Cell Biology Commons^™

Structure-Function Analysis Of Nel, A Thrombospondin-1-Like Glycoprotein Involved In Neural Development And Functions, Masaru Nakamoto

Biology Faculty Publications

Nel (neural epidermal growth factor (EGF)-like molecule) is a multimeric, multimodular extracellular glycoprotein with heparin-binding activity and structural similarities to thrombospondin-1. Nel is predominantly expressed in the nervous system and has been implicated in neuronal proliferation and differentiation, retinal axon guidance, synaptic functions, and spatial learning. The Nel protein contains an N-terminal thrombospondin-1 (TSP-N) domain, five cysteine-rich domains, and six EGF-like domains. However, little is known about the functions of specific domains of the Nel protein. In this study, we have performed structure-function analysis of Nel, by using a series of expression constructs for different regions of the Nel protein. …

Hyperosmotic Stress Enzyme Signaling Modulates Oct4, Nanog, And Rex1 Expression And Induces Prioritized Differentiation Of Murine Embryonic Stem Cells, Jill A. Slater

Wayne State University Dissertations

HYPEROSMOTIC STRESS ENZYME SIGNALING MODULATES OCT4, NANOG,

AND REX1 EXPRESSION AND INDUCES PRIORITIZED DIFFERENTIATION OF

MURINE EMBRYONIC STEM CELLS

by

JILL SLATER

MAY 2013

Advisor: Daniel Rappolee, Ph.D.

Major: Physiology

Degree: Doctor of Philosophy

Transcription factor expression and therefore lineage identity in the periimplantation

embryo and its stem cells may be influenced by extracellular stresses,

potentially affecting pregnancy outcome. Cellular stress forces cells to suppress some

normal activities (such as protein synthesis and cell proliferation) in order to repair

stress-damaged macromolecules and restore homeostasis. Therefore, any new

activities that embryonic cells initiate while concurrently funding the demands of the

stress …

A Proposal To Test The Effects Of Factor Ecat1 On Pluripotency, From Reprogramming To Differentiation Of Human Somatic Cells, Vritti R. Goel

CMC Senior Theses

The field of stem cell research has been growing more because of the interest in using stem cells to cure diseases and heal injuries. Human embryonic stem cells, because of the controversy surrounding them—and subsequently the difficulties in acquiring samples of the existing aging cell lines—can only be used in limited capacities. While the development of induced pluripotent stem cells in the last decade has allowed the field to progress closer to medical treatments, the low efficiency of reprogramming a somatic cell to a pluripotent state, and the vast molecular and genomic differences between human embryonic stem cells and human …

Interspecies Comparison Of Αii-Spectrin Abundance Between Chinook Salmon And Steelhead, Brielle D. Kemis, Ann L. Miracle, Katie A. Wagner, Christa M. Woodley

STAR Program Research Presentations

Salmonids, such as Chinook salmon (Oncorhynchus tshawytscha) and steelhead (O. mykiss), are a staple economic, recreational, tribal, and environmental resource, yet many populations are unsustainable. This study was part of a broad scale effort to monitor the impact of downstream migration obstacles on juvenile salmonid health and survival, which is an essential step towards increasing Smolt-to-Adult Return ratios (SARs). The objective of this study was to determine if juvenile Chinook salmon and steelhead exhibit differing quantities of alphaII-Spectrin Breakdown Products (SBDPs) over two consecutive spring migration periods, indicative of neurogenesis rate and/or biological response to head …

Enforced Expression Of Tbx1 In Fetal Thymic Epithelial Cells Antagonizes Thymus Organogenesis, Kim T. Cardenas

Dissertations and Theses (Open Access)

Enforced expression of Tbx1 in fetal thymic epithelial cells antagonizes

thymus organogenesis

Kim T. Cardenas

The thymus and parathyroid glands originate from organ-specific domains of 3rd pharyngeal pouch (PP) endoderm. At embryonic day 11.5 (E11.5), the ventral thymus and dorsal parathyroid domains can be identified by Foxn1 and Gcm2 expression respectively. Neural crest cells, (NCCs) play a role in regulating patterning of 3rd PP endoderm. In addition, pharyngeal endoderm influences fate determination via secretion of Sonic hedgehog (Shh), a morphogen required for Gcm2 expression and generation of the parathyroid domain. Gcm2 is a downstream target of the transcription factor Tbx1, …

Analysis Of Band 4.1b In Integrin-Mediated Cell Adhesion And Signaling, Youngsin Jung

Dissertations and Theses (Open Access)

Band 4.1B is a cytoskeletal adaptor protein that regulates various cellular behavior; however, the mechanisms by which Band 4.1B contributes to intracellular signaling are unclear. This project addresses in vivo and in vitro functions for Band 4.1B in integrin-mediated cell adhesion and signaling. Band 4.1B has been shown to bind to β8 integrin, although cooperative functions of these two proteins have not been determined. Here, functional links between β8 integrin and Band 4.1B were investigated using gene knockout strategies. Ablation of β8 integrin and Band 4.1B genes resulted in impaired cardiac morphogenesis, leading to embryonic lethality by E11.5. These embryos …

Developmental And Cellular Functions Of Delta-Catenin, Dongmin Gu

Dissertations and Theses (Open Access)

Catenins have diverse and powerful roles in embryogenesis, homeostasis or disease progression, as best exemplified by the well-known beta-catenin. The less studied delta-catenin likewise contains a central Armadillo-domain. In common with other p120 sub-class members, it acts in a variety of intracellular compartments and modulates cadherin stability, small GTPase activities and gene transcription. In mammals, delta-catenin exhibits neural specific expression, with its knock-out in mice correspondingly producing cognitive defects and synaptic dysfunctions.

My work instead employed the amphibian, Xenopus laevis, to explore delta-catenin’s physiological functions in a distinct vertebrate system. Initial isolation and characterization indicated delta-catenin’s expression in Xenopus. Unlike …

The Role And Mechanism Of The Homeobox Gene Dlx4 In Transforming Growth Factor-B Resistance In Cancer, Bon Q. Trinh

Dissertations and Theses (Open Access)

Transforming growth factor-b (TGF-b) is a cytokine that plays essential roles in regulating embryonic development and tissue homeostasis. In normal cells, TGF-b exerts an anti-proliferative effect. TGF-b inhibits cell growth by controlling a cytostatic program that includes activation of the cyclin-dependent kinase inhibitors p15^Ink4B and p21^WAF1/Cip1 and repression of c-myc. In contrast to normal cells, many tumors are resistant to the anti-proliferative effect of TGF-b. In several types of tumors, particularly those of gastrointestinal origin, resistance to the anti-proliferative effect of TGF-b has been attributed to TGF-b receptor or Smad mutations. However, these mutations are absent from many …

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

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.

Defining The Role Of Il-15 Trans-Presentation By Distinct Cell-Types During The Development And Homeostasis Of Natural Killer And Invariant Natural Killer T Cells, Eliseo F. Castillo

Dissertations and Theses (Open Access)

The immuno-regulatory functions displayed by NK and iNKT cells have highlighted their importance as key lymphocytes involved in innate and adaptive immunity. Therefore, understanding the dynamics influencing the generation of NK and iNKT cells is extremely important. IL-15 has been shown to provide a critical signal throughout the development and homeostasis of NK and iNKT cells; however, the cellular source of IL-15 has remained unclear. In this investigation, I provide evidence that the cell-type providing IL-15 to NK and iNKT cells via trans-presentation is determined by the tissue site and the maturation status of NK and iNKT cells. For NK …

Determination Of The Myogenic Potential Of Human Embryonic Stem Cell-Derived Mesenchymal Stem Cells, Rory Coleman

Honors Scholar Theses

Human embryonic stem cells (hESCs) have the potential to

differentiate to all adult somatic cells. This property makes hESCs a very promising area of research for the treatment of disorders in which specific cell populations need to be restored. Despite this potential, research that focuses on producing mesodermally derived cell populations from hESCs is decidedly limited, notwithstanding the prevalence of disorders involving mesodermal tissues for which treatment options are limited. Skeletal muscle myoblasts are derivatives of mesodermal cells and are characterized by the expression of the MyoD gene. These cells are difficult to obtain from hESCs in a reproducible and …

Validation Of The Activation Of Aurora B Kinase By Caenorhabditis Elegans To Usled-Like Kinase And The Identification Of Cyclin B3 As A Phospho-Specific Tlk-1 Interactor, Gary Michael Deyter

Dissertations and Theses (Open Access)

A hallmark of tumorigenesis and certain birth defect syndromes is the loss of ploidy that can result from incorrect chromosome segregation. Chromosomes that are not partitioned properly during mitosis are often fragmented, changing the genetic makeup of daughter cells. Inheriting extrachromosomal fragments that contain cell survival genes or losing chromosomal loci that encode tumor suppressors can promote tumor development. Thus, it is essential to elucidate molecular mechanisms required for correct chromosome segregation. Chromosomes are connected to mitotic spindle microtubules by way of a proteinacous, chromosome-bound organelle called the kinetochore. Two decades of research have confirmed that the conserved Aurora B/AIR-2 …

The Consequences Of Disrupting The Mdm2-P53 Balance In Hematopoiesis, Hussein A. Abbas

Dissertations and Theses (Open Access)

The bone marrow accommodates hematopoietic stem cells and progenitors. These cells provide an indispensible resource for replenishing the blood constituents throughout an organism’s life. A tissue with such a high turn-over rate mandates intact cycling checkpoint and apoptotic pathways to avoid inappropriate cell proliferation and ultimately the development of leukemias. p53, a major tumor suppressor, is a transcription factor that regulates cell cycle, and induces apoptosis and senescence. Mice inheriting a hypomorphic p53 allele in the absence of Mdm2, a p53 inhibitor, have elevated p53 cell cycle activity and die by postnatal day 13 due to hematopoietic failure. Hematopoiesis progresses …

Cellular And Developmental Functions Of The Xenopus Arvcf-Catenin:Kazrin Complex, Kyucheol Cho

Dissertations and Theses (Open Access)

Xenopus ARVCF (xARVCF), a member of p120-catenin subfamily, binds cadherin cytoplasmic domains to enhance cadherin metabolic stability, or when dissociated, modulates Rho-family GTPases. We previously found that xARVCF binds directly to Xenopus KazrinA (xKazrinA), a widely expressed, conserved protein that bears little homology to established protein families. xKazrinA is also known to influence keratinocyte proliferation-differentiation and cytoskeletal activity. In my study, I first evaluated the expression pattern of endogenous Kazrin RNA and protein in Xenopus embryogenesis as well as in adult tissues. We then collaboratively predicted the helical structure of Kazrin’s coiled-coil domain, and I obtained evidence of Kazrin’s dimerization/oligomerization. …

Phylogeography Of The Frog Leptodactylus Validus (Amphibia: Anura): Patterns And Timing Of Colonization Events In The Lesser Antilles, Arley Camargo, W. Ronald Heyer, Rafael O. De Sá

Biology Faculty Publications

The frog Leptodactylus validus occurs in northern South America, Trinidad and Tobago, and the southern Lesser Antilles (Grenada and St. Vincent). Mitochondrial DNA sequences were used to perform a nested clade phylogeographic analysis (NCPA), to date colonization events, and to analyze colonization patterns using on a relaxed molecular clock and coalescent simulations. L. validus originated on the mainland and first colonized Trinidad with subsequent independent colonizations of Tobago and the Lesser Antilles from Trinidad. The NCPA suggests a historical vicariant event between populations in Trinidad and Tobago from those in the Lesser Antilles. The colonization of Trinidad occurred 1 million …

In Vivo Analysis Of The Notch Receptor S1 Cleavage, Robert J. Lake, Lisa M. Grimm, Alexey Veraksa, Spyros Artavanis-Tsakonas

Biology Faculty Publication Series

A ligand-independent cleavage (S1) in the extracellular domain of the mammalian Notch receptor results in what is considered to be the canonical heterodimeric form of Notch on the cell surface. The in vivo consequences and significance of this cleavage on Drosophila Notch signaling remain unclear and contradictory. We determined the cleavage site in Drosophila and examined its in vivo function by a transgenic analysis of receptors that cannot be cleaved. Our results demonstrate a correlation between loss of cleavage and loss of in vivo function of the Notch receptor, supporting the notion that S1 cleavage is an in vivo mechanism …

Bmp Signaling Goes Posttranscriptional In A Microrna Sort Of Way, Catherine A. Reinke, Richard W. Carthew

Faculty Publications

Aberrant microRNA (miRNA) expression correlates with human diseases such as cardiac disorders and cancer. Treatment of such disorders using miRNA-targeted therapeutics requires a thorough understanding of miRNA regulation in vivo. A recent paper in Nature by Davis et al. expands our understanding of miRNA biogenesis and maturation, elucidating a mechanism by which extracellular signaling directs cell differentiation via posttranscriptional regulation of miRNA expression.

Is The Amphibian Tree Of Life Really Fatally Flawed?, Darrel R. Frost, Taran Grant, Julian Faivovich, Raoul H. Bain, Alexander Haas, Celio F.B. Haddad, Rafael O. De Sá, Et Al.

Biology Faculty Publications

Wiens (2007, Q. Rev. Biol. 82, 55–56) recently published a severe critique of Frost et al.'s (2006, Bull. Am. Mus. Nat. Hist. 297, 1–370) monographic study of amphibian systematics, concluding that it is “a disaster” and recommending that readers “simply ignore this study”. Beyond the hyperbole, Wiens raised four general objections that he regarded as “fatal flaws”: (1) the sampling design was insufficient for the generic changes made and taxonomic changes were made without including all type species; (2) the nuclear gene most commonly used in amphibian phylogenetics, RAG-1, was not included, nor were the morphological characters that had justified …

Phylogenetic Analyses Of Mtdna Sequences Reveal Three Cryptic Lineages In The Widespread Neotropical Frog Leptodactylus Fuscus (Schneider, 1799) (Anura, Leptodactylidae), Arley Camargo, Rafael O. De Sá, W. Ronald Heyer

Biology Faculty Publications

Leptodactylus fuscus is a neotropical frog ranging from Panamá to Argentina, to the east of the Andes mountains, and also inhabiting Margarita, Trinidad, and the Tobago islands. We performed phylogenetic analyses of 12S rRNA, 16S rRNA, tRNA-Leu, and ND1 mitochondrial (mt) DNA sequences from specimens collected across the geographic distribution of L. fuscus to examine two alternative hypotheses: (i) L. fuscus is a single, widely distributed species, or (ii) L. fuscus is a species complex. We tested statistically for geographic association and partitioning of genetic variation among mtDNA clades. The mtDNA data supported the hypothesis of several cryptic species within …

Skeletal Myosin Heavy Chain Function In Cultured Lung Myofibroblasts, Nancy A. Rice, Leslie A. Leinwand

Biology Faculty Publications

Myofibroblasts are unique contractile cells with both muscle and nonmuscle properties. Typically myofibroblasts are identified by the expression of smooth muscle actin (ASMA); however some myofibroblasts also express sarcomeric proteins. In this study, we show that pulmonary myofibroblasts express three of the eight known sarcomeric myosin heavy chains (MyHCs) (IIa, IId, and embryonic) and that skeletal muscle myosin enzymatic activity is required for pulmonary myofibroblast contractility. Furthermore, inhibition of skeletal myosin activity and myofibroblast contraction results in a decrease in both ASMA and skeletal MyHC promoter activity and ASMA protein expression, suggesting a potential coupling of skeletal myosin activity and …

The Wings Of Bombyx Mori Develop From Larval Discs Exhibiting An Early Differentiated State: A Preliminary Report, Madhuri Kango-Singh, Amit Singh, K. P. Gopinathan

Biology Faculty Publications

Lepidopteran insects present a complex organization of appendages which develop by various mechanisms. In the mulberry silkworm, Bombyx mori, a pair of meso- and meta-thoracic discs located on either side in the larvae gives rise to the corresponding fore- and hind-wings of the adult. These discs do not experience massive cell rearrangements during metamorphosis and display the adult wing vein pattern. We have analysed wing development in B. mori by two approaches, viz., expression of patterning genes in larval wing discs, and regulatory capacities of larval discs following explantation or perturbation. Expression of Nubbin is seen all over the presumptive …

Chondrocranium And Visceral Skeleton Of Atelopus Tricolor And Atelophryniscus Chrysophorus, E O. Lavilla, Rafael O. De Sá

Biology Faculty Publications

Atelopus tricolor and Atelophryniscus chrysophorus have gastromyzophorous tadpoles. Aside from specific differences, the larval chondrocrania and visceral skeletons show several shared character states, including an almost quadrangular, open chondrocranium , cornua trabeculae with expanded tips, cartilago suprarostralis as a single element, a posteriorly projected arcus subocularis quadrati, a closed muscular tunnel, and a simplified branchial basket, with only three pairs of ceratobranchiales. They are compared with diverse character states of suctorial larvae considering them as causally related to the stream life adaptations.

Do Timing And Pattern Of Myogenesis Correlate With Life History Mode In Anurans?, Matthew T. Smetanick, Rafael O. De Sá, Gary P. Radice

Biology Faculty Publications

The timing and pattern of myogenesis varies among anurans that have been studied and the different pat- terns may provide useful phylogenetic information. Specific myogenic markers have been described (Muntz, 1975; Kielbowna, 1981; Boudjelida & Muntz, 1987; Radice et al., 1989) and they can provide infor- mation on evolutionary changes for closely related lineages within a clade. For example, we previously com- pared first appearance of a muscle-specific protein, first twitch of axial muscle, onset of multinucleation within axial myotome, and first heartbeat in two pipid genera (Smetanick et al., 1999). We found that although the timing of myogenesis differed, …

Development Of The Suprarostral Plate Of Pipoid Frogs, Rafael O. De Sá, Charles C. Swart

Biology Faculty Publications

The rostral region of nonpipoid tadpoles has two sets of cartilages, the cornua trabeculae and the suprarostral cartilages, whereas the rostral region in pipoid larvae is occupied by a single and continuous cartilage, the suprarostral plate. The homology of this region in pipoid and nonpipoids tadpoles has been controversial. We examined the early formation and development of the suprarostral plate using serially cross-sectioned specimens of Rhinophrynus, Xenopus, and Hymenochirus. We conclude that the cartilaginous structures present in the rostral area of pipoid and nonpipoid larvae are homologous. Furthermore, we found two different developmental patterns among pipoid larvae. The chondrocranium …

The Biology Of Xenopus By R. C. Tinsley And H. C. Kobel, Rafael O. De Sá

Biology Faculty Publications

The Biology of Xenopus presents a summary of current knowledge about a single genus resulting from a symposium held at the Zoological Society of London in September 1992. This approach to summarizing available information has also been taken for other taxa, such as Atelopus (Lotters, 1996). However, the task of compiling data for Xenopus is enormous relative to any other amphibian group, because Xenopus laevis has become a model system for molecular and development research (Cannatella and de Sa, 1993). Unfortunately, most of our knowledge of Xenopus is biased toward this single species. There are about 20 recognized species of …

Chondrocranial Morphology Of Leptodactylus Larvae (Leptodactylidae: Leptodactylinae): Its Utility In Phylogenetic Reconstruction, Peter M. Larson, Rafael O. De Sá

Biology Faculty Publications

Chondrocranial morphology of leptodactylid frogs is scarcely known and has not been completely described for any species of Leptodactylus. We describe the diversity of chondrocranial morphology in the genus Leptodactylus based on the analysis of 22 species, representing the four species groups: the fuscus Group, ocellatus Group, melanonotus Group, and pentadactylus Group. Furthermore, 26 characters are identified and used in a phylogenetic analysis. The phylogenetic analysis using Physalaemus, Crossodactylus, and Hylodes as outgroups suggests two monophyletic clades within Leptodactylus: the melanonotus-ocellatus clade and the pentadactylusfuscus clade. However, it does not support the monophyly of the species groups as currently recognized …

Phase I Trial Of Iodine-131-Chimeric B72. 3 (Human Igg4) In Metastatic Colorectal Cancer, Ruby F. Meredith, M B. Khazaeli, Walter Plott, Mansoor Saleh, Tiepu Liu, Laquetta Allen, Charles Russell, Roger Orr

Haematology and Oncology, East Africa

Twelve patients with metastatic colorectal cancer participated in a Phase I trial of 1311-labeled chimeric B72.3 (human IgG4). Consecutive groups of patients received 18 mCi/m 2, 27 mCi/ m 2 and 36 mCi/m a. No acute side effects related to antibody administration were noted. Bone marrow suppression was the only side effect; it was dose-dependent and correlated with whole-body radiation dose estimates. The lowest dose level produced no marrow suppression, whereas 27 mCi/m 2 resulted in Grade 1 and 2 marrow suppression in two of three patients. The maximum tolerated close was 36 mCi/m 2 with all six patients at …

Fertilization And Mouse Embryo Development In The Presence Of Midazolam, Maria Leavitt

Biological Sciences Theses & Dissertations

The development of mouse embryos in vitro has provided a unique opportunity for the study of cellular differentiation and the influence of various agents on differentiation. Midazolam is administered for conscious sedation in the recovery of human ova. In the Norfolk IVF Program, midazolam is found in varying concentrations in follicular fluid. Mouse embryo formation and development has been used to test midazolam's effect on fertilization and early development as a model of human in vitro fertilization and development. Midazolam' s effects were studied using varying doses of midazolam on (1) in vivo fertilization and subsequent cell division, and (2) …

Cell Surface-Binding Sites For Progesterone Mediate Calcium Uptake In Human Sperm, Peter F. Blackmore, Joseph Neulan, Frank Lattanzio, Stephen J. Beebe

Bioelectrics Publications

Recent studies (e.g. Blackmore, P. F., Beebe, S. J., Danforth, D. R., and Alexander, N.) (1990) J. Biol. Chem. 265, 1376-1380) have shown that in human sperm, progesterone produces a rapid increase in intracellular free calcium ([Ca2+]i) and an induction of the acrosome reaction (e.g. Osman, R. A., Andria, M. L., Jones, A. D., and Meizel, S. (1989) Biochem. Biophys. Res. Commun. 160, 828-833). In this study, the location of progesterone receptors on the cell surface of human sperm was identified using progesterone immobilized on bovine serum albumin (BSA) (progesterone 3-(O-carboxymethyl)oxime:BSA) as well as progesterone and its 3-O-carboxymethyloxime derivative. Using …