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Using A Novel Multiplexing Method To Track Cell Populations And Cytodifferentiation During Development Of The Submandibular Salivary Gland, Charles Thomas Manhardt
Using A Novel Multiplexing Method To Track Cell Populations And Cytodifferentiation During Development Of The Submandibular Salivary Gland, Charles Thomas Manhardt
Legacy Theses & Dissertations (2009 - 2024)
The development of submandibular salivary glands is complex and requires coordination of specific signaling events. Submandibular salivary glands originate as an epithelial invagination into the adjacent mesenchyme that leads to a single stalk and end bud; this end bud will go through a clefting process. Numerous rounds of clefting will lead to a fully developed salivary gland by this process, which is known as branching morphogenesis. As the gland undergoes morphogenesis, specific cues leading to differentiation of multiple cell types and even epithelial sub classes are required. By the later stages of development the glands are fully innervated, have an …
Vitamin D Signaling And The Impact On Mouse Mammary Gland Epithelium, Donald Grant Matthews
Vitamin D Signaling And The Impact On Mouse Mammary Gland Epithelium, Donald Grant Matthews
Legacy Theses & Dissertations (2009 - 2024)
A strong correlation between vitamin D deficiency and increased breast cancer incidence and mortality has been well established, leading researchers to further examine the possibility of using vitamin D in treatment and prevention strategies. Our long term goal is to elucidate the mechanisms by which vitamin D pathway signaling impacts on breast cancer. Generation of the high-affinity vitamin D receptor (VDR) ligand 1,25-dihydroxyvitamin D (1,25-D) from 25-hydroxyvitamin D (25-D) is mediated by CYP27B1 which is present and developmentally regulated in multiple tissues including the mammary gland. We utilized transgenic mouse models combined with diet-induced vitamin D deficiency to determine whether …
Brown Adipogenesis Of Mouse Embryonic Stem Cells In Alginate Microstrands, Andrea Mannarino Unser
Brown Adipogenesis Of Mouse Embryonic Stem Cells In Alginate Microstrands, Andrea Mannarino Unser
Legacy Theses & Dissertations (2009 - 2024)
The ability of brown adipocytes (fat cells) to dissipate energy as heat shows great promise for the treatment of obesity and other metabolic disorders. Employing pluripotent stem cells, with an emphasis on directed differentiation, may overcome many issues currently associated with primary fat cell cultures. However, brown adipocytes are difficult to transplant in vivo due to the instability of fat, in terms of necrosis and neovascularization, once injected. Thus, 3D cell culture systems that have the potential to mimic adipogenic microenvironments are needed, not only to advance brown fat implantation, but also to better understand the role of brown adipocytes …
A Novel Method For Studying Gene Regulatory Elements In Vivo Reveals Essential Roles For Intron Splicing In Neurofilament Gene Expression, Chen Wang
Legacy Theses & Dissertations (2009 - 2024)
Axon outgrowth requires coordinated expression of critical cytoskeletal genes in response to extracellular cues. The spatial and temporal expression of these genes is regulated transcriptionally and posttranscriptionally. Such regulation is largely mediated through the activity of cis-regulatory elements within the cytoskeletal DNAs and RNAs, which recruit specific sets of trans-factors such as DNA- and RNA-binding proteins, as well as microRNAs. To identify the functionally active cis-elements, characterize their dynamically changing sets of trans-factors, and investigate the association between the two sets of regulators in the context of an intact developing nervous system, one needs to combine appropriate in vivo strategies …
Mechanisms Of Age-Related Inflammation And Cancer : The Synergistic Effect Of Oxidants And Calcium, Donald A. Mccarthy
Mechanisms Of Age-Related Inflammation And Cancer : The Synergistic Effect Of Oxidants And Calcium, Donald A. Mccarthy
Legacy Theses & Dissertations (2009 - 2024)
The accumulation of senescent cells during the process of aging has been implicated as causal in numerous age-related pathologies. Senescent cells adopt a secretory phenotype consisting of many factors including matrix remodeling enzymes, growth factors, cytokines, and chemokines. Their secretory nature is the primary reason that they are associated with disease, but it remains unclear why they become so inflammatory. Using primary human fibroblasts cultured to senescence, we mechanistically determined why senescent cells are such potent inducers of inflammation. Our findings indicate that the early production of the cytokine Interleukin 1-α (IL-1α) is central to this transition. We found that …
Mouse Salivary Gland Development Requires Optimal Substrate Compliance, Sarah Beth Peters
Mouse Salivary Gland Development Requires Optimal Substrate Compliance, Sarah Beth Peters
Legacy Theses & Dissertations (2009 - 2024)
The embryonic mouse submandibular salivary gland (mSMG) is a classic model to study mechanochemical signals involved in morphogenesis and differentiation amongst multiple, interacting cell populations. Scaffold compliance is alters cellular phenotype, but little is known about the effects of compliance on organogenesis. The overall aim of this project was to demonstrate that the processes of salivary gland epithelial branching morphogenesis and differentiation require a compliant, in vivo-like environment and are disrupted outside of this range.
Computational Modeling-Driven Study Of Signaling Pathway Affecting Cleft Formation During Submandibular Salivary Gland Development, Shayoni Ray
Legacy Theses & Dissertations (2009 - 2024)
Branching morphogenesis is a conserved mechanism of mammalian organ development (salivary glands, lungs, mammary glands, pancreas, kidney, etc.), and has been a classic topic of study for generations of developmental biologists. During the process of tissue formation, several cellular and molecular and protein signaling events occur at successive stages enabling proper formation of tissue shape and structure. In order to provide a comprehensive understanding of the process of branching morphogenesis, we need a systems level approach to integrate multifactorial information. My overall global hypothesis is that multiple mathematical modeling approaches can be utilized to extract crucial cellular and tissue level …
The Lipogenic Phenotype Of Her2/Neu-Positive Breast Cancer Cells, Jan Martin Baumann
The Lipogenic Phenotype Of Her2/Neu-Positive Breast Cancer Cells, Jan Martin Baumann
Legacy Theses & Dissertations (2009 - 2024)
Recent work has shown that HER2/neu-positive breast cancer cells rely on a unique Warburg-like metabolism for survival and aggressive behavior. These cells are dependent on fatty acid (FA) synthesis, show markedly increased levels of stored fats and disruption of the synthetic process results in apoptosis. Supplementation of the growth media with physiological concentrations of saturated FAs induces cell death, whereas HER2-normal cells are not affected. This is particularly interesting in the context of new epidemiological data showing that a diet rich in saturated FAs is positively correlated with the development of HER2-negative disease, but not HER2/neu-positive disease.
The Postsynaptic Regulation Of Synaptic Strength In Drosophila, Daniel Michael Gertner
The Postsynaptic Regulation Of Synaptic Strength In Drosophila, Daniel Michael Gertner
Legacy Theses & Dissertations (2009 - 2024)
Postsynaptic Ca2+ plays an important role in synaptic homeostasis and synaptic plasticity. Postsynaptic Ca2+ signals have been shown to regulate synaptic transmission at the Drosophila larval neuromuscular junction (NMJ), however, these signals have not been well characterized. This will explore how these signals regulate synaptic strength and what channels are involved. In previous lab experiments Ca2+ transients were observed during evoked and spontaneous release (Desai and Lnenicka, 2011). It was further demonstrated that a reduction in synaptic strength occurs following synaptic stimulation. It was hypothesized that the increase in postsynaptic Ca2+ following synaptic stimulation activates the gCS and causes a …
Molecular Actions Of The Vitamin D Receptor In Breast Cancer, Erika Laporta
Molecular Actions Of The Vitamin D Receptor In Breast Cancer, Erika Laporta
Legacy Theses & Dissertations (2009 - 2024)
1,25-Dihydroxyvitamin D (1,25D) exerts anti-cancer actions through the vitamin D receptor (VDR) but the specific targets that mediate these effects remain to be defined. In these studies, growth and genomic responses to 1,25D were evaluated in a cellular model system derived from mammary tumors generated in VDR knockout (KO) and wildtype (WT) mice. WT145 cells (derived from WT tumors) expressed VDR and were growth inhibited by 1,25D, whereas KO240 cells (derived from VDRKO tumors) lacked VDR and were not growth inhibited by 1,25D. KO240 cell clones stably expressing VDR (KOhVDR cells) were sensitized to 1,25D mediated growth arrest. Genomic profiling …
Post-Translational Modification Regulates Heterogeneous Nuclear Ribonucleoprotein K Function During Axon Outgrowth In Xenopus Laevis, Erica J. Hutchins
Post-Translational Modification Regulates Heterogeneous Nuclear Ribonucleoprotein K Function During Axon Outgrowth In Xenopus Laevis, Erica J. Hutchins
Legacy Theses & Dissertations (2009 - 2024)
The RNA-binding protein, heterogeneous nuclear ribonucleoprotein K (hnRNP K), is required for axon outgrowth. Its suppression in Xenopus embryos causes defects in the translation of mRNAs of multiple cytoskeletal genes. Studies in cell lines have established that hnRNP K shuttles between the nucleus and the cytoplasm to bind and regulate the fates of its target RNAs, from splicing to export and translation. At each step, hnRNP K is regulated through post-translational modifications that alter its nucleic acid and protein interactions, and subcellular localization. Precisely how this happens in developing neurons to coordinate cytoskeletal gene expression with the extracellular signals directing …
Characterizing The Effects Of Glutaraldehyde On The Ryanodine Receptor Calcium Release Channel, Joshua Dov Strauss
Characterizing The Effects Of Glutaraldehyde On The Ryanodine Receptor Calcium Release Channel, Joshua Dov Strauss
Legacy Theses & Dissertations (2009 - 2024)
Ryanodine receptors (RyR) are large intracellular calcium release channels, which
The Role Of An Rna Binding Protein Hnrnp K During Axon Development And Regeneration In Xenopus Laevis, Yuanyuan Liu
The Role Of An Rna Binding Protein Hnrnp K During Axon Development And Regeneration In Xenopus Laevis, Yuanyuan Liu
Legacy Theses & Dissertations (2009 - 2024)
Coordinated synthesis and assembly of the cytoskeletal network contribute significantly to morphological changes during axon outgrowth. Previous studies demonstrated that heterogeneous ribonucleoprotein K (hnRNP K), an RNA binding protein, binds to the 3'-untranslated regions of all neurofilament triplet subunits, the most abundant components of the axonal cytoskeleton. These findings raised the hypothesis that hnRNP K post-transcriptionally mediates the coordinated expression of axonal cytoskeletal components. In my thesis, I test this hypothesis during both axonal development and regeneration.
Identification Of Trophic And Transcriptional Requirements For Human Embryonic Stem Cell Culture And Guided Neural Differentiation, Nicolas Edward Tokas
Identification Of Trophic And Transcriptional Requirements For Human Embryonic Stem Cell Culture And Guided Neural Differentiation, Nicolas Edward Tokas
Legacy Theses & Dissertations (2009 - 2024)
Translational biomedical research holds potential for the development of novel therapeutics for the treatment of injury, illness or disability. To better understand intrinsic cellular and molecular biology, human embryonic stem cells (hESCs) can be used as a tool. hESCs are pluripotent with the abilities to be held in a self-renewal state or become induced to a desired germ layer then to be later directed through differentiation.
Rho Kinase Regulates Basement Membrane Dynamics To Coordinate Salivary Gland Branching Morphogenesis And Tissue Organization, William Patrick Daley
Rho Kinase Regulates Basement Membrane Dynamics To Coordinate Salivary Gland Branching Morphogenesis And Tissue Organization, William Patrick Daley
Legacy Theses & Dissertations (2009 - 2024)
Branching morphogenesis is a developmental mechanism utilized by many organs, including the salivary gland, lung, kidney, and mammary gland, to increase the epithelial surface area for secretion or absorption. The embryonic submandibular salivary gland (SMG) is a classic model for studying tissue morphogenesis in three dimensions ex vivo. Salivary gland development requires distinct but overlapping processes: morphogenesis, the actual physical rearrangement of cells into complex three dimensional structures, and cytodifferentiation, the process by which these cells begin to take on their own specialized function. Polarization, or the organization of cells into a cohesive tissue structure, also occurs concurrently during organ …
The P38 Alpha Stress-Activated Protein Kinase Pathway In Mammary Gland Development And Cancer, Huei-Chi Wen
The P38 Alpha Stress-Activated Protein Kinase Pathway In Mammary Gland Development And Cancer, Huei-Chi Wen
Legacy Theses & Dissertations (2009 - 2024)
Breast tumors arise from preinvasive lesions consisting of abnormal epithelial cells that accumulate within the mammary duct. However, the initiating steps and underlying pathways leading to the formation of these early stages of breast carcinoma, such as ductal carcinoma in situ (DCIS), is not fully understood.
Regulation Of The Aryl Hydrocarbon Receptor And The Signal Transducer And Activator Of Transcription 1 By Long-Term Estrogen Exposure In Breast Carcinoma Cells, Neal Englert
Legacy Theses & Dissertations (2009 - 2024)
The risk of developing breast cancer is known to be associated with a woman's lifetime exposure to estrogens, both endogenous and exogenous. Increased exposure to estrogens stimulates cellular proliferation, which is a widely accepted theory of estrogen receptor positive mammary carcinogenesis. The molecular mechanisms of gene expression regulation in response to long-term estrogen exposure (LTEE) of MCF-7 breast cancer cells were addressed in this study, with a focus on the aryl hydrocarbon receptor (AHR) and signal transducer and activator of transcription 1 (STAT1).
A Breast Cancer Stem Cell Model Created From Mmtv-Pymt Mice Applicable To Human Breast Cancer, Denise Grant Lanza
A Breast Cancer Stem Cell Model Created From Mmtv-Pymt Mice Applicable To Human Breast Cancer, Denise Grant Lanza
Legacy Theses & Dissertations (2009 - 2024)
Cancer stem cells are the seeds of tumor growth, but there are limited cell-based methods that exist to study the properties of these cells. To create a model of breast cancer stem cells, we isolated tumors from MMTV-PyMT mice. Two out of the four different cell types isolated survived in culture, CD44+CD24- and CD24+CD49f+CD44low. We found that we could initiate tumors with as few as 10 cells injected subcutaneously in the hind leg or orthotopically in the cleared fat pad with CD24+ cells. However, we could not initiate tumors with injection of CD24- cells. We found a requirement for TICs …
The Role Of Trm9 In Stress Responses, Ashish Ravindra Patil
The Role Of Trm9 In Stress Responses, Ashish Ravindra Patil
Legacy Theses & Dissertations (2009 - 2024)
Cells need to respond appropriately to environmental changes in order to maintain homeostasis. The cellular response to an environmental stress is regulated at transcriptional, translational and post translational levels. The tRNA, which acts as an adaptor molecule between the mRNA and the protein, plays an important role in the translational regulation of cellular responses to stress and is one of the most heavily modified biomolecules. In Saccharomyces cerevisiae , the wobble uracil of the tRNA(3'-UCU-5') Arg, tRNA(3'-UUC-5') Glu and certain other specific tRNAs are modified to 5-methoxycarbonylmethyluridine (mcm5U) and 5-methoxycarbonylmethyl-2-thiouridine (mcm5s2U) residues by the tRNA methyltransferase 9 (Trm9). Modifications at …
Live-Cell Studies On Mitotic Slippage In Humans, Daniela A. Brito
Live-Cell Studies On Mitotic Slippage In Humans, Daniela A. Brito
Legacy Theses & Dissertations (2009 - 2024)
Checkpoints are regulatory pathways that control the order and timing of specific cell-cycle events. In the presence of unattached/weakly-attached kinetochores, the mitotic checkpoint (MC) arrests cells in mitosis by inhibiting the degradation of cyclin B, the regulatory subunit of Cdk1 (cyclin dependent kinase 1). Checkpoints do not arrest cells permanently, and escaping mitosis with an unsatisfied MC requires cyclin B/Cdk1 inactivation. In yeast, this occurs through an “adaptation” mechanism involving inhibitory phosphorylations and/or Cdk1-inhibitors. To determine how vertebrate cells escape mitosis when the MC cannot be satisfied I conducted live-cell imaging and immunofluorescence studies on nocodazole-treated rat kangaroo (PtK) and …
An Rnai Screen Targeting The Protein Tyrosine Kinases Identifies Bruton's Tyrosine Kinase (Btk) As A Breast Cancer Cell Survival Factor, Cheryl Lynne Eifert
An Rnai Screen Targeting The Protein Tyrosine Kinases Identifies Bruton's Tyrosine Kinase (Btk) As A Breast Cancer Cell Survival Factor, Cheryl Lynne Eifert
Legacy Theses & Dissertations (2009 - 2024)
The receptor protein tyrosine kinases (RPTKs) and the non- receptor protein tyrosine kinases (PTKs) are among the most commonly up-regulated genes found in all types of cancers. Although, a large body of data implicates a majority of tyrosine kinases (TKs) in cancer, few have been extensively evaluated for any potential therapeutic benefit in any of the many subtypes of breast cancer. We have used RNA interference (RNAi) to perform a large-scale loss-of-function analysis to facilitate the identification of individual factors necessary for the survival of an ErbB2 positive breast cancer cell line. We have found that 30% of the TKs …