Cell and Developmental Biology Commons^™

Transposon-Mediated Stable Suppression Of Gene Expression In The Developing Chick Retina, Masaru Nakamoto, Chizu Nakamoto

Biology Faculty Publications

The embryonic chick has long been a favorite model system for in vivo studies of vertebrate development. However, a major technical limitation of the chick embryo has been the lack of efficient loss-of-function approaches for analyses of gene functions. Here, we describe a methodology in which a transgene encoding artificial microRNA sequences is introduced into embryonic chick retinal cells by in ovo electroporation and integrated into the genome using the Tol2 transposon system. We show that this methodology can induce potent and stable suppression of gene expression. This technique therefore provides a rapid and robust loss-of-function approach for studies of …

The Role Of Klf1 In Regulating Γ-Globin Gene Repressors, Anna P. Kovilakath

Theses and Dissertations

Sickle cell disease and β-thalassemia affect millions of people worldwide. γ-globin is the fetal counterpart to the adult β-globin. Research has shown that affected patients with higher than normal γ-globin show less severe symptoms. Therefore, reversing or preventing the hemoglobin switch from γ- to β- globin is a promising avenue of research for treating these diseases. KLF1 is an erythroid transcription factor involved in hemoglobin switching. Herein, we show that KLF1 directly regulates the γ-globin repressor gene LRF in both the mouse and human systems. KLF1 may also directly activate γ-globin expression by binding the promoter. In human HUDEP-2 cells, …

Vein Patterning During Juvenile Wing Development In Oncopeltus Fasciatus And Jadera Haematoloma, William R. Simmons

Honors Theses

Wings are a defining characteristic of all pterygote insects and are agreed to originate from a single common winged ancestor. However, essentially nothing is known about the molecular mechanisms that regulate wing development and patterning outside of Endopterygota, and the vast majority of this knowledge comes solely from Drosophila melanogaster. There is reason to suspect that the drastic developmental differences that exist between Endopterygota and Exopterygota have resulted in changes in the mechanisms, timings, or even genes that govern wing development. This study examined the roles of several genes known to be involved in D. melanogaster wing development and patterning …

Using Remote Site Incubators For Re-Introduction Of Arctic Graylng (Thymallus Arcticus) To The Big Manistee Watershed, Sunflower Wilson

Dissertations, Master's Theses and Master's Reports

Remote Site Incubators (RSIs) were developed for the incubation of salmonid eggs directly at a field site to enhance success and imprinting of young. These have been employed in the re-establishment of Arctic Grayling in Montana and are being laboratory tested for possible reintroduction of this species in the Big Manistee River, MI. Arctic Grayling, Rainbow and Brook Trout eggs, obtained from state hatcheries, were reared in a laboratory using flow through lake water to assess egg and fry survival using RSIs. Dead eggs and fry were removed daily and observations of developmental stages recorded. Rainbow Trout eggs were evaluated …

Investigating Mirna Functions In Arabidopsis By Short Tandem Target Mimic (Sttm), Yiyou Gu

Dissertations, Master's Theses and Master's Reports

MicroRNA (miRNAs) and other endogenous small RNAs play important role in regulating gene expression in eukaryotes. Till now, several techniques have been developed for miRNA function analysis. Because of the characteristic of miRNAs, the most applicable method for miRNA function research is blocking the whole miRNA family specifically. Here, we report an artificial non-coding transcript termed Short Tandem Target Mimic (STTM) for blocking miRNA activity in Arabidopsis thaliana. The STTM composed of two miRNA target sites with three nucleotides bulge in the miRNA cleavage site, the two-miRNA targets separated by spacer linker. We used this technique blocking several important endogenous …

Effects Of Chromium On Mouse Splenic T Lymphocytes And Effects Of Ethanol Exposure During Early Neurodevelopment On Behaviors In Mice, Lu Dai

Theses and Dissertations--Toxicology and Cancer Biology

The dissertation consists of three major projects with the focus on the immunotoxicity of chromium and the behavior disorders caused by early ETOH exposure respectively.

Hexavalent chromium [Cr(VI)] is widely used in various industrial processes and has been recognized as a carcinogen. As the first line of host defense system, the immune system can be a primary target of Cr(VI). T cell population represents a major arm of the immune system that plays a critical role in host anti-tumor immunity. Dysfunction of T cells compromises host anti-tumor immunity resulting in oncogenesis. Using mouse splenic T cells as an in vitro …

Alá Es Primero: Faith And Culture On The Mexican-American Border, Alan Ventura

Open Access Theses & Dissertations

The purpose of this Thesis is to show how El Paso Latino Muslims create a new cultural language when they harmonize their Mexican cultural practices with Islam.

Mammary Extracellular Matrix Directs Differentiation Of Testicular And Embryonic Stem Cells To Form Functional Mammary Glands In Vivo, Robert D. Bruno, Jodie M. Fleming, Andrea L. George, Corinne A. Boulanger, Pepper Schedin, Gilbert H. Smith

School of Medical Diagnostics & Translational Sciences Publications

Previously, we demonstrated the ability of the normal mammary microenvironment (niche) to direct non-mammary cells including testicular and embryonic stem cells (ESCs) to adopt a mammary epithelial cell (MEC) fate. These studies relied upon the interaction of transplanted normal MECs with non-mammary cells within the mammary fat-pads of recipient mice that had their endogenous epithelium removed. Here, we tested whether acellular mammary extracellular matrix (mECM) preparations are sufficient to direct differentiation of testicular-derived cells and ESCs to form functional mammary epithelial trees in vivo. We found that mECMs isolated from adult mice and rats were sufficient to redirect testicular derived …

Reproductive Competency And Mitochondrial Variation In Aged Syrian Hamster Oocytes, Fang Li, Frank J. Castora, Wentia Ford, Khalid Alarid, Howard W. Jones Jr., R. James Swanson

Biological Sciences Faculty Publications

The hamster is a useful model of human reproductive biology because its oocytes are similar to those in humans in terms of size and structural stability. In the present study we evaluated fecundity rate, ovarian follicular numbers, ova production, mitochondrial number, structure and function, and cytoplasmic lamellae (CL) in young (2–4 months) and old (12–18 months) Syrian hamsters (Mesocricetus auratus). Young hamsters had higher fertilisation rates and larger litters than old hamsters (100 vs 50% and 9.3 +/- 0.6 vs 5.5 +/- 0.6, respectively). Ovarian tissue from superovulated animals showed a 46% decrease in preantral follicles in old …

Ca2+/Calmodulin-Dependent Protein Kinase Type Ii (Camk-Ii) Is Required For Hematopoietic Stem Cell Specification, Camden E. Kurtz

Theses and Dissertations

Ca2+/Calmodulin-dependent protein kinase type II (CaMK-II) is a Serine/Threonine protein kinase that is activated by Ca2+ and Calmodulin to phosphorylate substrates involved in myriad developmental processes. This project implicates CaMK-II in specification of HSCs, and zebrafish provide an ideal embryonic model to study hematopoiesis. Zebrafish genetic manipulation was achieved through: incubation in chemical inhibitors; injection of notochord-targed WT and DN CaMK-II constructs with Transposase; and injection of camk2g1 translation-blocking morpholino antisense oligonucleotide (MO). Whole-mount in situ hybridization (WISH) and immunolocalization on zebrafish embryos allowed visualization of key HSC markers and pathway components that implicated CaMK-II in the specification of HSCs. …

Zebrafish (Danio Rerio) As A Model For Orofacial Research, Kevin A. Ghaffari

Theses and Dissertations

Across species, the face and more specifically the mouth, serves as an essential facet of everyday life. Amongst humans the mouth serves as a tool for the ingestion of food, a marker for facial recognition and a medium for communication. In order for the mouth to properly form, a series of precise growth and fusion events are needed. In order to insure that these events are orchestrated properly is a wide array of signals, transcription factors and epigenetic regulators. Due to the needed precision of these events, congenital birth defects of the face such as cleft lip and cleft palate …

Valproic Acid: A Neural Outgrowth Model For The Autism Spectrum Disorder, Hannah M. Bergan

Dissertations and Theses @ UNI

Autism spectrum disorder has been dramatically on the rise since the early 1990s, with one in forty-five children being diagnosed in 2014. It is known that contributing factors to autism spectrum disorder may have genetic and epigenetic origins, as well as environmental triggers. What is still unclear, however, is the impact of teratogenic drugs, consumed by the mother, have on a developing fetus in utero. How can they lead to the child developing autistic-like behaviors? The current study documents the effects of one teratogenic drug Valproic acid, a known histone deacetylase inhibitor. It establishes the role it plays on developing …

Sirt1 Regulates Glial Progenitor Proliferation And Regeneration In White Matter After Neonatal Brain Injury., Beata Jablonska, Marcin Gierdalski, Li-Jin Chew, Teresa Hawley, Mackenzie Catron, Arturo Lichauco, Juan Cabrera-Luque, Tracy Yuen, David Rowitch, Vittorio Gallo

Pediatrics Faculty Publications

Regenerative processes in brain pathologies require the production of distinct neural cell populations from endogenous progenitor cells. We have previously demonstrated that oligodendrocyte progenitor cell (OPC) proliferation is crucial for oligodendrocyte (OL) regeneration in a mouse model of neonatal hypoxia (HX) that reproduces diffuse white matter injury (DWMI) of premature infants. Here we identify the histone deacetylase Sirt1 as a Cdk2 regulator in OPC proliferation and response to HX. HX enhances Sirt1 and Sirt1/Cdk2 complex formation through HIF1α activation. Sirt1 deacetylates retinoblastoma (Rb) in the Rb/E2F1 complex, leading to dissociation of E2F1 and enhanced OPC proliferation. Sirt1 knockdown in culture …

Mathematical Analysis Of Feedback Targets Of Bmp Signaling In Drosophila Embryonic Development, Yan Luo

Open Access Theses

Bone morphogenetic proteins (BMPs) drive a range of cellular processes especially in the early stages of embryonic development. This family of proteins acts as one of the most important extracellular signals in development pattern formation across the animal kingdom. Cells in embryos differentiate into different cell types in response to the concentration level of BMP. This complex process is regulated by multiple regulators that serve to tune the signal response.

Extensive experimental and computational research has been performed to analyze BMP regulation in Drosophila, a widely studied model organism, and has advanced our understanding of animal development. Because of …

Nicotinamide Adenine Dinucleotide Phosphate Oxidase 2 Regulates Lps-Induced Inflammation And Alveolar Remodeling In The Developing Lung., Heather Menden, Sheng Xia, Sherry M. Mabry, Angels Navarro, Michael F. Nyp, Venkatesh Sampath

Manuscripts, Articles, Book Chapters and Other Papers

In premature infants, sepsis is associated with alveolar simplification manifesting as bronchopulmonary dysplasia. The redox-dependent mechanisms underlying sepsis-induced inflammation and alveolar remodeling in the immature lung remain unclear. We developed a neonatal mouse model of sepsis-induced lung injury to investigate whether nicotinamide adenine dinucleotide phosphate oxidase 2 (NOX2) regulates Toll-like receptor (TLR)-mediated inflammation and alveolar remodeling. Six-day-old NOX2

Retinoic Acid Signaling Regulates Krt5 Independently Of Stem Cell Markers In Submandibular Salivary Gland Epithelium., Timur Maratovich Abashev

Electronic Theses and Dissertations

Vitamin A metabolism, which produces the signaling molecule Retinoic Acid (RA), has been demonstrated to be important for growth and branching morphogenesis of mammalian embryonic salivary gland epithelium. However, it is not known whether RA functions directly within epithelial cells or in associated tissues that influence morphogenesis of salivary epithelium. Moreover, downstream targets of RA transcriptional regulation have not been identified. Here we show that canonical RA signaling occurs in multiple tissues of embryonic mouse salivary glands, including epithelium, associated parasympathetic ganglion neurons, and non‑neuronal mesenchyme. By culturing epithelium explants in isolation from other tissues we demonstrate that RA influences …

Effect Of Oxygen-Limiting Tidal Conditions On Muscle Metabolism And Structure In The Giant Acorn Barnacle, Balanus Nubilus, Katie O. Grady

Master's Theses

Crustacean muscle fibers are some of the largest cells in the animal kingdom, with fiber diameters in the giant acorn barnacle (Balanus nubilus) exceeding 3 mm. Sessile animals with extreme muscle sizes and that live in the hypoxia-inducing intertidal zone – like B. nubilus – represent ideal models for probing the effects of oxygen limitation on muscle cells. We investigated changes in metabolism and structure of B. nubilus muscle in response to: normoxic immersion, anoxic immersion, or air emersion, for acute (6h) or chronic (6h exposures twice daily for 2wks) time periods. Following exposure, we immediately measured hemolymph …

The Effects Of Sulfuric Acid Deposition On The Growth And Development Of Pond Breeding Salamanders In The Genus Ambystoma, Kenneth J. Anderson

Masters Theses & Specialist Projects

In terrestrial habitats with a history of mining activity and previous or ongoing reclamation efforts, understanding the effects of acidification on the ecology of amphibians is an important part of the restoration process and the conservation of local amphibian populations. Pond-breeding amphibians spend much of their post metamorphic life history in direct contact with the soil in upland habitat adjacent to aquatic breeding sites. I reared recently metamorphosed marbled salamanders (Ambystoma opacum) to evaluate the role of soil acidity on determinants of fitness such as growth and survival. My results indicate that a substrate of pH 4 was lethal to …

Multiple Sensory Modalities Used By Squid In Successful Predator Evasion Throughout Ontogeny, Carly A. York, Ian K. Bartol, Paul S. Kruger

Biological Sciences Faculty Publications

Squid rely on multiple sensory systems for predator detection. In this study we examine the role of two sensory systems, the lateral line analogue and vision, in successful predator evasion throughout ontogeny. Squid Doryteuthis pealeii and Lolliguncula brevis were recorded using high-speed videography in the presence of natural predators under light and dark conditions with their lateral line analogue intact or ablated via a pharmacological technique. Paralarval squid showed reduced escape responses when ablated; however, no differences were found between light and dark conditions in non-ablated paralarvae, as was previously shown in juveniles and adults, indicating that the lateral line …

Insulin-Like Growth Factor Pathway Described In Austrofundulus Limnaeus Diapause And Escape Embryos, Steven Cody Woll

Dissertations and Theses

Development in the annual killifish Austrofundulus limnaeus can follow two distinct developmental trajectories. Typical development includes the entrance of embryos into a state of metabolic and developmental arrest termed diapause. Alternately, embryos can escape diapause and develop directly without pause. These two trajectories are characterized by differences in the rate and timing of developmental, morphological, and physiological traits. Insulin and Insulin-like growth factor (IGF) signaling (IIS) is known to regulate entrance into diapause in a variety of invertebrates. In this thesis I explore the possible role of IGFs in the regulation of development and diapause in embryos of A. limnaeus …

Modeling And Analysis Of Germ Layer Formations Using Finite Dynamical Systems, Alexander Garza, Megan Eberle, Eric A. Eager

Spora: A Journal of Biomathematics

The development of an embryo from a fertilised egg to a multicellular organism proceeds through numerous steps, with the formation of the three germ layers (endoderm, mesoderm, ectoderm) being one of the first. In this paper we study the mesendoderm (the tissue that collectively gives rise to both mesoderm and endoderm) gene regulatory network for two species, \textit{Xenopus laevis} and the axolotl (\textit{Ambystoma mexicanum}) using Boolean networks. We find that previously-established bistability found in these networks can be reproduced using this Boolean framework, provided that some assumptions used in previously-published differential equations models are relaxed. We conclude by discussing our …

Investigating The Roles Of Δnp63 As A Suppressor Of Migration, Invasion, And Metastasis, Ramon E. Flores Gonzalez

Dissertations and Theses (Open Access)

Cancer is one of the leading causes of death and disease in the world. Considerable resources are spent to study and understand cancer, with the hope of developing new treatments and eventually cures that will help millions of people. Efforts to understand cancer are hindered by its inherent complexity and instability. Nonetheless, understanding the basics of tumor development and progression are the key to focused on studying the role of ΔNp63 in cancer, a p53 family member known to be involved in epithelial development, microRNA biogenesis, and stem cell maintenance. Using the strength of in vivo mouse models, we found …

Dichotomous Key To Pea Aphid (Acyrthosiphon Pisum) Apterous Parthenogenic Instars, Bates College Department Of Biology, Daisy Diamond, Daniel Levitis

SCARAB Data Repository

We provide a dichotomous key, with photographs and illustrations, for distinguishing between instars of the pea aphid (Acyrthosiphon pisum) in the developmental pathway leading to the apterous parthenogenetic adult. Lengths of body, antenna and cauda are provided for a sample of each instar.

Microrna-Target Interactions In Embryonic Stem Cells, Shuang Wu

Graduate School of Biomedical Sciences Theses and Dissertations

Animal embryogenesis is a highly orchestrated process involving many geneexpression programs that determine specific fates of individual cells during development. Gene expression can be regulated at different stages such as at transcription initiation or mRNA stability, and microRNA-mediated gene regulation is a crucial pathway on the regulation of mRNA stability. microRNAs, or miRNAs, are small, conserved non-coding RNAs that negatively regulate gene expression post-transcriptionally. The mouse miR-290-295 cluster and its human homolog miR-371-373 are stem cell-specific miRNA clusters, and their expression can only be observed in early embryos, primordial germ cells, germ cell compartments of adult testis and embryonic stem …

Genetic Analysis Reveals A Hierarchy Of Interactions Between Polycystin-Encoding Genes And Genes Controlling Cilia Function During Left-Right Determination, Daniel T. Grimes, Jennifer L. Keynton, Maria T. Buenavista, Xingjian Jin, Saloni H. Patel, Shinohara Kyosuke, Jennifer Vibert, Debbie J. Williams, Hiroshi Hamada, Rohana Hussain, Surya M. Nauli, Dominic P. Norris

Pharmacy Faculty Articles and Research

During mammalian development, left-right (L-R) asymmetry is established by a cilia-driven leftward fluid flow within a midline embryonic cavity called the node. This ‘nodal flow’ is detected by peripherally-located crown cells that each assemble a primary cilium which contain the putative Ca²⁺ channel PKD2. The interaction of flow and crown cell cilia promotes left side-specific expression of Nodal in the lateral plate mesoderm (LPM). Whilst the PKD2-interacting protein PKD1L1 has also been implicated in L-R patterning, the underlying mechanism by which flow is detected and the genetic relationship between Polycystin function and asymmetric gene expression remains unknown. Here, we …

The Role Of Daf-19 In Non-Ciliated Neurons: How Is Neural Development Regulated By Different Daf-19 Isoforms?, Zabdiel Ek Vazquez

Lawrence University Honors Projects

A degenerative disease-like phenotype, specifically reduction in synaptic protein levels in adult worms, is correlated with loss-of-function of the only RFX transcription factor gene, daf-19, in C. elegans. This gene encodes four known transcription factor isoforms, two of which are correlated with particular functions. The DAF-19C isoform activates genes responsible for cilia development, while DAF-19M is needed for cilia specification in males. A comparison of the transcriptome of daf-19 null and isogenic wild type adult worms suggests both positive and negative regulation of gene expression is correlated with the presence of DAF-19 proteins. We have assessed DAF-19 regulation …

Modelling The Polarization, Migration And Neuromast Deposition In The Zebrafish Posterior Lateral Line System, Hildur Knutsdottir

Biology and Medicine Through Mathematics Conference

No abstract provided.

Modeling Tracheal Occlusion In The Embryonic Lung, Uduak George

Biology and Medicine Through Mathematics Conference

No abstract provided.

Mechanics Of Early Retina And Lens Development In The Embryo, Alina Oltean

McKelvey School of Engineering Graduate Student Theses & Dissertations

Mechanical forces play an essential role in morphogenesis, the shaping of embryonic structures. This research focuses mainly on eye development, a problem that has been studied for decades using a variety of approaches. However, the mechanics of the early stages of eye formation remain incompletely understood.

The embryonic eyes begin as bilateral protrusions called optic vesicles (OVs) that grow outward from the anterior end of the brain tube. The optic vesicles contact and adhere to the overlying surface ectoderm (SE) via extracellular matrix (ECM). Then, both layers thicken in the region of contact to form the retinal and lens placodes, …

Embryonic And Post-Embryonic Hoxa13 Expression In The Four-Toed Salamander, Hemidactylium Scutatum., Breanna R. Lee

Senior Honors Projects, 2010-2019

The process of somitogenesis occurs during embryological development and results in the definition of persisting axial segments. The four toed salamander, Hemidactylium scutatum, exhibits post-embryonic segmentation while most other vertebrates stop segmentation at the end of embryological development. HoxA13, a transcription factor expressed along the developing anteroposterior axis, plays a role in specifying caudal segmental identity, cell ingression into the pre-somitic mesoderm (PSM), and PSM truncation. HoxA13’s responsibilities in influencing embryological development make it an interesting candidate for involvement in post-embryonic segment addition mechanisms. This study explores the role of HoxA13 in tail segmentation in H. scutatum embryonic, larval, …