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Isoform-Specific Roles Of Extracellular Signal-Regulated Kinases In Pain, Benedict Alter
Isoform-Specific Roles Of Extracellular Signal-Regulated Kinases In Pain, Benedict Alter
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ABSTRACT OF THE DISSERTATION Isoform-specific roles of Extracellular Signal-Regulated Kinases in pain by Benedict Joseph Alter Doctor of Philosophy in Biology and Biomedical Sciences Neurosciences Washington University in St. Louis, 2012 Professor Robert Gereau, Chairperson The extracellular signal-regulated kinase: ERK) isoforms, ERK1 and ERK2, are believed to be key signaling molecules in nociception and nociceptive sensitization. Studies utilizing inhibitors targeting the shared ERK1/2 upstream activator, mitogen-activated protein kinase kinase: MEK), and transgenic mice expressing a dominant negative form of MEK have established the importance of ERK1/2 signaling. However, these techniques do not discriminate between ERK1 and ERK2. To dissect the …
From Bacteria To Human: Biophysical Studies Of Inward Rectifying Potassium Channels, Wayland Cheng
From Bacteria To Human: Biophysical Studies Of Inward Rectifying Potassium Channels, Wayland Cheng
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Inward rectifying potassium: Kir) channels are important in regulating cellular excitability in organs such as the heart, brain and pancreas. Prokaryotic KirBac channels are structurally similar to eukaryotic Kir channels, but distantly related and of unknown function. The goal of this thesis has been to investigate the functional properties of KirBac1.1 and to relate these findings to eukaryotic Kir channels. The approach was to use recombinantly-expressed, purified K+ channels--KirBac1.1, KcsA, Kir2.1 and Kir2.2--in order to integrate findings from functional studies, using liposomal flux assays and patch-clamping, with high-resolution crystal structures. By reconstituting KirBac1.1 into giant liposomes for patch-clamping, I show …
Joint Functions Of Mett-10 And Dynein In The Caenorhabditis Elegans Germ Line, Maia Dorsett
Joint Functions Of Mett-10 And Dynein In The Caenorhabditis Elegans Germ Line, Maia Dorsett
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During normal development as well as in diseased states such as cancer, extracellular "niches" often provide cues to proximal cells and activate intracellular pathways. Activation of such signaling pathways in turn instructs cellular proliferation and differentiation. In the C.elegans gonad, GLP-1/Notch signaling instructs germ line stem cells to self renew through mitotic cell division. As germ cells progressively move out of the niche, they differentiate by entering meiosis and eventually form gametes. Using this model system, I uncovered a cooperative role for the METT-10 putative methyltransferase and the dynein motor complex in regulating the balance between germ cell proliferation and …
Atm Deficiency -- A Multifaceted Defect In Lymphocyte Development, Beth Helmink
Atm Deficiency -- A Multifaceted Defect In Lymphocyte Development, Beth Helmink
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The generation of a functional antigen receptor gene in developing lymphocytes requires that the second exon be assembled through a process known as V(D)J recombination, a process that necessarily involves the generation and repair of DNA double-strand breaks made by the Rag endonuclease. Double strand breaks incurred during G1 of the cell cycle activate ATM, a PI3-kinase-like kinase that, in response to genotoxic DNA damage, is known to phosphorylate hundreds of proteins with unique and diverse functions. Notably, deficiencies in ATM lead to ataxia-telangiectasia, a syndrome characterized by lymphopenia, genomic instability and a predisposition to tumors involving antigen receptor loci …
The Role Of Autophagy And Il-17 In Bone Resorption, Carl Deselm
The Role Of Autophagy And Il-17 In Bone Resorption, Carl Deselm
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Osteoclasts are essential for skeletal homeostasis: Teitelbaum, 2000). These macrophage-lineage cells function by generating a polarized microenvironment between themselves and bone wherein skeletal matrix is degraded. This resorptive compartment is isolated from the general extracellular space by an actin ring which encompasses the ruffled border, a convoluted plasma membrane structure formed by its fusion with lysosome-related vesicles containing an electrogenic H+ATPase, a chloride channel, LAMP-1, and cathepsin K. In consequence, resorption of bone reflects secretion of HCl to mobilize mineral, and cathepsin K to degrade the collagen-rich organic matrix, into the resorptive space: Stenbeck, 2002; Zhao et al., 2008). We …
The Roles Of Vasoactive Intestinal Polypeptide In Circadian Entrainment Of Suprachiasmatic Nucleus, Sungwon An
The Roles Of Vasoactive Intestinal Polypeptide In Circadian Entrainment Of Suprachiasmatic Nucleus, Sungwon An
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In mammalian hypothalamus, the suprachiasmatic nucleus: SCN) generates daily behavioral and physiological rhythms as a circadian pacemaker. The 20,000 SCN neurons synchronize to each other and to the ambient cues to generate coherent daily rhythms. Vasoactive intestinal polypeptide: VIP), a neuropeptide produced by SCN neurons, plays a major role in synchronizing SCN neurons to each other. Whether VIP mediates synchrony to environmental cues and how synchrony within the SCN is achieved has not been examined extensively. We recorded PERIOD::LUCIFERASE: PER2::LUC) expression from SCN explant cultures over multiple days following VIP application at different circadian time points to generate a phase …
Regulation Of Neutrophil Homeostasis By Chemokines Signaling Through The Cxcr2 And Cxcr4 Receptors, Kyle Eash
Regulation Of Neutrophil Homeostasis By Chemokines Signaling Through The Cxcr2 And Cxcr4 Receptors, Kyle Eash
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The number of neutrophils in the blood is tightly regulated to ensure adequate protection against microbial pathogens while minimizing damage to host tissue. Neutrophil homeostasis in the blood is achieved through a balance of neutrophil production, release from the bone marrow, and clearance from the circulation. Accumulating evidence suggests that chemokine signaling in the bone marrow may play a key role in maintaining neutrophil homeostasis. Based on this evidence, we developed a "tug-of-war" model in which opposing chemokine gradients, specifically release-inducing CXCR2 signals and retention-promoting CXCR4 signals, act antagonistically to regulate neutrophil release from the bone marrow. We generated mice …
The Manipulated Mechanism: Towards An Account Of The Experimental Discovery Of Mechanistic Explanations, Donald Goodman-Wilson
The Manipulated Mechanism: Towards An Account Of The Experimental Discovery Of Mechanistic Explanations, Donald Goodman-Wilson
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Recent work in the philosophy of biology has sought after an account of mechanistic explanation. Biologists frequently encounter causal relationships that beg for explanation. For example, genes appear to encode for particular phenotypes. How does gene expression work? Biologists posit mechanisms to explain the link between cause and effect. Thus, gene expression would be explained by an appeal to a complex mechanism linking the gene to the phenotype, as such an appeal will provide answers to broad ranges of "how" and "why" questions about the causal relationship, and predict novel effects. Here, I focus on a recent problem raised for …
Mhc Class I-Dependent Acquisition And Maintenance Of Natural Killer Cell Function, Julie Chase
Mhc Class I-Dependent Acquisition And Maintenance Of Natural Killer Cell Function, Julie Chase
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Natural killer: NK) cells are hematopoietically derived immune cells of the lymphoid lineage. Their effector functions, including cytokine secretion and cytotoxicity, are controlled by the integration of signals received from both activating and inhibitory receptors. In a phenomenon known as "missing-self" recognition, NK cells eliminate self-tissues with aberrantly low or absent MHC class I surface expression, as is common in settings of viral infection and transformation, when these cells fail to engage the critical NK cell inhibitory receptors. Interestingly, NK cells that develop in an MHC class I-deficient environment are defective in natural killing and hyporesponsive to stimulation through their …
The Evolution And Reproductive Ecology Of Oenothera (Onagraceae), Kyra Krakos
The Evolution And Reproductive Ecology Of Oenothera (Onagraceae), Kyra Krakos
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This dissertation describes the role of pollination in the floral diversification of Oenothera with an integration of both ecological and phylogenetic approaches. Oenothera: Onagraceae) is a model system for studying plant reproductive biology. It provides excellent examples of shifts in reproductive traits such as pollination and breeding system, features that have been important in angiosperm diversification. These systems are evolutionarily labile; they easily shift between different states. These different reproductive traits may shift in a concerted fashion; therefore, a more comprehensive approach to understanding the evolution of these plant systems simultaneously addresses shifts in pollination and breeding system. Using 54 …
Effects Of The Aquatic To Terrestrial Habitat Ratio On An Amphibian Predator And Its Prey, Amber Kramer
Effects Of The Aquatic To Terrestrial Habitat Ratio On An Amphibian Predator And Its Prey, Amber Kramer
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This dissertation explores the effects of varying the composition of aquatic and terrestrial habitats in a landscape: the aquatic to terrestrial ratio) on amphibians that use both the aquatic and terrestrial habitats during their lives. In Chapter 1, I first used meta-analysis and simulations to demonstrate that as the longevity of an amphibian increases, the elasticity of the population growth rate to perturbations in the aquatic: larval) habitat decreases. In Chapter 2, I examined the abundance of larvae of a long-lived amphibian, Ambystoma maculatum, across landscapes that varied in their aquatic to terrestrial ratios and found that larvae of this …
Interactions Between Bmp And Canonical Wnt Signaling Regulate Critical Stages Of The Osteoblast Lifecycle, Valerie Salazar
Interactions Between Bmp And Canonical Wnt Signaling Regulate Critical Stages Of The Osteoblast Lifecycle, Valerie Salazar
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Skeletal development and post-natal bone homeostasis are dependent on the coordinated activity of bone-forming cells called osteoblasts and bone-resorbing cells called osteoclasts. Over 10 million people in the US currently suffer from osteoporosis, which increases the risk of low-trauma fractures and accounts for health care expenditures exceeding $14 billion per year. Osteoporosis is often treated with anti-resorptive compounds, primarily bisphosphonates, which inhibit osteoclast-mediated bone destruction. However, these drugs do not restore bone mass, which can only be accomplished by activation new bone formation, as with intermittent parathyroid hormone therapy. The development of additional bone "anabolic" therapies will require genetic and …
The Role Of The Membrane Proximal Region Of The M2 Cytoplasmic Tail In Virus Replication, Shaun Stewart
The Role Of The Membrane Proximal Region Of The M2 Cytoplasmic Tail In Virus Replication, Shaun Stewart
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Influenza A virus encodes M2, a proton channel that has been shown to be important during virus entry and assembly. The primary aim of this thesis was to investigate the role of the membrane proximal region, residues 46-69, of the M2 cytoplasmic tail during virus replication. A cholesterol recognition/interaction amino consensus: CRAC) motif, previously identified in the membrane proximal region of M2 in some influenza A virus strains, was suggested to play a role in virus replication by mediating incorporation of M2 into budding virus particles. Alteration or completion of the M2 CRAC motif in two different recombinant virus strains …
Molecular Genetic Analysis Of Non-Catalytic Pol Iv And V Subunits, Ek Han Tan
Molecular Genetic Analysis Of Non-Catalytic Pol Iv And V Subunits, Ek Han Tan
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Among eukaryotes, plants have the distinction of encoding multisubunit RNA polymerases used exclusively for RNA directed DNA Methylation: RdDM) in addition to Pol I, II, and III. In Arabidopsis thaliana, Pol IV is required for the biogenesis of 24nt siRNAs whereas Pol V transcription is needed for cytosine methylation of the DNA sequences corresponding to these siRNAs. The ancestry of Pol IV and V can be traced back to Pol II, and Pol II, IV and V still utilize multiple non-catalytic subunits encoded by the same genes. Genetic analysis of non-catalytic subunits that are highly similar reveals that these subunits …
The Association Between Traumatic Brain Injury And Alzheimer's Disease: Mouse Models And Potential Mechanisms, Hien Tran
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Alzheimer's disease: AD) is a neurodegenerative disorder characterized pathologically by progressive neuronal loss, extracellular plaques containing the amyloid-β: Aβ) peptides, and neurofibrillary tangles: NFTs) composed of hyperphosphorylated tau proteins. Aβ is thought to act upstream of tau, affecting its phosphorylation and therefore aggregation state. One of the major risk factors for AD is traumatic brain injury: TBI). Acute intra-axonal Aβ and diffuse extracellular plaques occur in approximately 30% of human subjects following severe TBI. Intra-axonal accumulations of total and phospho-tau and less frequently NFTs have also been found in these patients. Due to the lack of an appropriate small animal …
The Gut Microbiome In Healthy And Severely Malnourished Humans, Tanya Yatsunenko
The Gut Microbiome In Healthy And Severely Malnourished Humans, Tanya Yatsunenko
All Theses and Dissertations (ETDs)
Human large intestine is home to tens of trillions of microbes belonging to all three do- mains of life. The functions encoded by the genes in this community: microbiome) include processing and production of macro- and micronutrients. Much remains unknown about the factors that determine the assembly of the gut microbial community starting at birth, and if disruptions in the assembly of this `microbial metabolic organ' early in life result in physiologic and metabolic deficits later in life. The central goal of my thesis was to char- acterize development of the gut microbiome early in life, with a focus on …
Discovering Conserved Cis-Regulatory Elements That Regulate Expression In Caenorhabditis Elegans, Nnamdi Ihuegbu
Discovering Conserved Cis-Regulatory Elements That Regulate Expression In Caenorhabditis Elegans, Nnamdi Ihuegbu
All Theses and Dissertations (ETDs)
The aim of this dissertation is two-fold:: 1) To catalog all cis-regulatory elements within the intergenic and intronic regions surrounding every gene in C.elegans: i.e. the regulome) and: 2) to determine which cis-regulatory elements are associated with expression under specific conditions. We initially use PhyloNet to predict conserved motifs with instances in about half of the protein-coding genes. This initial first step was valuable as it recovered some known elements and cis-regulatory modules. Yet the results had a lot of redundant motifs and sites, and the approach was not efficiently scalable to the entire regulome of C. elegans or other …
The Functions Of Human Dna2 In Mitochondrial And Nuclear Dna Maintenance, Julien Duxin
The Functions Of Human Dna2 In Mitochondrial And Nuclear Dna Maintenance, Julien Duxin
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Coordination between DNA replication, DNA repair and cell-cycle progression ensures high fidelity DNA replication thus preventing mutations and DNA rearrangements. Interestingly, in addition to nuclear DNA stability, mitochondrial DNA: mtDNA) integrity is also essential for normal development. The current challenge resides in unraveling the different mechanisms that govern nuclear and mtDNA stability and to understand how these two separated genomes have evolved. This work focuses on delineating the biological functions of human Dna2: hDna2). Dna2 is a highly conserved helicase/nuclease that in yeast participates in DNA replication and Okazaki fragment maturation, DNA repair, and telomere maintenance. Immunofluorescence and biochemical fractionation …
Protein-Dna Recognition Models For The Homeodomain And C2h2 Zinc Finger Transcription Factor Families, Ryan Christensen
Protein-Dna Recognition Models For The Homeodomain And C2h2 Zinc Finger Transcription Factor Families, Ryan Christensen
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Transcription factors: TFs) play a central role in the gene regulatory network of each cell. They can stimulate or inhibit transcription of their target genes by binding to short, degenerate DNA sequence motifs. The goal of this research is to build improved models of TF binding site recognition. This can facilitate the determination of regulatory networks and also allow for the prediction of binding site motifs based only on the TF protein sequence. Recent technological advances have rapidly expanded the amount of quantitative TF binding data available. PBMs: Protein Binding Microarrays) have recently been implemented in a format that allows …
Mechanism Of Yeast Prion Portein Aggregation And Strain Formation, Tejas Baba Kalastavadi
Mechanism Of Yeast Prion Portein Aggregation And Strain Formation, Tejas Baba Kalastavadi
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Misfolding and aggregation of the prion protein: PrP) causes fatal neurodegenerative diseases in many mammalian species, including humans. Mutations in the gene encoding PrP are associated with ~15% of the incidences, while, the vast majority of the cases are sporadic. Interestingly, prion diseases also display pathological variation, suggesting that there are multiple different strains. To elucidate the mechanism of prion protein aggregation and strain formation, I have taken advantage of the yeast prions [PSI+] and [RNQ+] and their protein determinants Sup35p and Rnq1p, respectively. Using a Sup35-PrP chimera, I have investigated the effect of the disease associated oligopeptide repeat domain: …
Genetic Analysis Of The Pi3k/Akt/Mtor Signaling Pathway, Janna Hutz
Genetic Analysis Of The Pi3k/Akt/Mtor Signaling Pathway, Janna Hutz
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Cancer is a leading cause of human death, and it is fundamentally attributable to dysfunctional cell signaling. The PI3K/AKT/mTOR pathway is an important pro-growth intracellular signaling cascade that is often inappropriately activated in a wide array of cancers. Efforts to develop anticancer drugs have therefore focused, in part, on identifying PI3K/AKT/mTOR pathway inhibitors. However, patient response to some such inhibitors is mixed, with some patients experiencing a paradoxical activation of the pathway following treatment. It is therefore necessary to better understand the nature of the PI3K/AKT/mTOR pathway and how it varies in different individuals. The work presented here used cell …
A Search For Genetic Modifiers Responsible For Congenital Heart Disease Variability In The Presence Of Nkx2-5 Haploinsufficiency, Julia Brandeis Winston Honold
A Search For Genetic Modifiers Responsible For Congenital Heart Disease Variability In The Presence Of Nkx2-5 Haploinsufficiency, Julia Brandeis Winston Honold
All Theses and Dissertations (ETDs)
While a clear heritable risk has been observed for congenital heart disease, there is considerable variation in penetrance and presentation likely due to multiple genetic and environmental risk factors. To identify causative factors and interactions responsible for variability in heart development, greater than 4,200 hearts from Nkx2-5 heterozygous knockout mice have been collected and examined. Nkx2-5+/- mice in the inbred strain background C57Bl/6 frequently have atrial and ventricular septal defects. The incidences are substantially reduced in the Nkx2-5+/- progeny of first-generation: F1) outcrosses to the strains FVB/N or A/J. Defects recur in the second generation: F2) of the F1xF1 intercross …
Regulation Of Peripheral Nerve Regeneration By The Mtor Pathway, Namiko Abe
Regulation Of Peripheral Nerve Regeneration By The Mtor Pathway, Namiko Abe
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While neurons in the central nervous system: CNS) have limited capacity for regrowth after damage, neurons in the peripheral nervous system: PNS) have a robust ability to regenerate their axons following injury. Successful regeneration depends upon both extrinsic cues in the environment and the activation of intrinsic mechanisms to promote regrowth. A number of inhibitory molecules in the CNS environment that prevent axonal regrowth have been identified, but less is known regarding the signaling mechanisms that regulate regenerative ability in PNS neurons. Here, we explored multiple components of injury signaling in the PNS, including the retrograde transport of local axonal …
Impact Of Second Messenger Modulation On Activity-Dependent And Basal Properties Of Excitatory Synapses, Chun Yun Chang
Impact Of Second Messenger Modulation On Activity-Dependent And Basal Properties Of Excitatory Synapses, Chun Yun Chang
All Theses and Dissertations (ETDs)
Cognitive processing in the central nervous system relies on accurate information propagation; neurotransmission is the fundamental mechanism underlying network information flow. Because network information is coded by the timing and the strength of neuronal activity, synaptic properties that translate neuronal activity into synaptic output profoundly determine the precision of information transfer. Synaptic properties are in turn shaped by changes in network activity to ensure appropriate synaptic output. Activity-dependent adjustment of synaptic properties is often initiated by second messenger signals. Understanding how second messengers sculpt synaptic properties and produce changes in synaptic output is key for elucidating the interplay between network …
Regulation Of Human T-Cell Leukemia Virus Type 1 Infection And Replication, Gunjan Choudhary
Regulation Of Human T-Cell Leukemia Virus Type 1 Infection And Replication, Gunjan Choudhary
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Retroviruses have evolved complex mechanisms to regulate their cellular tropism and gene expression. It is generally accepted that productive infections proceed via interactions between viral envelope molecules and specific receptors on the host cell surface. Currently, there is no known receptor for HTLV-1, though a number of factors that enhance entry have been identified. In an effort to identify a cellular receptor or attachment factor for HTLV-1, we carried out a retroviral cDNA library screen, in which cDNA from permissive HeLa S3 cells was introduced into poorly susceptible NIH 3T3 cells. These cells were selected after infection with HTLV-1 envelope …
P38 Phoshporylates Rb On Ser567 By A Novel, Cell Cycle-Independent Mechanism That Triggers Rb-Hdm2 Interaction And Apoptosis, Rachel Delston
P38 Phoshporylates Rb On Ser567 By A Novel, Cell Cycle-Independent Mechanism That Triggers Rb-Hdm2 Interaction And Apoptosis, Rachel Delston
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The retinoblastoma protein: Rb) inhibits both cell division and apoptosis, but the mechanism by which Rb alternatively regulates these divergent outcomes remains poorly understood. Cyclin dependent kinases: Cdks) promote cell division by phosphorylating and reversibly inactivating Rb by a hierarchical series of phosphorylation events and sequential conformational changes. The stress-regulated mitogen activated protein kinase: MAPK) p38 also phosphorylates Rb, but it does so in a cell cycle-independent manner that is associated with apoptosis rather than with cell division. Here, we show that p38 phosphorylates Rb by a novel mechanism that is distinct from that of Cdks. p38 bypasses the cell …
Brain Insulin Action Regulates Hypothalamic Glucose Sensing And The Counterregulatory Response To Hypoglycemia, Kelly Diggs-Andrews
Brain Insulin Action Regulates Hypothalamic Glucose Sensing And The Counterregulatory Response To Hypoglycemia, Kelly Diggs-Andrews
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The brain is the primary organ that senses blood glucose levels and initiates a stress response when blood glucose levels are too low: hypoglycemia). Insulin-dependent people with Type 1 diabetes: T1DM) have an impaired ability to sense hypoglycemia and an impaired ability to activate this counterregulatory response: CRR) to hypoglycemia. As a result, T1DM are at a greater risk of experiencing insulin induced severe hypoglycemic episodes, which can result in seizures, brain damage, or even death. Since hypoglycemia is a major barrier that limits intensive blood glucose control, important research initiatives are needed to prevent or reduce the burden of …
Use-Dependent Plasticity Regulates Sleep Need In Drosophila Melanogaster, Jeffrey Donlea
Use-Dependent Plasticity Regulates Sleep Need In Drosophila Melanogaster, Jeffrey Donlea
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Although the necessary functions of sleep have not been identified, sleep has been shown to play an important role in the consolidation of memories. Recent studies have shown that, in addition to playing a strong role in sleep regulation, the circadian clock also influences processes associated with learning and memory. Thus, the neural circuits that control circadian rhythms are uniquely positioned to play an important role in coordinating interactions between sleep and memory. Drosophila melanogaster exhibit increased sleep following several days of social experience and require sleep to consolidate long-term memories: LTM) after Courtship Conditioning, an associative memory assay. We …
Local And Systemic Consequences Of Reducing Notch Signaling In Skin Keratinocytes, Shadmehr Demehri
Local And Systemic Consequences Of Reducing Notch Signaling In Skin Keratinocytes, Shadmehr Demehri
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Notch is a transmembrane receptor that mediates short-range signaling between neighboring cells. Notch signaling has been implicated in various cellular and developmental processes essential in the life of metazoans. Specifically, Notch signaling plays a critical role in mammalian skin. Removal of Notch alleles in skin keratinocytes has been associated with an array of phenotypes with varying severity based on the identity and number of remaining Notch receptors. Phenotypes include carcinogenesis: in the case of Notch1 loss), transformation of hair follicles to epidermal cysts and neonatal lethality, the latter seen in the absence of all Notch signaling. Although these phenotypes were …
Neurophysiological Adaptations To Resistance Training And Repetitive Grasping, Michael Falvo
Neurophysiological Adaptations To Resistance Training And Repetitive Grasping, Michael Falvo
All Theses and Dissertations (ETDs)
Perhaps the most prominent feature of the central nervous system is its ability to respond to experience and its environment. Understanding the processes and mechanisms that govern adaptive behavior provides insights into its plastic nature. Capitalizing on this plasticity is of critical importance in response to injury and recovery: 35, 106), and the importance of its promotion is increasingly recognized by rehabilitation scientists. Neurophysiological techniques permitting study of cortical function in vivo may play a significant role in validating exercise interventions and disease management approaches: 14). It may be possible that with these advances we may better understand the relationship …