Developmental Biology Commons^™

Evaluation Of The Bax-Vdac Interaction And Their Influence On Apoptosis In Drosophila Melanogaster, Frances Marie Gatlin

Honors Theses

Apoptosis, also known as programmed cell death, is a cellular process used for development or for when cells undergo injury or stress. The Bcl2 family of proteins includes both pro-apoptotic and anti-apoptotic proteins that control the intrinsic pathway of apoptosis. Understanding the mechanisms and influence these proteins have on apoptosis is an important area of research focused on in Dr. Jones’s lab. Evidence shows a homology amongst the Bcl2 family of proteins at the BH3 domain. Dr. Jekabsons' lab has found a potential homology amongst VDAC 1-3 and the Bcl2 family at the BH3 domain.

Specifically, our lab is using …

Elucidating The Functional Location Of Platelet-Derived Growth Factor Receptor Alpha, Creation Of A Review Chapter On The Use Of Zebrafish In Studying Congenital Heart Defects, And Using 3d Printing To Create Laboratory Tools, Jaret Lieberth

Honors Theses

Congenital heart defects (CHDs) are currently the most prevalent form of birth defect in the United States. Their combined frequency and severity make necessary a deeper understanding of the molecular underpinnings guiding heart formation. The first major step in cardiac morphogenesis, cardiac fusion, involves the medial movement of bilateral populations of myocardial precursor cells to the embryonic midline, where they merge to form the primitive heart tube. Although crucial to subsequent organogenesis, the precise mechanisms governing cardiac fusion remain unknown. Previous studies have found that a mutation in platelet-derived growth factor receptor alpha (pdgfra), called refuse-to-fuse (ref), results …

Multi-Generational Effects Of ∆9-Tetrahydrocannabinol Exposure On Gene Expression In Liver Tissue, Kayla Lovitt

Honors Theses

Cannabis is the most commonly used, cultivated, and trafficked illicit drug worldwide. Increased availability and acceptance of cannabis and cannabinoid-containing products provide the necessity for understanding how these substances influence aging. In this study, zebrafish (Danio rerio) were exposed to concentrations of Δ⁹-tetrahydrocannabinol (THC) (0.08, 0.4, 2 µM) during embryonic-larval development, the effects on aging were measured 30 months later and in the offspring of the exposed fish (F1 generation. We observed results indicating a biphasic and hormetic effect. Treatment with the lowest concentration of THC significantly increased egg production, while higher concentrations resulted in impaired …

Microglia And Fractalkine Involvement In Shaping Multimodal Midbrain Circuitry, Cooper A. Brett

Masters Theses, 2020-current

Microglial cells (MGCs) are highly dynamic and have been implicated in shaping discrete neural maps in several systems. MGCs respond to numerous cues in their microenvironment, including a neuronally-expressed chemokine, CX3CL1 (fractalkine). The present study examines microglial and CX3CL1 patterns with regard to the emerging modular-extramodular framework within the lateral cortex of the inferior colliculus (LCIC). The LCIC is a shell region of the auditory midbrain where discrete compartments receive modality-specific connections, whereby somatosensory inputs terminate within modules and auditory inputs target surrounding extramodular zones. A reliable modular marker, glutamic acid decarboxylase (GAD), enables visualization of emerging modular domains in …

Using A Novel Cell- Penetrating Peptide Technology To Induce Pluripotency, Laura-Maria Oja

Master of Science in Chemical Sciences Theses

Somatic stem cells have been used as a regenerative medicine tool to treat diseases like leukemia for decades, but they are limited in their ability to self-renew and differentiate. A better alternative, induced pluripotent stem cells (iPSCs), has the ability to generate cells from all three germ layers and avoid tissue rejection as well as bypass the ethical concerns related to embryonic stem cells. In 2006, Takahashi and Yamanaka discovered that only four genes were required to induce pluripotency: Oct4, Sox2, Klf4, and c-Myc (OSKM). Since then, pluripotency has been induced with OSKM most effectively by transfection or transduction. However, …

Neurocognitive Risk Factors And Current Intervention Strategies For Survivors Of Pediatric Acute Lymphoblastic Leukemia, Abigail Taber

Senior Honors Theses

The improved survival rate for pediatric cancer patients is one of the greatest triumphs of recent medicine, but the late effects faced by these survivors have been uncovered through this new population of survivors. Many survivors of pediatric acute lymphoblastic leukemia (ALL) experience cognitive deficits in areas such as attention, memory, processing speed, and academic achievement following cancer treatment. Recent research has pointed to chemotherapeutic agents, host risk factors, and genetic predispositions as perpetrators of these deficits, although other factors are also under investigation. Consequently, the search for appropriate interventions for the amelioration of these deficits has dominated the literature …

Development Of A Crispr System To Mutate Lpar4 In Chick Retinal Ganglion Cells, Garrett Driscoll

Graduate Theses

Growth cones direct axon pathfinding during neural development by detecting environmental stimuli, known as axon guidance molecules. In vitro studies have shown inhibitory axon guidance molecules to cause growth cones to change morphology to what is called a collapsed growth cone. Lysophosphatidic acid (LPA) has been demonstrated to collapse growth cones in culture, and thus it may act as an inhibitory molecule. However, identification of what LPA receptor is responsible for this physiological response is unknown. To investigate which receptor elicits collapse, we focused on LPA receptor-4 (LPA4) by designing a CRISPR construct to mutate Lpar4 in chick retinal ganglion …

Microglia-Neuron Interactions In A Mouse Model Of Low Grade Neuroepithelial Tumors, Veolette Hanna

Honors Scholar Theses

Microglia are the macrophages of the brain and spinal cord, playing an important role in the immune response to disease states of the nervous system. This study conducts an investigation on the activity of microglia in response to low grade neuroepithelial tumors. Using mouse models and microglial markers, a qualitative and quantitative analysis of microglia activation, migration, and invasion within the brain cortex during early stages of tumor development was conducted. It was found that the presence of a low grade neuroepithelial tumor in the cortex of one hemisphere of the brain causes significant microglia activation in comparison to the …

Serpinb1 Effects On Human Pancreatic Ductal Cells, Praneeth Goli

College of Arts & Sciences Senior Theses

Rebuilding or expanding the body’s ability to regulate blood glucose levels by restoring or increasing the body’s ability to produce insulin, long-term, would be a major advancement in diabetes (type-1 and type-2) treatment and could serve as a potential cure [1]. Therefore, the holy grail of diabetes treatment and cure research is to regenerate or increase the insulin producing cells in diabetic individuals (type-1 and type-2) [1, 2]. SerpinB1 has been implicated as a biomolecular signal that stimulates an increase in insulin cell mass in murine and zebrafish models that exhibit physiologically normal compensatory islet hyperplasia. In humans, SerpinB1 is …

Mouse Embryonic Stem Cells Are Sensitive To The Cytotoxicity Of Nitric Oxide: Biological Implications For Early Embryogenesis, Mckenzie C. Hargis

Honors Theses

The early embryo, before implantation, is at a very vulnerable stage in development where it faces various inflammatory cytokines throughout the implantation process. In this stage, the cells in the blastocyst, the preimplantation stage embryo, must proliferate rapidly for tissue formation. However, it is known that inflammatory cytokines can inhibit cell proliferation. Previous studies have shown that embryonic stem cells (mESCs), the major cell component in the blastocyst, are unresponsive to treatments of tumor necrosis factor ⍺ (TNF⍺) and interferon 𝛾 (IFN𝛾), two inflammatory cytokines involved in the implantation process. Treatment of mESC-differentiated fibroblasts (mESC-FBs) with TNF⍺ and IFN𝛾 in …

Investigating Roles For Rna Turnover Processes In Cell Signaling Through Drosophila Melanogaster Genetic Mosaics, Sudiksha Rathan Kumar

Honors Theses

The process of cell signaling is vital in organisms for proper development as it determines the fate and function of cells. This process is highly regulated by myriad interactions between signaling pathway components and gene expression mechanisms. RNA turnover is a type of RNA processing that degrades RNA. It plays an essential role in homeostasis and environmental changes; however, its influence on signaling pathways is currently unknown. In the present study, the effect of RNA turnover processes on cell signaling was analyzed using the genetic mosaics in Drosophila melanogaster. Fly mosaics were created by breeding RNAi lines targeting cellular …

Elucidating The Developmental Defects In Zebrafish Associated With The Cardiac Drug Verapamil, Blake Stephan Justis

Graduate Theses/Dissertations

Birth defects are abnormalities in a developing organism that lead to a malformation in structure or function. Over half of birth defects have no determined cause; however, known causes occur by genetic anomalies, exposure to environmental agents (a.k.a. teratogens), or multifactorial reasons. To explain the unknown causes of birth defects, an area of focus in this study is to identify potential teratogens. Identifying these teratogens, is key to preventing future birth defects. An obvious source of teratogens in pregnant women would be that of pharmaceuticals. Thus, a main goal of this study is to identify drugs that cause birth defects. …

Otx2 Represses Sister Cell Fate Choices In The Developing Retina To Promote Photoreceptor Specification, Miruna Georgiana Ghinia Tegla, Diego F. Buenaventura, Diana Y. Kim, Cassandra Thakurdin, Kevin C. Gonzalez, Mark M. Emerson

Publications and Research

During vertebrate retinal development, subsets of progenitor cells generate progeny in a non-stochastic manner, suggesting that these decisions are tightly regulated. However, the gene-regulatory network components that are functionally important in these progenitor cells are largely unknown. Here we identify a functional role for the OTX2 transcription factor in this process. CRISPR/Cas9 gene editing was used to produce somatic mutations of OTX2 in the chick retina and identified similar phenotypes to those observed in human patients. Single cell RNA sequencing was used to determine the functional consequences OTX2 gene editing on the population of cells derived from OTX2-expressing retinal progenitor …

Cd11b+ Depletion Of Bone Marrow Derived Macrophages Negatively Affects Bone Fracture Healing, Sarah Hozain

Seton Hall University Dissertations and Theses (ETDs)

Hematoma formation and inflammation occurs during the beginning stages of fracture repair, which require the presence of innate cells such as macrophages. Macrophages are subdivided into proinflammatory, M1, or anti-inflammatory, M2, phenotypes with different functions, cytokine profiles, and surface markers. In this study, in vitro and in vivo models were used to deplete M1 macrophages, using Mac-1 Sap conjugated antibody (Mac1SAP), to determine the effects on fracture healing. In vitro study, primary macrophages isolated from mice femoral bone marrow were harvested and differentiated into M1 macrophages (+LPS), M2 macrophages (+IL-4), or undifferentiated then treated with either vehicle or 10pM Mac1SAP. …

Profiling Of Small Non-Coding Rna In Bovine Somatic Cell Nuclear Transfer Embryos During Early Development Through The Maternal-To-Embryonic Transition, Abby D. Benninghoff, Jocelyn Cuthbert

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Supplementary File 1. Read lengths and annotation summaries of sncRNA: Microsoft Excel document with information from small RNA sequencing on the aligned, annotated reads.

Supplementary File 2. Normalized and variance-stabilized values for annotated sncRNAs: Microsoft Excel document with normalized and variance-stabilized reads generated by RNA sequencing of small non-coding RNA from cattle embryos.

Supplementary File 3. Results of DESeq2 analyses for miRNAs, tRNA fragments and piRNAs: Microsoft Excel document with results of DESeq2 differential expression analysis comparing scNT vs IVF embryos at each developmental stage and sequential comparisons of developmental stage for both IVF and scNT embryos.

Supplementary File 4. …

Mapping Transcriptional Changes Associated With The Maternal-To-Embryonic Transition In Bovine Embryos To Small Non-Coding Rna Profiles In Both In Vitro-Fertilized And Scnt Cattle Embryos, Abby D. Benninghoff, Jocelyn Cuthbert

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Supplementary File 1. Normalized reads for annotated mRNA transcripts. Microsoft Excel document providing normalized sequencing reads per sample.

Supplementary File 2. Results of DESeq2 analyses for annotated mRNAs. Microsoft Excel document with results of DESeq2 differential expression analysis comparing scNT vs IVF embryos at each developmental stage and sequential comparisons of developmental stage for both IVF and scNT embryos.

Supplementary File 3. Results of Metascape ontology analyses for differentially expressed mRNAs. Microsoft Excel document with results of Metascape ontology analyses for biological processes associated with predicted mRNA targets of differentially expressed miRNAs.

Supplementary File 4. Cytoscape networks for differentially expressed …

155— Analysis Of Her4.1 And Ascl1a In Gef Mutants, Rico Amato, Tessa Beiter, Christopher Lepore

GREAT Day Posters

Zebrafish are a model organism for studying developmental abnormalities, especially in the eye. The good effort (gef) mutant zebrafish have smaller eyes than wild-type embryos due to rapid retinal degeneration that becomes apparent only after two days post fertilization. The genetic problem arises from a deletion of intronic DNA sequence which leads to the loss of an exon and disrupts the coding region of the Cha1b protein. Chaf1b is a subunit of the Chromosome assembly factor 1 (CAF-1), a complex of three proteins which has a role in histone loading and chromatin regulation. This small eye phenotype has been hypothesized …

Hers And His: Silk Glands Used In Egg Sac Construction By Female Spiders Potentially Repurposed By A 'Modern' Male Spider, Mark A. Townley, Danilo Harms

Biological Sciences

Cylindrical silk gland (CY) spigots distinguish a large clade of modern spiders, the CY spigot clade, which includes all entelegyne spiders and their closest relatives. Following a widespread paradigm, CYs and their spigots are only known to occur in female spiders and they produce silk used in the construction of egg sacs. Here we report the occurrence of a CY spigot or CY nubbin on each posterior median spinneret (PMS) in males (5th stadium and later) of the spider Australomimetus maculosus. Late juvenile males had a CY spigot on each PMS, whereas adult males either had a CY spigot or, …

Utilizing Crispr/Cas9 Gene Editing Determine The Role Of Cnr1 During Zebrafish Development, Camme Sells

[Archive] Belmont University Research Symposium (BURS)

In a world where cannabinoids are being used more frequently as for medicinal purposes, the importance for research on cannabinoid biology is steadily increasing. The goal of this study was to gain a greater understanding of the effect cannabinoids have on developing zebrafish embryos. Work in this area of developmental biology could give way to an understanding of the safety of a human fetus during pregnancy with a mother who uses cannabis medicinal therapy. The CRISPR/Cas 9 system was utilized in this study to genetically edit the zebrafish embryo genome to remove the function of the Cannabinoid Receptor 1 ( …

Enhancement Of Nkx3.2 Expression After Twist1b And Twist2 Knockdown In Zebrafish, Peyton Yearick

[Archive] Belmont University Research Symposium (BURS)

Somitogenesis is a developmental event patterned by cellular signaling during gastrulation, ending around 30-hours post fertilization. Somites are segmented regions of mesoderm along the dorsal side of the body which form paraxial mesoderm in Danio rerio, zebrafish. Somite development results in the formation of the subcompartment called sclerotome, which differentiates to form the axial skeleton and associated tendons (Stickney et al., 2000). Further differentiation allows for sclerotome progenitor cell to develop into bone, muscle, tendon, and cartilage through sclerotome migration along the mesoderm. A portion of sclerotome cells migrate anterior towards the neural tube, where they will form the …

Expression Of Tgf-Beta Ligands During Sclerotome Development, Estela Williams

[Archive] Belmont University Research Symposium (BURS)

During early embryogenesis in vertebrates, somites are formed by the segmentation of the presomitic mesoderm (Gomez, Ozbudak, Wunderlich, Baumann. Lewis and Pourquie 2008). The somite possesses several domains within it and the ventral part of the somite is called the sclerotome. The sclerotome gives rise to the axial skeleton and tendons. Twist 1b and Twist 2 are genes that are expressed in the sclerotome during axial skeletal development. Our experiments were developed to help understand the roles of Twist1b and Twist2 in tendon development. By utilizing microinjection to alter gene expression, the Twist 1b and Twist 2 genes can be …

Using Crispr/Cas9 Gene Editing To Remove Twist2 To Determine Its Role During Somite Development In Zebrafish, Caleb Holdener

[Archive] Belmont University Research Symposium (BURS)

Embryonic Mesoderm is a precursor to various types of tissues, such as the somatic mesoderm. Somites are a segmented tissue that gives rise to the sclerotome in vertebrates. This sclerotome further develops into the axial skeleton (Stickney, Barresi, and Devoto 2000). Somites are easily observable under low power microscopy during fish development. In fish, great strides have been made in studying the developmental fate of somites (Stoiber, Haslett, and Sänger 1999; Stickney, Barresi, and Devoto 2000). Zebrafish have been studied in many labs due to their similarities with higher vertebrates. These advantages include their inexpensive cost of care, external fertilization, …

Twist1b Knockout Generation In Zebrafish Using Crispr-Cas9, Grace Hurley

[Archive] Belmont University Research Symposium (BURS)

The Twist gene is found in numerous organisms, including humans, mice and zebrafish (Yeo et al., 2009). It is a vital gene in development and is needed for the organism to survive (Yeo et al., 2009). Specifically, the Twist gene family aids in specialization and differentiation of mesodermal cells, telling somites, segmented blocks of mesoderm, what to become (Germanguz et al., 2007). Over time, the Twist gene has duplicated into paralogs, each having differences in their functions (Germanguz et al., 2007). Twist1b is a protein coding gene that is a paralog to the human Twist1 gene (ZFIN, 2020). In zebrafish, …

Scleraxis Expression Within The Axial Tendon Population In Zebrafish, Jessica Bernaba

[Archive] Belmont University Research Symposium (BURS)

Zebrafish are a well-established model organism for vertebrate development. During embryonic development, a pattern of segmentation of mesoderm yields somites, which give rise to specific cell fates. Somites, bilateral blocks of mesoderm along the neural tube in developing vertebrate embryos, subdivide into sclerotome as the vertebrate matures. Depending on migratory position, sclerotome cells give rise to various components of the axial skeletal system, including bone, cartilage, and tendon. Each fate is determined by gene expressions within the sclerotome and occupy distinct domains of the body plan. By method of in situ hybridization, we studied the expression of the scleraxis ( …

Utilizing Crispr/Cas9 Gene Editing Determine The Role Of Cnr1 During Zebrafish Development, Ellis Chase

[Archive] Belmont University Research Symposium (BURS)

The CRISPR-Cas 9 system is a process used to create mutations, insertions, or deletions within a desired gene. It can be used to create mutant organisms necessary to study the function and impact of a particular gene¹. Cannabinoids are active chemicals found in cannabis, the most common of which being cannabinol (CBD)². Cannabinoid receptors are an integral part of the endocannabinoid system, recognizing cannabinoid signals to promote physiological processes like pain sensation, memory, mood, and appetite³. Studying these cannabinoid receptors can provide insight into how cannabis can affect bodily processes.

Zebrafish are small translucent …

Exploring The Role Of Tgfbr2 During Zebrafish Sclerotome Development, Nooralhuda Feron

[Archive] Belmont University Research Symposium (BURS)

Zebrafish, Danio rerio, are used as a source model organism for human development of the tendons, bones, and ligaments. The sclerotome is an embryonic structure known as the somite. The sclerotome gives rise to axial bones and tendons. During development, different genes are involved and affect how these skeletal tissues develop. Specifically, within this project, the effects of Twist1b and Twist2 were studied to see their effects on tendon formation, as they are expressed within the developing sclerotome tissue. Transforming growth factor beta receptor type-2 (Tgfbr2) has been shown to be involved in tendon formation (Subramanian, Kanzaki, Galloway and …

Arid1a Haploinsufficiency Initiates Neural Crest Transformation In A Mouse Model Of Mycn-Driven Neuroblastoma, Kirby A. Wallace

Theses and Dissertations (ETD)

Mouse models of cancer are critical for developing therapeutic treatments for pediatric patients. Recent sequencing studies of neuroblastoma (NBL) patient tumors have uncovered precise deletions in the chromatin remodeler and tumor suppressor gene (TSG) ARID1A. Additional causal studies supported ARID1A’s candidacy as a putative 1p36 TSG in MYCN-driven NBL. This study aimed to causally test Arid1a loss during Mycn-driven NBL initiation through the development of a mouse model of high risk NBL.In this study, we modified a Mycn-driven mouse model of NBL to incorporate Cre mediated deletion of floxed Arid1a. Briefly, in freshly isolated embryonic day 9.5 primary trunk neural …

A Micro-Optic Stalk (Muos) System To Model The Collective Migration Of Retinal Neuroblasts, Stephanie Zhang, Miles Markey, Caroline D. Pena, Tadmiri Venkatesh, Maribel Vasquez

Publications and Research

Contemporary regenerative therapies have introduced stem-like cells to replace damaged neurons in the visual system by recapitulating critical processes of eye development. The collective migration of neural stem cells is fundamental to retinogenesis and has been exceptionally well-studied using the fruit fly model of Drosophila Melanogaster. However, the migratory behavior of its retinal neuroblasts (RNBs) has been surprisingly understudied, despite being critical to retinal development in this invertebrate model. The current project developed a new microfluidic system to examine the collective migration of RNBs extracted from the developing visual system of Drosophila as a model for the collective motile processes …

Inferring Tunicate Relationships And The Evolution Of The Tunicate Hox Cluster With The Genome Of Corella Inflata, M. B. Debiasse, William N. Colgan , '19, Lincoln J. Harris , '17, Bradley Justin Davidson , '91, J. F. Ryan

Biology Faculty Works

Tunicates, the closest living relatives of vertebrates, have served as a foundational model of early embryonic development for decades. Comparative studies of tunicate phylogeny and genome evolution provide a critical framework for analyzing chordate diversification and the emergence of vertebrates. Towards this goal, we sequenced the genome of Corella inflata (Ascidiacea, Phlebobranchia), so named for the capacity to brood self-fertilized embryos in a modified, “inflated” atrial chamber. Combining the new genome sequence for Co. inflata with publicly available tunicate data, we estimated a tunicate species phylogeny, reconstructed the ancestral Hox gene cluster at important nodes in the tunicate tree, and …

Variation In Brown Rat Cranial Shape Shows Directional Selection Over 120 Years In New York City, Emily E. Puckett, Emma Sherratt, Matthew Combs, Elizabeth J. Carlen, William Harcourt-Smith, Jason Munshi-South

Publications and Research

Urbanization exposes species to novel environments and selection pressures that may change morphological traits within a population. We investigated how the shape and size of crania and mandibles changed over time within a population of brown rats (Rattus norvegicus) living in Manhattan, New York, USA, a highly urbanized environment. We measured 3D landmarks on the cranium and mandible of 62 adult individuals sampled in the 1890s and 2010s. Static allometry explained approximately 22% of shape variation in crania and mandible datasets, while time accounted for approximately 14% of variation. We did not observe significant changes in skull size through time …