Biology Commons^™

A Combinatorial 5-Htr Expression Pattern Within The Ventral Projection Neurons Of The D. Melanogaster Olfactory Circuit., Mohd Freezely Ezzani Bin Mazri

Graduate Theses, Dissertations, and Problem Reports (ETD)

Neuromodulation allows neurons within a circuit to respond to stimuli from the environment according to the correct ecological value, context, and internal state of the animal. Serotonin (5-HT) is a neuromodulator that can generate different outcomes based on its target’s serotonin receptor (5-HTR) expression by affecting secondary messenger cascades. Within the Drosophila olfactory system, ventral projections neurons (vPN) express all five insect 5-HTR that project into two olfactory processing regions, the antennal lobe (AL) and the lateral horn (LH). The significance of this 5-HTR expression is unknown. In this study, I theorized the patterns of 5-HTR expression of vPNs. I …

Development And Characterization Of A Tissue-Specific Reporter Line For Monitoring Circadian Clock Transcriptional Activity, Lilyan Mather

Master's Theses

Daily rhythms in physiology and behavior are produced by a circadian system consisting of a central clock located in the brain and peripheral clocks located in various peripheral tissues. Circadian clocks track time of day through the presence of a molecular clock that functions as a transcriptional-translational feedback loop. In the nucleus, the transcription factors CLOCK (CLK) and CYCLE (CYC) activate transcription of clock genes period (per) and timeless (tim). PER and TIM proteins slowly accumulate in the cytoplasm before entering the nucleus, where they inhibit CLK and CYC, and therefore repress their own transcription. Degradation of nuclear PER and …

Empowering Melatonin Therapeutics With Drosophila Models, Cassandra Millet-Boureima, Caroline C. Ennis, Jurnee Jamison, Shana Mcsweeney, Anna Park, Chiara Gamberi

Biology

Melatonin functions as a central regulator of cell and organismal function as well as a neurohormone involved in several processes, e.g., the regulation of the circadian rhythm, sleep, aging, oxidative response, and more. As such, it holds immense pharmacological potential. Receptor-mediated melatonin function mainly occurs through MT1 and MT2, conserved amongst mammals. Other melatonin-binding proteins exist. Non-receptor-mediated activities involve regulating the mitochondrial function and antioxidant cascade, which are frequently affected by normal aging as well as disease. Several pathologies display diseased or dysfunctional mitochondria, suggesting melatonin may be used therapeutically. Drosophila models have extensively been employed to study disease pathogenesis …

The Effect Of Optogenetically Activating Glia On Neuronal Function, Cecilia Pankau, Shelby Mccubbin, Robin L. Cooper

Biology Faculty Publications

Glia, or glial cells, are considered a vital component of the nervous system, serving as an electrical insulator and a protective barrier from the interstitial (extracellular) media. Certain glial cells (i.e., astrocytes, microglia, and oligodendrocytes) within the CNS have been shown to directly affect neural functions, but these properties are challenging to study due to the difficulty involved with selectively-activating specific glia. To overcome this hurdle, we selectively expressed light-sensitive ion channels (i.e., channel rhodopsin, ChR2-XXL) in glia of larvae and adult Drosophila melanogaster. Upon activation of ChR2, both adults and larvae showed a rapid contracture of body wall …

A Docked Mutation Phenocopies Dumpy Oblique Alleles Via Altered Vesicle Trafficking, Suresh Kandasamy, Kiley Couto, Justin Thackeray

Biology

The Drosophila extracellular matrix protein Dumpy (Dpy) is one of the largest proteins encoded by any animal. One class of dpy mutations produces a characteristic shortening of the wing blade known as oblique (dpyo), due to altered tension in the developing wing. We describe here the characterization of docked (doc), a gene originally named because of an allele producing a truncated wing. We show that doc corresponds to the gene model CG5484, which encodes a homolog of the yeast protein Yif1 and plays a key role in ER to Golgi vesicle transport. Genetic analysis is consistent with a similar role …

Ipla2-Via Is Required For Healthy Aging Of Neurons, Muscle, And The Female Germline In Drosophila Melanogaster, Surya Jyoti Banerjee, Adina Schonbrun, Sogol Eizadshenass, Shimshon Benji, Yaakov Tzvi Cantor, Liam Eliach, Matthew Lubin, Zev Narrowe, Jeremy Purow, Benjamin Shulman, Leib Wiener, Josefa Steinhauer

Faculty Publications - Biological Sciences

Neurodegenerative disease (ND) is a growing health burden worldwide, but its causes and treatments remain elusive. Although most cases of ND are sporadic, rare familial cases have been attributed to single genes, which can be investigated in animal models. We have generated a new mutation in the calcium-independent phospholipase A2 (iPLA2) VIA gene CG6718, the Drosophila melanogaster ortholog of human PLA2G6/PARK14, mutations in which cause a suite of NDs collectively called PLA2G6-associated neurodegeneration (PLAN). Our mutants display age-related loss of climbing ability, a symptom of neurodegeneration in flies. Although phospholipase activity commonly is presumed to underlie iPLA2VIA function, locomotor decline …

Unbiased Automated Quantitation Of Ros Signals In Live Retinal Neurons Of Drosophila Using Fiji/Imagej, Prajakta Deshpande, Neha Gogia, Anuradha Venkatakrishnan Chimata, Amit Singh

Biology Faculty Publications

Numerous imaging modules are utilized to study changes that occur during cellular processes. Besides qualitative (immunohistochemical) or semiquantitative (Western blot) approaches, direct quantitation method(s) for detecting and analyzing signal intensities for disease(s) biomarkers are lacking. Thus, there is a need to develop method(s) to quantitate specific signals and eliminate noise during live tissue imaging. An increase in reactive oxygen species (ROS) such as superoxide (O2•-) radicals results in oxidative damage of biomolecules, which leads to oxidative stress. This can be detected by dihydroethidium staining in live tissue(s), which does not rely on fixation and helps prevent stress on tissues. However, …

Genetic Innovation Through Gene Duplication In Drosophila, Seyedeh Mirsalehi

Biology Dissertations - Archive

Among several mechanisms, whole gene duplication is one of the major sources for the generation of new genes. A large portion of young genes that emerged through whole gene duplication in Drosophila are specifically expressed in the testes and fast evolving, i.e., their proteins are changing very fast between species under positive selection. In this dissertation, we have studied the rates of duplications and functions of newly duplicated genes in three different projects to understand the evolutionary pressures/drivers of this gene innovation. Chapter 1 is an introductory section. In Chapter 2, by using a comprehensive, detailed phylogenomic study, we examined …

Late Embryogenesis Abundant Proteins And Mitochondrial Membranes From An Animal Extremophile: Insights Into Severe Water Stress, John Marcus Anderson

LSU Doctoral Dissertations

This dissertation focuses on two major topics germane to mechanisms by which animals tolerate extreme water stress. First, the impact of transgenic expression of late embryogenesis abundant proteins (LEA) on water stress tolerance of the fruit fly Drosophila melanogaster was investigated. The overall aim was to extend current understanding of the protective properties of LEA proteins documented with isolated cells to a desiccation-sensitive organism during exposure to drying and hyperosmotic stress. Four lines of D. melanogaster were created that expressed transgenes encoding selected LEA proteins originally identified in embryos of the extremophile Artemia franciscana. After 80% tissue water loss, …

A Forward Genetic Screen To Identify Human Genes Of Interest And Their Roles In Ovarian Cancer, Susan A. Ihejirika

Honors College Theses

Drosophila melanogaster, fruit flies, are very important for modeling and studying human diseases. This study identifies human genes of interest and their contributions to epithelial ovarian carcinogenesis and progression as well as the roles orthologs of these genes play in Drosophila melanogaster. This is important because ovarian cancer is the most common cause of death among the gynecological cancers. This identification of genes was carried out using a forward genetic screen employing the widely used GMR-Gal4 driver/UAS-transgene system. The GMR-Gal4 driver is commonly utilized to express transgenes in the developing eye of the fruit fly. Transgenes that are expressed using …

Rna Modification Landscape And Its Contribution To Egg Production, Ian Thomas Rapisarda

Legacy Theses & Dissertations (2009 - 2024)

Germline stem cells differentiate into mature egg or sperm cells that go through the process of fertilization which gives rise to all sexually reproducing organisms. During this process of differentiation, germ cells undergo a switch from mitosis to meiosis that allows for proper development and specification of the future gamete. The mechanisms that facilitate this shift from mitosis to meiosis, however, are not well understood. To gain insight into this process, we used Drosophila oogenesis as our model. To identify what RNA modifications are present during oogenesis we genetically enriched for each stage of development and performed mass spectrometry. We …

The Role Of The Dal Neurons In Modulating Circadian Rhythms In Olfactory Short-Term Memory In Drosophila Melanogaster, Cooper Ruwe

Honors Theses

Depressed short-term memory (STM) abilities during non-adaptive times of the day can significantly impact those who work occupations that require peak levels of cognitive functioning around the clock. While much work has gone into understanding the endogenous clock and circadian rhythms, there is still much to learn about the neural circuity that underlies the daily rhythms that define these regular oscillations in STM performance. The DAL neurons in the Drosophila brain are part of the circadian network and innervate the mushroom bodies (MBs), the species’ olfactory learning center, making them compelling candidates to be involved in circadian circuitry for olfactory …

Investigation Into The Akirin Subcellular Localization And Developmental Interactions, Riley Wildeman

Master of Science in Integrative Biology Theses

Akirin is a highly conserved nuclear protein ubiquitously expressed in nonfungal eukaryotic species and is known to be involved in many biological processes such as embryonic muscle patterning and innate immune function. Akirin functionality is essential for the prevention of disease and developmental defects. This investigation focused on both the subcellular localization of Akirin based on a putative nuclear localization signal and the genetic interaction between Akirin and the Nucleosome Remodeling Deacetylase (NuRD) complex. Through transfection of Drosophila S2 cells with Akirin mutant constructs we were able to visualize the localization patterns of Akirin and how the patterns relate to …

Modeling Neoplastic Growth In Renal Cell Carcinoma And Polycystic Kidney Disease, Cassandra Millet-Boureima, Stephanie He, Thi Bich Uyen Le, Chiara Gamberi

Biology

Renal cell carcinoma (RCC) and autosomal dominant polycystic kidney disease (ADPKD) share several characteristics, including neoplastic cell growth, kidney cysts, and limited therapeu-tics. As well, both exhibit impaired vasculature and compensatory VEGF activation of angiogenesis. The PI3K/AKT/mTOR and Ras/Raf/ERK pathways play important roles in regulating cystic and tumor cell proliferation and growth. Both RCC and ADPKD result in hypoxia, where HIF-α signaling is activated in response to oxygen deprivation. Primary cilia and altered cell metabolism may play a role in disease progression. Non-coding RNAs may regulate RCC carcinogenesis and ADPKD through their varied effects. Drosophila exhibits remarkable conservation of the …

Rna In Situ Hybridization For Detecting Gene Expression Patterns In The Abdomens And Wings Of Drosophila Species, Mujeeb Olushola Shittu, Tessa Steenwinkel, William Dion, Nathan Ostlund, Komal Raja, Thomas Werner

Michigan Tech Publications, Part 1

RNA in situ hybridization (ISH) is used to visualize spatio-temporal gene expression patterns with broad applications in biology and biomedicine. Here we provide a protocol for mRNA ISH in developing pupal wings and abdomens for model and non-model Drosophila species. We describe best practices in pupal staging, tissue preparation, probe design and synthesis, imaging of gene expression patterns, and image-editing techniques. This protocol has been successfully used to investigate the roles of genes underlying the evolution of novel color patterns in non-model Drosophila species.

Effect Of Temperature On Heart Rate For Phaenicia Sericata And Drosophila Melanogaster With Altered Expression Of The Trpa1 Receptors, Nicole T. Marguerite, Jate Bernard, Douglas A. Harrison, David Harris, Robin L. Cooper

Biology Faculty Publications

The transient receptor potential (TrpA—ankyrin) receptor has been linked to pathological conditions in cardiac function in mammals. To better understand the function of the TrpA1 in regulation of the heart, a Drosophila melanogaster model was used to express TrpA1 in heart and body wall muscles. Heartbeat of in intact larvae as well as hearts in situ, devoid of hormonal and neural input, indicate that strong over-expression of TrpA1 in larvae at 30 or 37 °C stopped the heart from beating, but in a diastolic state. Cardiac function recovered upon cooling after short exposure to high temperature. Parental control larvae (UAS-TrpA1) …

Experimental Use Of Drosophila To Model Proliferation Of Sars-Cov-2 Infected Cells, Valerie Wicks

Undergraduate Honors College Theses 2016-

Drosophila have been vectors for scientific research since the beginning of the 20th century. Their successful use within experiments that investigated mendelian genetics inspired their continuous application to biological research. This includes topics that detailed what is now known as innate immunity. The innate immune system of Drosophila is a first line of defense against pathogens. Innate immunity has been extensively researched and has reserved responses between Drosophila and mammals. Specifically, Drosophila have been experimentally targeted to draw conclusions on human infecting viruses such as Human Immunodeficiency Virus (HIV) and Severe Acute Respiratory Syndrome Coronavirus (SARS-Cov-1). Such experiments modeled viral …

Functional Genetic Approaches To Provide Evidence For The Role Of Toolkit Genes In The Evolution Of Complex Color Patterns In Drosophila Guttifera, Mujeeb Olushola Shittu

Dissertations, Master's Theses and Master's Reports

Toolkit genes are set of genes that orchestrate the development of basic body plan of animals, and they are highly conserved in all animals. The co-option of the toolkit genes into the pigmentation pathway has led to the evolution of novel species. This study focuses on understanding how the complex color patterns in animals develop by using the Drosophila species in the quinaria group as models. We developed an mRNA in situ hybridization (ISH) protocol, which allowed us to study gene expression patterns in the abdomen of developing pupae of non-model Drosophila species (Chapter 2). Through ISH, we found that …

Describing A Putative Corollary Discharge Circuit In Drosophila, Kaitlyn Nicole Boone

Graduate Theses, Dissertations, and Problem Reports (ETD)

Corollary discharge (CD) circuits provide critical information about movement and behavior to provide context to sensory processing. However, to date, there has not been a comprehensive study of CD circuits at a single-cell level. In this thesis, I aimed to resolve the connectivity of ascending histaminergic neurons, consisting of two pairs, the mesothoracic pair (MsAHNs) and metathoracic pair (MtAHNs) at a single-cell level and characterize contexts of activation. Using transgenic techniques, connectomics and transcriptomics, we identify neural populations receiving input from the AHNs and neural populations with significant output to the AHNs. We explored where the AHNs predominantly receive synaptic …

The Validation Of Predicted Biological Age Of Drosophila Melanogaster From Combined Statistical Modeling, Lauren Francis

Masters Theses

"Sleep is essential for maintaining a healthy body and mind and is associated with aging and aging related diseases. There are individual differences in fly as well as human sleep behavior and lifespan. Between and within individuals, sleep varies in characteristics including consolidation, rhythmicity, continuity, duration, and more. Various evidence in the literature suggests there are many molecular pathways involved with aging and they may be different for individuals. Our research is interested in a possible restorative mechanism of sleep and the ramifications of that mechanism to aging. We have developed two predictive models of aging using the fruit fly …

Drosophila Model To Study Muscle Atrophy, Aaron Aghai

Master of Science in Integrative Biology Theses

Muscle atrophy (MA) is a phenomenon of muscle mass loss due to accelerated protein degradation in muscle fibers. Some pathological conditions, such as chronic inflammation or cancer, induce accelerated MA, which complicates medical treatment, hampers recovery of fragile patients, and ultimately can be the cause of a patient’s death. To gain better control over MA, more information is required about the whole spectrum of genetic factors that can influence MA.

Drosophila provides an excellent platform for genetic screening, although it has somewhat limited utility for MA research since insect muscles lack the level of plasticity found in mammalian muscles. We …

Dissecting Dystrophin's Roles, Subcellular Organization, And Functional Network In Drosophila Oogenesis, Mina Amini Moghaddam

Theses and Dissertations

Mutations in the giant actin-membrane linker protein Dystrophin (Dys) are the cause of Duchenne Muscular Dystrophy (DMD). In the Drosophila model system, the highly conserved Dys protein is not required for viability but is required for proper wing and ovarian development. Dys mutations produce two visible phenotypes: posterior crossveins are detached from the longitudinal veins, and in oogenesis the developing eggs fail to elongate properly. This provides an opportunity to explore the less-understood cellular and developmental roles of Dys and gain a new insight into how tissues adopt their correct shapes.

Animals from worms to flies to humans express multiple …

Determining The Genetic Control Of Neural Tube Malformation Through Genetic Interactions With Idgf3, Elli N. Fox

Honors Projects

Genetic mutations disrupting human neural tube formation can lead to birth defects such as spina bifida and anencephaly. Defects can result in lack of neural tube closure in either the caudal (spina bifida) or cranial (anencephaly) regions. Little is known about the genes that cause these malformations. Researchers have been using the model organism Drosophila melanogaster in an attempt to determine genes responsible for neural tube malformations. Recently, an ortholog of human chitin-like protein, imaginal disc growth factor 3 (Idgf3), has been identified as important in the proper formation of Drosophila egg dorsal appendages. However, the molecular mechanism responsible for …

Direct Evidence Of Missing Mepsps Using Ca2-Sensor Imaging, Petar Gajic

Biological Sciences

Following an action potential in the presynaptic neuron there is evoked release of neurotransmitter into the synapse which activates ionotropic transmembrane receptors on the postsynaptic membrane that cause depolarizations in voltage that get recorded as excitatory postsynaptic potentials (EPSPs). In the absence of an action potential there is spontaneous release of neurotransmitter that postsynaptically gets recorded as miniature excitatory postsynaptic potentials (mEPSPs). According to the quantal hypothesis, postulated by Bernard Katz, the mEPSPs are allor- none changes in potential caused by a single quantum of neurotransmitter, which when added up create EPSPs. Following studies have found that these two modes …

The Development Of Complex Abdominal Spot Patterns In Three Drosophila Species, William A. Dion

Dissertations, Master's Theses and Master's Reports

Complex color patterns like the spots on leopards and butterfly wings beg the question – how did these traits evolve? To understand the evolution of novel morphologies, we need to study the differences in morphogenesis between closely related species. Here, I examine and compare the development of complex abdominal spot patterns among three species of Drosophila closely related in evolutionary time. Through in situ hybridization, I have characterized the expression patterns of three pigmentation genes among the fruit fly species. Additionally, I have built upon previous work in our lab regarding the regulation of pigmentation by beginning to develop an …

The Effect Of Ph On Synaptic Transmission At The Neuromuscular Junction In Drosophila Melanogaster, Catherine Elizabeth Stanley

Theses and Dissertations--Biology

Synaptic transmission is the main avenue of neuronal communication and can be affected by a multitude of factors, both intracellularly and extracellularly. The effects of pH changes on synaptic transmission have been studied for many years across many different models. Intracellular acidification at the presynaptic terminal is known to occur with increased neuronal activity and can also occur in pathological conditions. The effects of these pH alterations are therefore an important area of study. Here, intracellular acidification using either propionic acid or the ammonium chloride pre-pulse technique was examined for the effects on both spontaneous and evoked synaptic transmission at …

Actin Regulation And Furrow Dynamics During Early Drosophila Embryogenesis, Yi Xie

Electronic Theses and Dissertations

Drosophila embryogenesis starts with a single nucleus undergo 13 rounds of nuclear divisions called syncytial cycles. Staring at cycle 10 when nuclei migrate to the surface of the embryo, massive and dynamic cortical actin structures and cleavage furrow formations occur. How actin regulators coordinate into an organized network directing three-dimension actin structures in the developing organisms is an unsolved question. Here, I present an in-depth characterization of actin cap dynamics: the actin caps go through expansion, stabilization, elongation and fragmentation phases in each cycle. Arp2/3 is the major contributor to actin cap formation. The functions of 7 different actin and …

The 14-3-3 (Ywha) Proteins In Signalling And Development Of The Fruit Fly, Drosophila Melanogaster, Santanu De

Biology Faculty Articles

The 14-3-3 (YWHA or Tyrosine 3-Monooxygenase/Tryptophan 5-Monooxygenase Activation proteins) are a family of highly conserved, homologous proteins critical to diverse cellular events including cell cycle, signal transduction and embryonic development. Various species-specific isoforms of 14-3-3 exist, encoded by separate genes. They are expressed in a wide variety of organisms ranging from plants to animals, including the fruit fly or Drosophila melanogaster. Drosophila is one of the most universally accepted model systems to study complex cellular mechanisms of signalling and development. However, regulation of these processes in fruit flies by the 14-3-3 proteins have not been entirely understood. This mini …

Analysis Of The Temporal Patterning Of Notch Downstream Targets During Drosophila Melanogaster Egg Chamber Development, Molly A. Rowe

Honors College Theses

Living organisms require complex signaling interactions and proper regulation of these interactions to influence biological processes. Of these complex networks, one of the most distinguished is the Notch pathway. Dysregulation of this pathway often results in defects during organismal development and can be a causative mechanism for initiation and progression of cancer. Despite previous research entailing the importance of this signaling pathway and the organismal processes that it is involved in, less is known concerning the major Notch downstream targets, especially the onset and sequence in which they are modulated during normal development. As timing of regulation may be linked …

Invertebrate Retinal Progenitors As Regenerative Models In A Microfluidic System, Caroline D. Pena, Stephanie Zhang, Robert Majeska, Tadmiri Venkatesh, Maribel Vazquez

Publications and Research

Regenerative retinal therapies have introduced progenitor cells to replace dysfunctional or injured neurons and regain visual function. While contemporary cell replacement therapies have delivered retinal progenitor cells (RPCs) within customized biomaterials to promote viability and enable transplantation, outcomes have been severely limited by the misdirected and/or insuffcient migration of transplanted cells. RPCs must achieve appropriate spatial and functional positioning in host retina, collectively, to restore vision, whereas movement of clustered cells differs substantially from the single cell migration studied in classical chemotaxis models. Defining how RPCs interact with each other, neighboring cell types and surrounding extracellular matrixes are critical to …