Cell and Developmental Biology Commons^™

A Study Of Tissue-Specific Rho Signaling Activation During Tubular Organ Formation In Drosophila, Charlotte Jean-Louise Adams

LSU Master's Theses

Several organs within the human body are tubular in structure. In the event of a malfunction in the development of these tubular structures, maladies like spina bifida, anencephaly, and several other defects can arise. To understand the mechanisms of tubular organ formation, we use the Drosophila embryonic salivary gland (SG) as a model. During embryogenesis, SG cells undergo a series of cell shape changes, like apical constriction, to accommodate the developing gland. Apical constriction is regulated in part by the signal of the secreted ligand Folded gastrulation (Fog) and its G protein-coupled receptor (GPCR) at the cell membrane, which leads …

Brown Anole (Anolis Sagrei) Hoxa5: Insights Into The Divergence Of Hoxa5 Gene Expression And Regulation Across Evolutionarily Divergent Gnathostome Vertebrates, Jennifer A. Lange, Amber L. Rittgers, Adam Davis

Georgia Journal of Science

Hox genes are evolutionarily conserved developmental regulatory genes that function, in part, to pattern the anterior-posterior (AP) axis of organs and organ systems during animal embryonic development. Hoxa5, specifically, is shown to be expressed in the spinal cord, somites, or transient compartments giving rise to the vertebrae and ribs, developing gut, lungs, and limbs of the mouse (Mus musculus). The cis-regulatory elements (CREs), or short DNA sequences, that direct Hoxa5 expression in these embryonic domains have been mapped and functionally tested in the mouse as well. Similar Hoxa5 expression patterns have been observed in chicken ( …

Trimeric Complex Interactions Of Antp-Tbp With Tfiieb And Exd Are Involved In The Genetic Control Of Drosophila Melanogaster, Gustavo Jiménez Mejía, Ruben De Jesus Montalvo Mendez, Claudia Dalila Altamirano Torres, Diana Reséndez Pérez

Research Symposium

Background: Homeoproteins are transcriptional factors (TFs) that shape animal body axes during development. These TFs are highly conserved and represent one of the most fascinating groups of regulatory molecules. Reports shown the multiplicity of interactions in hox proteins, as complexes trimeric involved to transcriptional activity. The study of trimeric complexes in Hox interactome will allow the better understanding of Hox genetic regulation during embryonic development.

Methodology: Using a new combination BiFCFRET approach performed in HEK293, the quantification was performed by FRETTY of ImageJ. Fly crosses were incubated at 25ºC on standard yeast-agar-cornmeal medium. Embryo cuticle preparations were carried out according …

Tata-Box Binding Protein Interacts With Antp, Scr, Ubx And Abdb Through Their N-Terminal Domains, Rubén Montalvo Méndez, Gustavo Jiménez Mejía, Diana Reséndez Pérez

Research Symposium

Background: Hox proteins are transcriptional factors (TFs) that define segment identity during embryonic development regulating specific target genes. These TFs interact with cofactors for DNA specificity and other TFs to regulate gene expression, which include basal transcriptional machinery members like BIP2, Med19, TFIIEβ, M1BP and TBP. Since TBP glutamine homopeptide (PolyQ) act as an interaction domain involved transcriptional regulation, we analyzed if TBP interact with Antp, Scr, Ubx and AbdB through its PolyQ region.

Methods: We used Bimolecular Fluorescent Complementation (BiFC) to determine TBP interaction with Antp, Scr, Ubx and AbdB as well as the implication of their homeodomain (HD) …

Trimeric Interaction Of Antp-Ubx With Tbp And Homeoprotein Exd In The Genetic Control Of Development In D. Melanogaster, Alely Cecilia Villarreal-Puente, Claudia Dalila Altamirano-Torres, Diana Resendez-Perez

Research Symposium

Homeoproteins specify body segments along the anteroposterior axis during embryo development; they bind DNA through the homeodomain (HD). Homeoproteins bind to similar and repetitive target sequences on DNA, raising the question of how they achieve functional specificity. Homeoproteins form complexes with proteins through the HDs or other regions. Antennapedia (Antp) and Ultrabithorax (Ubx) have an important role in conferring thorax identity; also they are involved in a repression transcriptional mechanism: Ubx binds to Antp promoter repressing its expression. We analyzed if Ubx and Antp perform protein-protein interaction as a regulation mechanism and if the complex Antp-Ubx are involved in trimeric …

Examining Pi3k-Signaling-Dependent Regulation Of Lens Organelle Free Zone Formation Via Immunolocalization And Immunoblotting In Chick Embryos, Rifah Gheyas, A. Sue Menko

Computational Medicine Center Faculty Papers

The elimination of lens organelles during development, required for mature lens function, is an autophagy-dependent mechanism induced through suppression of PI3K signaling. Here, we present a protocol for investigating the signaling pathways responsible for induction of the formation of this lens organelle free zone. We describe steps for preparation of lens organ culture and use of signaling pathway inhibitors. We then detail procedures for analyzing their impact using both confocal microscopy imaging of immunolabeled lens cryosections and immunoblot approaches. For complete details on the use and execution of this protocol, please refer to Gheyas et al. (2022).

Canine Testes Thin Sections Culture​, Nguyen Khoi Nguyen, Ramanpreet Singh

2023 Symposium

Canine testes thin section culture

Testes tissue culture systems would provide a tool to elucidate spermatogenesis mechanisms, with the aim of genetic preservation of mammals, especially endangered species. Our experiment aims to develop a culture system capable of producing viable mammalian sperm cells in vitro.

Dogs were chosen as the model organism because testes are readily available. Canine testes were obtained from a local veterinary clinic. Thin sections were generated using a commercial electric slicer. They then were cleaned using Dulbecco’s Phosphate-Buffered Saline (DPBS) supplemented with antibiotics then cultured in a modified Tissue Culture Medium 199 (TCM-199). Sections were …

Canine Testes Thin Sections Culture, Nguyen Khoi Nguyen, Ramanpreet Singh

2023 Symposium

Canine testes thin section culture

Testes tissue culture systems would provide a tool to elucidate spermatogenesis mechanisms, with the aim of genetic preservation of mammals, especially endangered species. Our experiment aims to develop a culture system capable of producing viable mammalian sperm cells in vitro.

Dogs were chosen as the model organism because testes are readily available. Canine testes were obtained from a local veterinary clinic. Thin sections were generated using a commercial electric slicer. They then were cleaned using Dulbecco’s Phosphate-Buffered Saline (DPBS) supplemented with antibiotics then cultured in a modified Tissue Culture Medium 199 (TCM-199). Sections were …

Appendage Abnormalities In Spiders Induced By An Alternating Temperature Protocol In The Context Of Recent Advances In Molecular Spider Embryology, Teresa Napiorkowska, Julita Templin, Pawel Napiorkowski, Mark A. Townley

Faculty Publications

In the literature there are numerous reports of developmental deformities in arthropods collected in their natural habitat. Since such teratogenically affected individuals are found purely by chance, the causes of their defects are unknown. Numerous potential physical, mechanical, chemical, and biological teratogens have been considered and tested in the laboratory. Thermal shocks, frequently used in teratological research on the spider Eratigena atrica, have led to deformities on both the prosoma and the opisthosoma. In the 2020/2021 breeding season, by applying alternating temperatures (14 °C and 32 °C, changed every 12 h) for the first 10 days of embryonic development, …

Mirna Regulation Of Skeletal Muscle Hypertrophy, Maslyn Greene

All Dissertations

Muscle tissue is vital to animal survival and economically essential to livestock producers. Reduced muscle growth can occur from various reasons, including genetics, nutrition, and the pre and postnatal environment. Understanding the mechanisms behind muscle growth can aid in developing strategies to increase muscle mass. Several methods to increase muscle mass in livestock animals are currently being pursued, including nutrition, hormonal therapy, and genetic selection. One newly developing avenue is using miRNA technologies to change the expression of protein-coding genes. miRNA are a type of RNA that regulate over half of protein-coding gene expression. Here we evaluate muscle characteristics such …

Mob Family Proteins And Tricornered Kinase Are Required To Form Dorsal Appendages Of The Drosophila Eggshell, Keala H. D. Watson

UNLV Theses, Dissertations, Professional Papers, and Capstones

To understand how complex tissues are formed, we need to understand how cells communicate with other cells to build structures. In Drosophila melanogaster, epithelial cells form tubular structures to shape the dorsal appendages on the eggshells. One signaling pathway needed for this event is the Hippo-like kinase intracellular pathway. I focus on two key components in this pathway: the NDR kinase Tricornered (Trc) and Mob proteins. In eukaryotes, Mob proteins are known to interact with NDR kinases to promote their activation. The Hippo- like pathway involves the NDR kinase Trc; however, the specific Mob protein(s) that interact with Trc …

Striped Expression Of Leucine-Rich Repeat Proteins Coordinates Cell Intercalation And Compartment Boundary Formation In The Early Drosophila Embryo, Chloe A. Kuebler, Adam C. Pare

Biological Sciences Faculty Publications and Presentations

Planar polarity is a commonly observed phenomenon in which proteins display a consistent asymmetry in their subcellular localization or activity across the plane of a tissue. During animal development, planar polarity is a fundamental mechanism for coordinating the behaviors of groups of cells to achieve anisotropic tissue remodeling, growth, and organization. Therefore, a primary focus of developmental biology research has been to understand the molecular mechanisms underlying planar polarity in a variety of systems to identify conserved principles of tissue organization. In the early Drosophila embryo, the germband neuroectoderm epithelium rapidly doubles in length along the anterior-posterior axis through a …

Functional Analysis Of The Gamma-Aminobutyric Acid Type A Receptor Subunit Alpha-1 (Gabra1) Gene During Zebrafish Development., Nayeli Gabriela Reyes-Nava

Open Access Theses & Dissertations

The GABRA1 gene encodes for the alpha-1 (α1) subunit of the Gamma-Aminobutyric acid type A receptor (GABAAR), which are the primary modulators of synaptic inhibition in the central nervous system (CNS). Alpha-1 subunits are essential for maintaining the normal function of native receptors and contribute to over 60% of all GABAARs in the CNS. Remarkably, a broad spectrum of neurodevelopmental and epilepsy-associated disorders have been linked with mutations in the GABRA1 gene. However, the developmental, behavioral, and molecular mechanisms underlying GABRA1-associated epileptic disorders remain to be fully understood. Hence, the overarching goal of this dissertation is to investigate the behavioral …

A Novel Role For Enos In Regulating Lymphatic Valve Development During Embryogenesis, Drishya Iyer

USF Tampa Graduate Theses and Dissertations

Lymphedema is a disease that occurs when lymph flow is impaired, resulting in tissue swelling, fibrosis, chronic inflammation and recurrent secondary infections. Lymphatic valves play a critical role in maintaining unidirectional lymph flow and evidence for valve defects have been reported in lymphedema patients. The lack of drugs that can correct lymphatic valve defects warrants a better understanding of the molecular regulators of lymphatic valve development and maintenance. Lymphatic valves first develop during embryogenesis in response to mechanotransduction signaling pathways triggered by oscillatory lymph flow. Since eNOS (gene name: Nos3) is a well characterized mechanotransduction signaling molecule in blood vessels, …

Anillin And Dunk: The Conserved And Non-Conserved Players Regulating Myosin Dynamics In Drosophila Cleavage, Jiayang Chen

Dartmouth College Ph.D Dissertations

Cleavage is a common process during early embryogenesis. Similar to conventional cytokinesis, Drosophila cellularization, a unique form of cleavage, requires precise regulation of non-muscle myosin II ("myosin") at cleavage furrows, where myosin forms an interconnected basal array before reorganizing into individual cytokinetic rings. However, how myosin is regulated through both conserved and cellularization-specific mechanisms during this special form of cytokinesis remains unclear.

In my first project, I investigated the molecular mechanisms by which the cellularization-specific zygotic gene dunk regulates myosin at the basal array. I identified anillin (Scraps in Drosophila), a conserved scaffolding protein in cytokinesis, as the primary …

Evaluation Of Nutrient Digestibility Of Weaned Calves From Early And Late Shedding Dams, Jennifer Keele

Theses and Dissertations

Researchers have investigated several factors that could alter fetal growth, including nutrient restriction (Valiente et al., 2021), hair shedding (Gray et al., 2011), and extreme hot and cold temperatures (Davidson et al., 2022). Hot temperatures and increased humidity percentages in the southeast United States caused researchers to investigate the hair coats of Angus cattle in the commercial production setting. An improvement in fiber digestibility and calf birth and weaning weights has been observed in Angus dams that shed 50% of the winter hair coat by May (Gray et al., 2011; Burnett et al., 2021). Our objective of this experiment was …

Understanding The Regulatory Context And Mechanism Of Hlh-25 Mediated Transcriptional Repression, Dagmawi Mamo

Masters Theses, 2020-current

The HES family of transcription factors are primarily repressors of transcription that play critical roles in mammalian embryogenesis by regulating cell differentiation and proliferation. Proper functioning of HES proteins is crucial in humans, as mutations or misexpression of HES1 is associated with lung, pancreas, colon, and other cancers. HES1 may mediate short-range or long-range repression through DNA-binding and or protein-protein interactions with other factors. However, prior genetic studies have not fully elucidated how the context-specific binding of HES1, or its C. elegans ortholog HLH-25, promotes the repression of transcription. Characterization of the cis-regulatory context and mechanism of HLH-25 mediated repression …

Microglia Refine Early Multisensory Circuits In The Lateral Cortex Of The Inferior Colliculus In A Complement-Dependent Manner, Shaida Hamidi

Masters Theses, 2020-current

Microglia cells (MGCs), the resident macrophage of the brain, play a pivotal role in synaptic pruning of newly established circuits during early critical periods. Classical complement cascade signaling enables MGCs to identify unnecessary or underutilized contacts and specifically target them for removal and subsequent degradation. Such recognition and engulfment behaviors require receptor-ligand interactions, whereby “eat me” tags (complement component 3, C3) bind their cognate receptor (complement receptor 3, CR3) expressed on MGCs. Compromised complement signaling in certain systems results in under-pruning or unrefined map configurations, and is thought perhaps to underlie certain autism-related behavioral phenotypes (e.g. sensory hypersensitivities, decreased social …

Growth Plate Cartilage: Understanding The Contribution Of Adhesion To Column Formation And Matrix Structure, Sydney E. Greer

Theses & Dissertations

Throughout fetal and adolescent development, bone growth is regulated by fine-tuned and controlled maturation of chondrocytes through a cartilaginous template called the growth plate. Bone growth rate is controlled through cell enlargement and extracellular matrix deposition, while the polarized arrangement of proliferative chondrocytes into columns aligned with the long axis of the bone potentiate growth. Chondrocytes are surrounded by a complex three-dimensional arrangement of matrix molecules, all of which are secreted by chondrocytes and assembled/remodeled to support the biological functions of the cell. Adhesion receptors found on the cell membrane of chondrocytes are crucial to the organization of matrix proteins …

Measuring Energy Demands Of Dynamic Remodeling Events During Embryogenesis In Live Drosophila Embryos, Grayson Young

Biological Sciences Undergraduate Honors Theses

Embryonic development is a complex process requiring dynamic cell movements to create complex tissue structures. We assume that these embryonic dynamic remodeling events are highly energy intensive and that this energy expenditure is mediated by mitochondria. However, this has been difficult to demonstrate in live embryos. Convergent extension of the Drosophila neuroectoderm is a promising system to study the bioenergetics of tissue remodeling because it changes from a static to a dynamically remodeling tissue with temporal precision. We hypothesize that the switch from static to dynamic development at the onset of convergent extension will be accompanied by an increase in …

Mitochondrial Roles In Developmentally Programmed Heart Disease, Eli John Louwagie

Dissertations and Theses

Offspring of diabetic and obese mothers (ODOM) have greater risks of heart disease at birth and later in life. However, prevention is hindered because underlying mechanisms are poorly understood. Mounting studies in the Developmental Origins of Health and Disease field suggest that mitochondria play key roles in developmentally programmed heart disease similar to the roles they play in cardiomyopathy in adults with diabetes and obesity. However, whether mitochondria are responsible for the short[1]and long-term cardiac disease seen in ODOM remains unknown. Here, we sought to delineate the roles of mitochondria in the hearts of ODOM, determine whether mitochondria are playing …

Transcriptional Regulation Of Stem Cell Function In The Drosophila Central Nervous System, Amalia S. Parra

Biology ETDs

Development of multicellular organisms depends on coordination of various molecular mechanisms to ensure proper tissue size and structure. This is accomplished through intrinsic and extrinsic cues that regulate cell growth and proliferation both spatially and temporally. These well-coordinated signals are present from early developmental stages and remain active throughout development to ensure tissue structure is appropriate for function. This is particularly important in early cell types that form specialized lineages such as stem cells or those found throughout the body such as epithelia. While much progress has been made in understanding growth regulatory mechanisms, several fundamental aspects remain unknown. This …

Examining Transcriptional Regulators During Muscle Development In Drosophila Melanogaster, Chaamy Yapa

Student Theses and Dissertations

In Drosophila melanogaster embryos, a distinct approach to study the transcriptional regulation is to examine the larval somatic muscle development. Transcription factors are essential regulatory proteins that help to control gene expression and respond to signaling pathways and various cues. Today, there are at least twenty transcription factors that have been discovered to contribute to the development of the 30 distinct larval somatic muscles in each abdominal hemisegment of Drosophila melanogaster. Several studies have already been conducted on muscle regulatory transcription factors including midline and apterous. These transcription factors were shown to control the development of muscles through mutant …

The Effects Of Glycolytic Mutations In Drosophila Melanogaster Muscle Development, Coco Lim

Student Theses and Dissertations

Muscle atrophy, or muscle wasting, is caused due to lack of physical activity for an extended period of time, due to muscle diseases (such as muscle dystrophies), cancer chemotherapies, and aging. It is also extensively found on astronauts after spaceflight, particularly missions of long durations. Muscle cells are dependent on different metabolic pathways to optimize Adenosine triphosphate (ATP) production to compensate for muscle exertion. Glycolysis converts glucose into ATP producing pyruvate, which can be sent into the citric acid cycle or converted to lactate (lactic acid). Muscles preferentially use lactate production, despite the fact that fewer molecules of ATP are …

Functional Analysis Of Daxx In Tumorigenesis Of Pancreatic Neuroendocrine Tumors And Embryonic Development, Chang Sun

Dissertations and Theses (Open Access)

Death domain-associated protein 6 (Daxx) is a histone chaperone specific to Histone 3.3 (H3.3). DAXX interacts with ATRX forming a chromatin remodeling complex, which deposits H3.3 into telomeric and pericentric region of the genome. The importance of Daxx was manifested in embryonic development. The loss of Daxx leads to early lethality in mouse embryos around E6.5. Moreover, sequencing studies have revealed the importance of DAXX in human tumors. Mutually exclusive mutations in DAXX and ATRX occur in about 30% of pancreatic neuroendocrine tumors (PanNETs). Although lots of progress has been made in studying functions of DAXX, we still do not …

Adipocytes And Innate Immunity In Systemic Sclerosis, Nancy Wareing

Dissertations and Theses (Open Access)

Systemic sclerosis (SSc; scleroderma) is a chronic systemic autoimmune and connective tissue disorder characterized by vasculopathy, autoimmune phenomena, and widespread fibrosis. Skin thickening and tightening is the cardinal feature of SSc and is responsible, in part, for the considerable morbidity of this disease. There are currently no targeted treatments for skin manifestations in SSc, primarily due to our fragmented understanding of its pathophysiologic mechanisms. In PART I, we report a previously unappreciated link between aberrant expression of the developmental gene sine oculis homeobox homolog 1 (SIX1) in skin-associated adipocytes in SSc skin and the early loss of dermal white adipose …

Genetic Regulation Of Müllerian Duct Regression, Malcolm Moses

Dissertations and Theses (Open Access)

Mammals, including humans, develop progenitor tissues for both male and female reproductive tract organs before they fully differentiate into a male or female tract. The progenitor tissue for the male reproductive tract is known as the Wolffian duct (WD), and the progenitor tissue for the female reproductive tract is the Müllerian duct (MD). The WD further differentiates into the vas deferens, epididymis, and seminal vesicle, while the MD differentiates into the oviduct, uterus and upper vagina. An essential step in sex differentiation for males is the regression of the MD. This regression initiates with anti-Müllerian hormone (Amh) transcription …

Investigation The Plugging Behavior Of Virus Filters, Iman Abdulqader Saab

Graduate Theses and Dissertations

A virus filtration step is integral to the manufacturing of biopharmaceuticals to ensure viral safety. A virus filter is a single-use device that uses a size-based separation process and has a unique structure with minimal defects, so that contaminating virus particles cannot pass through the membrane pores, while therapeutic molecules can. The development of novel antibodies (Abs), including significant increases in product titers, is frequently challenged by virus filter fouling, making a better understanding of the underlying mechanisms essential. This thesis focused investigating of the effect of prefilter types, buffer type and salt content on virus filtration performance. The impact …

Gonads Without Glp-1: Silencing Glp-1 In The Male Somatic Gonad In Caenorhabditis Elegans, Matthew Titus

Undergraduate Theses

In C. elegans, the gene glp-1 encodes for a Notch receptor called GLP-1, one of two found in C. elegans’ genome. The gene has been previously implicated in the development of the hermaphroditic germline as well as playing a role in the mitosis/meiosis decision. Genetic screening has further identified it as potentially playing a role in the development of the male somatic gonad, making it an ideal candidate for a reverse genetic. We did this by silencing glp-1 and observing if any alterations to the gonad’s phenotype occur.

Normally this could be done by performing a gene knockout. …

Understanding The Expression And Role Of Pros-1 In The Male Gonad Of C. Elegans, Jack Bozik

Undergraduate Theses

The gene pros-1 is a transcription factor that is highly expressed within neuronal sheath cells, glial cells, and excretory canal cells. pros-1 plays a role in cell determination of those cell types in the nematode C. elegans, which promotes organismal development. But the degree to which pros-1 presence is important is still not fully understood, because there are many genes involved in development that when mutated or damaged can result in unexpected phenotypes or even total loss of function to a certain developmental mechanism. What makes pros-1 valuable to research is that it is a functional homologue to a …