MAG and myelin block the ability of BDNF to overcome inhibition of axonal regeneration by inhibiting BDNF's activation of Rap1
Year of Dissertation:
2009
It is well established that axons of the adult mammalian CNS do not regenerate if injured by trauma, or if afflicted by various neurodegenerative conditions. It is also well established that myelin, the insulating and protective membranous sheath around neurons, contains several proteins that act as inhibitors of neurite outgrowth, such as MAG, and damaged myelin is one of the factors limiting CNS regeneration after injury.
Functional Genomics of Cell Wall Biogenesis in Fungi
Author:
Edimarlyn Gonzalez
Year of Dissertation:
2009
In pathogenic fungi, glycosylphosphatidyl inositol cell wall proteins (GPI-CWPs) mediate adhesion of the fungus to a host during infection -a step that is necessary for the fungus to be able to initiate and establish infection. Genes involved in the biosynthetic pathway that incorporate GPI-CWPs to the wall thus represent excellent targets for antifungal drug design. A mechanistically-based genomic screen was developed and applied to search for genes involved in fungal cell wall biogenesis. Specifically, the screen identifies genes required for the cross-linking of glycoproteins (GPI-CWPs) to cell wall polysaccharides via GPI anchors. GPI-CWPs are the most abundant class of proteins at the wall and play both structural and biosynthetic functions.
MyTH4 and FERM Have Overlapping and Distinct Roles in the Function of Myo1, a Class XIV Myosin in Tetrahymena thermophila
Year of Dissertation:
2011
MyTH4 and FERM are conserved tail domains in myosin classes VII, X, XII, XIV, XV and in MyoG. Myo1, a class XIV myosin in Tetrahymena thermophila, contains MyTH4 and FERM. Previous studies have shown that Myo1 localizes to phagosomes, the cytoskeleton, and the macronucleus, and that phagosome trafficking and division of the macronucleus are affected in a MYO1 knockout. To investigate the roles for MyTH4 and FERM in the function of Myo1, GFP-tagged MyTH4, FERM, and truncated FERM were separately overexpressed in Tetrahymena. Actin antibody coprecipitated tubulin, GFP-MyTH4, and GFP-FERM. GFP-MyTH4 and GFP-FERM cosedimented with either exogenous microtubules or exogenous F-actin. GFP-MyTH4 localized to phagosomes and colocalized with antitubulin to intranuclear microtubules. Overexpression of GFP-MyTH4 inhibited the organization of the parallel array of intranuclear microtubules that form prior to division of the macronucleus. Cells that failed to form the parallel array of microtubules did not advance in nuclear division. Overexpression of GFP-MyTH4 did not affect phagosome recycling. Overexpressed GFP-FERM localized to phagosomes, cytoskeleton, and intranuclear puncta and did not affect division of the macronucleus. Overexpression of truncated GFP-FERM did not localize to the cytoskeleton or nucleus and led to the accumulation of phagosomes at the membrane recycling site in the posterior of the cell. It is unlikely that the overexpression phenotypes are nonspecific effects of GFP. Localization of GFP-fusions is consistent with the localization of full-length Myo1, and overexpression phenotypes mimic the knockout phenotype. Furthermore, GFP-MyTH4 from Myo9, another Tetrahymena myosin, did not localize in Tetrahymena thermophila. We conclude that MyTH4 and FERM have overlapping roles as indicated by the interaction with actin and tubulin. However, MyTH4 and FERM appear to have distinct roles in the function of Myo1. MyTH4 affects the organization of microtubules involved in macronuclear division, whereas FERM affects recycling of phagosomes and is required for localization to the cytoskeleton.
IMPACTS OF HABITAT DEGRADATION ON FUNDULUS HETEROCLITUS (LINNAEUS) IN URBAN TIDAL SALT MARSHES IN NEW YORK
Year of Dissertation:
2009
Despite considerable improvements in water quality over the last few decades, the ecological integrity of benthic habitats in the Arthur Kill (AK), New York, USA, largely remains altered. This dissertation explores how altered ecological status of benthic habitats directly and indirectly (via food webs) affected a resident forage fish, mummichogs (Fundulus heteroclitus), in highly urbanized tidal salt marshes in AK. A substantial portion of total abundance and biomass of benthic macroinvertebrates (a primary prey resource for mummichogs) in AK were comprised of only a few opportunistic oligochaete and polychaete species. And these alterations in benthic macroinfaunal communities in AK were strongly associated with the sediment-associated mercury level. Alterations in the AK benthic macroinfaunal assemblages were generally reflected in diet habits and strategies of mummichogs; a generalized feeding strategy with a broad diet niche breadth of mummichogs shifted to more specialized strategies for many of the AK populations. Although decapods (especially Palaemonetes spp.) were the predominant prey for all populations of mummichogs, the length-specific maximum sizes of Palaemonetes spp. ingested by some of the AK populations of mummichogs were about 2-fold smaller than those ingested by the reference population. These shifts in feeding habits were compensated for with an increased consumption of polychaetes by most of the AK populations and polychaetes contributed up to more than 40% of their gut contents. Partial trophic decoupling between mummichogs and dominant benthic macroinvertebrates had further implications for biogeochemical cycling of trace metals and energy transfer in AK. Alterations in benthic macroinfaunal prey communities reduced trophic transfer efficiency (i.e., exposure levels) of metals to mummichogs. Furthermore, despite their compensatory food consumption, most of the AK populations of mummichogs had considerably elevated total metabolism, resulting in substantially reduced growth conversion efficiency. This reduction in energy conversion efficiency at the individual level can cascade through trophic chains, potentially leading to energetic bottlenecks at the community level. Altered salt marsh trophic structures in AK and their resultant impacts on mummichog (a crucial trophic link in urban estuaries) bioenergetics may thus disrupt energy translocation in this severely degraded coastal ecosystem.
Gender specific changes in key regulators of neurodevelopment and autistic behavioral pathology in mice exposed to water chlorination byproducts
Author:
Sara Rose Guariglia
Year of Dissertation:
2010
Autism is a heterogeneous group of disorders with no definitive etiology. Out of concern for higher than expected prevalence, the Agency for Toxic Substances and Disease Registry (ATSDR) investigated the municipal water supply in Brick Township, New Jersey. The ATSDR found that two trihalomethanes (THMs), specifically chloroform and bromoform, as well as tetrachloroethylene (perchlorethylene; PCE), were present in concentrations that exceeded allowable maximum contaminant (MCL) values. In a related study, it was found that THMs and PCE act synergistically to increase the level of catalytic Protein Kinase A (PKA) in neurons of clam embryos. PKA is a key regulator of neurodevelopment, and it is hypothesized that abnormalities in PKA activity could induce both histopathological and biochemical manifestations that are found in autism. Based upon these findings, we hypothesized that THM/PCE exposure induces changes in key regulators of neurodevelopment and behavioral pathology similar to that which is found in autism. In our experiments we found that exposure to THM/PCE induces an increase in the level of catalytically active PKA in zebrafish neurons and increases PKA activity in microglia cell culture. In a mouse model, we found that exposure to THM/PCE via drinking water induces an increase in the activity of PKA in the cerebral cortex of male animals at postnatal day 4 (P4) and postnatal day 10 (P10). Females cortical PKA activity was unaffected by THM/PCE exposure. By P15, male cortical PKA activity is no longer affected by THM/PCE exposure and female cortical PKA activity remains unaffected. Behaviorally, we found that the THM/PCE exposed males develop autistic like behavioral pathology as they evidence deficits in communication and social behavior and demonstrate both perseverance behavior and anxiety. Again, this finding is gender specific, as female behavior is unaffected by THM/PCE exposure. These findings suggest that these chemicals may be involved in the etiology of autism and that males are more susceptible to this set of insults.
SYSTEMATICS AND HISTORICAL BIOGEOGRAPHY OF AGKISTRODON CONTORTRIX AND AGKISTRODON PISCIVORUS
Year of Dissertation:
2011
Many studies have revealed that lineages currently inhabiting formerly glaciated areas were pushed into southern glacial refugia and have expanded into their modern range since the last glacial maximum. There have been few studies that compare the effects of glacial cycles on lineage diversification, historical demography and migration rates in closely related species with overlapping ranges. In this study I compare phylogeographic structure, historical demography, approximate lineage age, potential distributions, and migration rates in two closely related and broadly co-occurring venomous snakes in eastern North America, the cottonmouth (Agkistrodon piscivorus) and copperhead (A. contortrix) using multilocus coalescent approaches. It has recently been discovered that gene flow between closely related species with adjacent distributions may be common (Nosil 2008). However, the absence of gene flow is a primary assumption of many phylogeographic methods including species tree inference and Bayesian species delimitation. I provide a framework for examining species delimitation when gene flow between species is present and provide a taxonomic revision of A. contortrix and A. piscivorus. In addition, I explore whether hybrids between adjacent species inhabit unique environmental conditions not suitable to one or both species. Finally, I reveal that species diversification was likely a direct result of Pleistocene glacial cycles and that species with the closest proximity to formerly glaciated areas experienced population expansion following the retreat of the Laurentide Ice Sheet. A combination of population expansion out of refugia and niche expansion has resulted in hybridization between adjacent species where species distributions come into contact. It is not clear whether gene flow has persisted during speciation and subsequent interglacial periods or if it has only recently occurred following the last glacial maximum.
PHYLOGENETICS AND BIOGEOGRAPHY OF MOUSE OPOSSUMS (DIDELPHIDAE: MARMOSA)
Year of Dissertation:
2012
This research focused on the systematics and biogeography of mouse opossums of the genus Marmosa (Mammalia: Didelphimorphia: Didelphidae), with special emphasis on species found in the complicated geography of north-central South America. Via the four chapters of this dissertation, I presented information obtained through fieldwork, examination of museum specimens, DNA sequencing, phylogenetic analyses, georeferencing (including consultation of field notes and collectors), and ecological niche modeling. In Chapter 1, I conducted phylogenetic analyses of species of Marmosa based on sequence data of the mitochondrial cytochrome-b gene (CYTB), in part to test the monophyly of species previously recognized based on morphological criteria. This study revealed the existence of unrecognized species and identified novel interspecific relationships. All trans-Andean species of the subgenus Marmosa were recovered as a clade, suggesting that the uplift of the Andes might have played an important role in the diversification of the genus. In Chapter 2, I documented the presence of M. waterhousei in the Venezuelan Andes. This finding implied that the species might have crossed the dry Depresión del Táchira during a glacial period. In Chapter 3, I investigated the phylogeography of M. robinsoni, a species predominately distributed across the dry forests of northern South America. I conducted phylogenetic analyses based on sequence data of one mitochondrial and one nuclear gene. The results confirmed the monophyly of a dry-forest clade formed by M. robinsoni and M. xerophila and showed the existence of two major clades within M. robinsoni that corresponded roughly to an east/west division. Results of ancestral area reconstructions identified multiple dispersal events out of the greater Maracaibo basin. Lastly, in Chapter 4 I used ecological niche modeling to test the geographic predictions of competition between a sister species pair, M. robinsoni and M. xerophila. The results strongly suggest that M. xerophila may isolate populations of M. robinsoni in the Península de Paraguaná of northern Venezuela--representing a novel example of geographic isolation caused by competition. Together, these studies contributed to a better understanding of the taxonomy, phylogenetics, and biogeography of the genus Marmosa; provide novel information relevant to the biogeography of dry-forest species in northwestern South America; and propose a refinement of the concept of ecological vicariance to incorporate the possibility that biotic interactions could lead to geographic isolation.
Evolutionary Analyses on the Core Genome of Borrelia burgdorferi: Elucidating the Genomics of Virulence
Year of Dissertation:
2011
ABSTRACT: Adaptive Evolution in Borrelia burgdorferi
cAMP and Polyamines Overcome Inhibition by MAG by Activating Cdk5 via Increased Expression of p35 Regulated by Activation of eIF5A
Year of Dissertation:
2010
Damaged axons in adult mammalian central nervous system (CNS) are unable to regenerate after injury although axons in the peripheral nervous system (PNS) or embryonic CNS can. The inhibitory molecules associated with myelin are one of the major obstacles to successful axon regeneration in the adult mammalian CNS. To date, three inhibitors of regeneration have been identified in myelin: NogoA, myelin-associated glycoprotein (MAG), and oligodendrocyte-myelin glycoprotein (OMpg) (Filbin, 2003). Interestingly, all these three ligands bind to the same receptor Nogo receptor (NgR) to mediate the inhibitory effect. p75NTR or TROY and Lingo-1( LRR and Ig domain-containing, Nogo Receptor interacting protein) are necessary components of the receptor complex as NgR is glycosyl phosphatidylinositol (GPI)-anchored and lacks a signaling domain (Wang et al., 2002a; Mi et al., 2004; Park et al., 2005; Shao et al., 2005). Activation of the receptor complex by myelin inhibitors activates the small GTPase RhoA resulting in rearrangement of the cytoskeleton and inhibition of axonal outgrowth (Hu and Strittmatter, 2004).
Adaptive Plasticity in the Human Saccade System
Year of Dissertation:
2012
The rapid point-to-point movements of the eyes called saccades are some of the most commonly made by humans, yet differ from nearly every other type of motor output in that they are completed too quickly to be adjusted during their execution by visual feedback. Yet, saccadic accuracy remains quite high over a lifetime despite inevitable changes to the physical structures controlling the eyes, indicating that the oculomotor system actively monitors and adjusts motor commands to achieve this consistent behavioral production. Indeed, it seems that beyond the ability to compensate for slow, age-related bodily changes, saccades can be modified following traumatic injury or pathology that affects their production, or in response to more short-term systematic alterations to post-saccadic visual feedback in a lab setting. It is, in fact, thought that all of these forms of plasticity rely on the visual detection of accuracy errors by a unified set of mechanisms that support the process known as saccade adaptation. A great deal has been learned about saccade adaptation, as it has been extensively studied as a phenomenon in its own right, as well as being used to explore the process of motor learning in general. However, many fundamental questions about saccade adaptation remain unanswered, often related to the way that saccade adaptation might operate in the natural environment with substantially more complex visual stimuli than are generally used in the lab. Here, we addressed these questions with (in some cases original) variants and more conventional examples of the frequently used intrasaccadic target step (ISS) paradigm (in which an experimenter causes saccadic error by shifting a target during the movement). By exploring the responses to whole-field-ISSs, we have inferred that saccade adaptation might be supported by a trans-saccadic integration mechanism, and may be sensitive to intrasaccadic motion signals. Challenging the oculomotor system by confronting it with multiple post-saccadic targets has revealed that saccade adaptation can occur in a target-identity specific manner, so that even if post-saccadic error varies from trial-to-trial, adaptation seems to reflect the average behavior of the target. At a more basic level, we systematically varied ISSs to determine the lower limits of the oculomotor system's sensitivity to intrasaccadic displacement during adaptation. Also at a more basic level, we looked at the effects of rendering post-saccadic feedback more intermittent during adaptation, finding it to have little effect on the magnitude or rate of adaptive dynamics, similar to other forms of motor learning but somewhat dissimilar from operant conditioning. These experiments also furnished a useful setting to develop and test a novel model of saccade adaptation which more explicitly relies on post-saccadic sensory prediction than previous models, but that is nonetheless in keeping with the ethos of modern motor learning theory. Finally, we found that the establishment and maintenance of a context for saccadic performance by adaptation could be achieved by consistently pairing a target's visual-identity with a specific ISS, extending what had been previously recognized as constituting a cue for contextual motor learning. In general, our results suggest that saccade adaptation is a highly flexible mechanism that not only supports the maintenance of accuracy, but also makes use of a wide range of brain functions to deftly tailor saccadic behavior contingent on task demands.