx muscle but did not alter levels of Ca2+ regulatory proteins (parvalbumin and calsequestrin) in either dystrophic model. Lastly, SERCA1 overexpression blunted the increase in endoplasmic reticulum stress markers Grp78/BiP in mdx mice and C/EBP homologous protein (CHOP) in mdx and mdx/utr-/- mice. Overall, findings from the studies presented in this dissertation provide new insight into the role of Ca2+ in muscle dysfunction and damage in different dystrophic mouse models. Further, these findings support the overall strategy for improving intracellular Ca2+ control for the development of novel therapies for DMD.

Modeling the Network of Dutch and Flemish Print Production, 1550-1750

The production of artistic prints in the sixteenth- and seventeenth-century Netherlands was an inherently social process. Turning out prints at any reasonable scale depended on the fluid coordination between designers, platecutters, and publishers; roles that, by the sixteenth century, were considered distinguished enough to merit distinct credits engraved on the plates themselves: invenit, fecit/sculpsit, and excudit. While any one designer, plate cutter, and publisher could potentially exercise a great deal of influence over the production of a single print, their individual decisions (Whom to select as an engraver? What subjects to create for a print design? What market to sell to?) would have been variously constrained or encouraged by their position in this larger network (Who do they already know? And who, in turn, do their contacts know?) This dissertation addresses the impact of these constraints and affordances through the novel application of computational social network analysis to major databases of surviving prints from this period. This approach is used to evaluate several questions about trends in early modern print production practices that have not been satisfactorily addressed by traditional literature based on case studies alone: Did the social capital demanded by print production result in centralized, or distributed production of prints? When, and to what extent, did printmakers and publishers in the Low countries favor international versus domestic collaborators? And were printmakers under the same pressure as painters to specialize in particular artistic genres? This dissertation ultimately suggests how simple professional incentives endemic to the practice of printmaking may, at large scales, have resulted in quite complex patterns of collaboration and production. The framework of network analysis surfaces the role of certain printmakers who tend to be neglected in aesthetically-focused histories of art. This approach also highlights important issues concerning art historians’ balancing of individual influence versus the impact of longue durée trends. Finally, this dissertation also raises questions about the current limitations and future possibilities of combining computational methods with cultural heritage datasets in the pursuit of historical research.

The Impact of Motor Learning on Motor Behavior and Cortical Dynamics in a Complex Stressful Social Environment

An economy of effort is a core characteristic of highly skilled motor performance often described as being effortless or automatic. Electroencephalographic (EEG) evaluation of cortical activity in elite performers has consistently revealed a reduction in extraneous associative cortical activity and an enhancement of task-relevant cortical processes. However, this has only been demonstrated under what are essentially practice-like conditions. Recently it has been shown that cerebral cortical activity becomes less efficient when performance occurs in a stressful, complex social environment. This dissertation examines the impact of motor skill training or practice on the EEG cortical dynamics that underlie performance in a stressful, complex social environment. Sixteen ROTC cadets participated in head-to-head pistol shooting competitions before and after completing nine sessions of skill training over three weeks. Spectral power increased in the theta frequency band and decreased in the low alpha frequency band after skill training. EEG Coherence increased in the left frontal region and decreased in the left temporal region after the practice intervention. These suggest a refinement of cerebral cortical dynamics with a reduction of task extraneous processing in the left frontal region and an enhancement of task related processing in the left temporal region consistent with the skill level reached by participants. Partitioning performance into ‘best’ and ‘worst’ based on shot score revealed that deliberate practice appears to optimize cerebral cortical activity of ‘best’ performances which are accompanied by a reduction in task-specific processes reflected by increased high-alpha power, while ‘worst’ performances are characterized by an inappropriate reduction in task-specific processing resulting in a loss of focus reflected by higher high-alpha power after training when compared to ‘best’ performances. Together, these studies demonstrate the power of experience afforded by practice, as a controllable factor, to promote resilience of cerebral cortical efficiency in complex environments.

The Ecology of Urbanization: A Study of Soil Microbial Community Rosponse

Urbanization is associated with global biodiversity loss of macrophauna and flora through direct and indirect mechanisms, but to date few studies have examined urban soil microbes. Although there are numerous studies on the influence of agricultural management on soil microbial community composition, there has been no global-scale study of human control over urban soil microbial communities. This thesis extends the literature of urban ecology to include soil microbial communities by analyzing soils that are part of the Global Urban Soil Ecology and Education Network (GLUSEEN). Chapter 1 sets the context for urban ecology; Chapters 2 addresses patterns of community assembly, biodiversity loss, and the phylogenetic relationships among community members; Chapter 3 addresses the metabolic pathways that characterize microbial communities existing under different land-uses across varying geographic scales; and Chapter 4 relates Chapter 2 and 3 to one another and to evolutionary theory, tackling assumptions that are particular to microbial ecology.