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- Title
- Effects of Serotonin Modulation on Methionine Sulfoxide Reductase Deficient Drosophila melanogaster.
- Creator
- Hamadeh, Ali, Binninger, David, Florida Atlantic University, Department of Biological Sciences, Charles E. Schmidt College of Science
- Abstract/Description
-
Methionine sulfoxide reductase (MSR) is an important antioxidant to help mitigate oxidative stress that contributes to age-associated neurodegenerative diseases, such as Alzheimer’s Disease and Parkinson’s Disease. In MSR deficient Drosophila melanogaster (fruit flies), larvae show a developmental delay like that seen when wild-type larvae are reared on nutrient deficit culture medium. These investigators further showed that serotonin levels were depressed in these nutrient deficient larvae....
Show moreMethionine sulfoxide reductase (MSR) is an important antioxidant to help mitigate oxidative stress that contributes to age-associated neurodegenerative diseases, such as Alzheimer’s Disease and Parkinson’s Disease. In MSR deficient Drosophila melanogaster (fruit flies), larvae show a developmental delay like that seen when wild-type larvae are reared on nutrient deficit culture medium. These investigators further showed that serotonin levels were depressed in these nutrient deficient larvae. The overarching aim of this study was to better understand the role of serotonin in MSR regulated physiology. Supplementing food with serotonin partially rescued the slower mouth hook movements (MHM) observed in the MSR-deficient flies. However, supplementation with serotonin altering drugs that cross the blood brain barrier (5-hydroxytryptophan, fluoxetine, or paravi chlorophenylalanine) did not rescue MHM and caused impairments to the growth of larvae during development. This study indicates that serotonin regulates feeding behavior partially through the regulation of MSR production but acts independently to regulate development.
Show less - Date Issued
- 2021
- PURL
- http://purl.flvc.org/fau/fd/FA00013761
- Subject Headings
- Drosophila melanogaster, Methionine sulfoxide reductase, Serotonin
- Format
- Document (PDF)
- Title
- GENETIC SCREENS IDENTIFY NOVEL REGULATORS OF SLEEP AND METABOLISM IN DROSOPHILA MELANOGASTER.
- Creator
- Murakami, Kazuma N., Keene, Alex C., Florida Atlantic University, Department of Biological Sciences, Charles E. Schmidt College of Science
- Abstract/Description
-
Proper regulation of sleep and metabolism are critical to the survival of all organisms. In humans, dysregulation of sleep is linked to metabolic syndrome, including hypertension, hyperglycemia and hyperlipidemia. However, the mechanisms regulating interactions between sleep and metabolism are poorly understood. Although the fruit fly, Drosophila melanogaster, bears little anatomical resemblance to humans, it shares similar genetics essential in understanding normal development and disease in...
Show moreProper regulation of sleep and metabolism are critical to the survival of all organisms. In humans, dysregulation of sleep is linked to metabolic syndrome, including hypertension, hyperglycemia and hyperlipidemia. However, the mechanisms regulating interactions between sleep and metabolism are poorly understood. Although the fruit fly, Drosophila melanogaster, bears little anatomical resemblance to humans, it shares similar genetics essential in understanding normal development and disease in humans. From humans to flies, many disease-related genes and pathways are highly conserved, rendering the fruit fly ideal to understanding the interactions between sleep and metabolism. Therefore, using the fruit fly provides a framework for understanding how genes function between sleep and metabolism. During starvation, both humans and rats reduce their sleep. Similarly, previous studies have shown that fruit flies also suppress sleep to forage for food, further showing that sleep and metabolism are intricately tied to one another and that they are highly conserved across species. To further explore the interactions between sleep and metabolism, I have conducted multiple genetic screens to identify novel regulators of sleep-metabolism interactions. These experiments led to the identification of the mRNA binding protein translin (trsn) as being required for starvation-induced sleep suppression. A second screen that targeted metabolic genes from a genome-wide association study identified the ion channel accessory protein uncoordinated 79 (unc79) as a critical regulator of both sleep duration and starvation resistance. The genes function in different regions of the brain and suggest complex neural circuitry is likely to underlie regulation of sleep metabolism interactions. Taken together, a mechanistic understanding of how different genes function to regulate sleep in flies will further our understanding of how sleep and metabolism is regulated in humans.
Show less - Date Issued
- 2021
- PURL
- http://purl.flvc.org/fau/fd/FA00013722
- Subject Headings
- Drosophila melanogaster, Sleep, Genetic screening
- Format
- Document (PDF)
- Title
- HISTAMINERGIC AND NOCICEPTIVE GROOMING IN DROSOPHILA MELANOGASTER: AN ANALYSIS OF THE MOLECULAR MECHANISMS AND A BEHAVIORAL RESPONSE TO NOXIOUS CHEMICAL STIMULI.
- Creator
- John, Ciny, Dawson-Scully, Ken, Murphey, Rodney, Florida Atlantic University, Charles E. Schmidt College of Science, Department of Biological Sciences
- Abstract/Description
-
Insect grooming has various functions, including defense against parasites and pathogens, cleaning of dust particles, and maintenance of sensory receptors. The hierarchy of grooming behavior suggests that cleaning one body part is more crucial than the other, the priority order more specifically being eyes, antennae, abdomen, then wings, followed by the thorax. Histamine is an extensively studied neurotransmitter found in the central nervous system of many animals. In Drosophila, histamine is...
Show moreInsect grooming has various functions, including defense against parasites and pathogens, cleaning of dust particles, and maintenance of sensory receptors. The hierarchy of grooming behavior suggests that cleaning one body part is more crucial than the other, the priority order more specifically being eyes, antennae, abdomen, then wings, followed by the thorax. Histamine is an extensively studied neurotransmitter found in the central nervous system of many animals. In Drosophila, histamine is found in both the peripheral and central nervous systems and is necessary for visual and mechanosensory behaviors. Histamine-gated chloride channel 1 (HisCl1) and Ora transientless (Ort) are two characterized histamine receptors, both of which are vital for visual signaling in the fly.
Show less - Date Issued
- 2019
- PURL
- http://purl.flvc.org/fau/fd/FA00013321
- Subject Headings
- Drosophila melanogaster, Grooming behavior in animals, Nociception, Histaminergic mechanisms
- Format
- Document (PDF)
- Title
- Functional Stress Resistance: The Role of Protein Kinase G in Modulating Neuronal Excitability in Caenorhabditis Elegans and Drosophila Melanogaster.
- Creator
- Kelly, Stephanie Suzanne, Dawson-Scully, Ken, Florida Atlantic University, Charles E. Schmidt College of Science, Department of Biological Sciences
- Abstract/Description
-
Diseases such as epilepsy, pain, and neurodegenerative disorders are associated with changes in neuronal dysfunction due to an imbalance of excitation and inhibition. This work details a novel electroconvulsive seizure assay for C. elegans using the well characterized cholinergic and GABAergic excitation and inhibition of the body wall muscles and the resulting locomotion patterns to better understand neuronal excitability. The time to recover normal locomotion from an electroconvulsive...
Show moreDiseases such as epilepsy, pain, and neurodegenerative disorders are associated with changes in neuronal dysfunction due to an imbalance of excitation and inhibition. This work details a novel electroconvulsive seizure assay for C. elegans using the well characterized cholinergic and GABAergic excitation and inhibition of the body wall muscles and the resulting locomotion patterns to better understand neuronal excitability. The time to recover normal locomotion from an electroconvulsive seizure could be modulated by increasing and decreasing inhibition. GABAergic deficits and a chemical proconvulsant resulted in an increased recovery time while anti-epileptic drugs decreased seizure duration. Successful modulation of excitation and inhibition in the new assay led to the investigation of a cGMP-dependent protein kinase (PKG) which modulates potassium (K+) channels, affecting neuronal excitability, and determined that increasing PKG activity decreases the time to recovery from an electroconvulsive seizure. The new assay was used as a forward genetic screening tool using C. elegans and several potential genes that affect seizure susceptibility were found to take longer to recover from a seizure. A naturally occurring polymorphism for PKG in D. melanogaster confirmed that both genetic and pharmacological manipulation of PKG influences seizure duration. PKG has been implicated in stress tolerance, which can be affected by changes in neuronal excitability associated with aging, so stress tolerance and locomotor behavior in senescent flies was investigated. For the first time, PKG has been implicated in aging phenotypes with high levels of PKG resulting in reduced locomotion and lifespan in senescent flies. The results suggest a potential new role for PKG in seizure susceptibility and aging.
Show less - Date Issued
- 2019
- PURL
- http://purl.flvc.org/fau/fd/FA00013225
- Subject Headings
- Caenorhabditis elegans, Drosophila melanogaster, Cyclic GMP-Dependent Protein Kinases, Seizures
- Format
- Document (PDF)
- Title
- Genetic and Neural Mechanisms Regulating the Interaction Between Sleep and Metabolism in Drosophila Melanogaster.
- Creator
- Yurgel, Maria E., Keene, Alex C., Florida Atlantic University, Charles E. Schmidt College of Science, Department of Biological Sciences
- Abstract/Description
-
Dysregulation of sleep and metabolism has enormous health consequences. Sleep loss is linked to increased appetite and insulin insensitivity, and epidemiological studies link chronic sleep deprivation to obesity-related disorders. Interactions between sleep and metabolism involve the integration of signalling from brain regions regulating sleep, feeding, and metabolism, as well as communication between the brain and peripheral organs. In this series of studies, using the fruit fly as a model...
Show moreDysregulation of sleep and metabolism has enormous health consequences. Sleep loss is linked to increased appetite and insulin insensitivity, and epidemiological studies link chronic sleep deprivation to obesity-related disorders. Interactions between sleep and metabolism involve the integration of signalling from brain regions regulating sleep, feeding, and metabolism, as well as communication between the brain and peripheral organs. In this series of studies, using the fruit fly as a model organism, we investigated how feeding information is processed to regulate sleep, and how peripheral tissues regulate sleep through the modulation of energy stores. In order to address these questions, we performed a large RNAi screen to identify novel genetic regulators of sleep and metabolism. We found that, the mRNA/DNA binding protein, Translin (trsn), is necessary for the acute modulation of sleep in accordance with feeding state. Flies mutant for trsn or selective knockdown of trsn in Leucokinin (Lk) neurons abolishes starvation-induced sleep suppression. In addition, genetic silencing of Lk neurons or a mutation in the Lk locus also disrupts the integration between sleep and metabolism, suggesting that Lk neurons are active during starvation. We confirmed this hypothesis by measuring baseline activity during fed and starved states. We found that LHLK neurons, which have axonal projections to sleep and metabolic centers of the brain, are more active during starvation. These findings suggest that LHLK neurons are modulated in accordance with feeding state to regulate sleep. Finally, to address how peripheral tissues regulate sleep, we performed an RNAi screen, selectively knocking down genes in the fat body. We found that knockdown of Phosphoribosylformylglycinamidine synthase (Ade2), a highly conserved gene involved the biosynthesis of purines, regulates sleep and energy stores. Flies heterozygous for two Ade2 mutations are short sleepers and this effect is partially rescued by restoring Ade2 to the fly fat body. These findings suggest Ade2 functions within the fat body to promote both sleep and energy storage, providing a functional link between these processes. Together, the experimental evidence presented here provides an initial model for how the peripheral tissues communicate to the brain to modulate sleep in accordance with metabolic state.
Show less - Date Issued
- 2018
- PURL
- http://purl.flvc.org/fau/fd/FA00013163
- Subject Headings
- Drosophila melanogaster, Sleep, Metabolism
- Format
- Document (PDF)
- Title
- Alternative Biological Roles of Methionine Sulfoxide Reductases in Drosophila melanogaster.
- Creator
- Wilson, Kelsey, Binninger, David, Florida Atlantic University, Charles E. Schmidt College of Science, Department of Biological Sciences
- Abstract/Description
-
The oxidation of methionine (Met) into methionine sulfoxide (met-(o)) leads to deleterious modifications to a variety of cellular constituents. These deleterious alterations can be reversed by enzymes known as methionine sulfoxide reductases (Msr). The Msr (MsrA and MsrB) family of enzymes have been studied extensively for their biological roles in reducing oxidized Met residues back into functional Met. A wide range of studies have focused on Msr both in vivo and in vitro using a variety of...
Show moreThe oxidation of methionine (Met) into methionine sulfoxide (met-(o)) leads to deleterious modifications to a variety of cellular constituents. These deleterious alterations can be reversed by enzymes known as methionine sulfoxide reductases (Msr). The Msr (MsrA and MsrB) family of enzymes have been studied extensively for their biological roles in reducing oxidized Met residues back into functional Met. A wide range of studies have focused on Msr both in vivo and in vitro using a variety of model organisms. More specifically, studies have noted numerous processes affected by the overexpression, under expression, and silencing of MsrA and MsrB. Collectively, the results of these studies have shown that Msr is involved in lifespan and the management of oxidative stress. More recent evidence is emerging that supports existing biological functions of Msr and theorizes the involvement of Msr in numerous biological pathways.
Show less - Date Issued
- 2018
- PURL
- http://purl.flvc.org/fau/fd/FA00005980
- Subject Headings
- Drosophila melanogaster, Methionine Sulfoxide Reductases, Oxidative stress
- Format
- Document (PDF)
- Title
- Neuroprotection during acute hyperthermic stress: Role of the PKG pathway in neurons and glia in the protection of neural function in Drosophila melanogaster.
- Creator
- Krill, Jennifer L., Dawson-Scully, Ken, Florida Atlantic University, Charles E. Schmidt College of Science, Department of Biological Sciences
- Abstract/Description
-
The human brain functions within a narrow range of temperatures and variations outside of this range incur cellular damage and death and, ultimately, death of the organism. Other organisms, like the poikilotherm Drosophila melanogaster, have adapted mechanisms to maintain brain function over wide ranges in temperature and, if exposed to high temperatures where brain function is no longer supported, these animals enter a protective coma to promote survival of the organism once the acute...
Show moreThe human brain functions within a narrow range of temperatures and variations outside of this range incur cellular damage and death and, ultimately, death of the organism. Other organisms, like the poikilotherm Drosophila melanogaster, have adapted mechanisms to maintain brain function over wide ranges in temperature and, if exposed to high temperatures where brain function is no longer supported, these animals enter a protective coma to promote survival of the organism once the acute temperature stress is alleviated. This research characterized the role of different neuronal cell types, including glia, in the protection of brain function during acute hyperthermia, specifically looking at two protective pathways: the heat shock protein (HSP) pathway and the cGMP-dependent protein kinase G (PKG) pathway. Whole animal behavioral assays were used in combination with tissue-specific genetic manipulation of protective pathways to determine the specific cell types sufficient to confer protection of neuronal function during acute hyperthermia. Using the neuromuscular junction (NMJ) preparation, calcium imaging techniques were combined with pharmacological and genetic manipulations to test the hypothesis that alterations in ion channel conductance via endogenous mechanisms regulating the cellular response to high temperature stress alter neuronal function. Expression of foraging RNAi to inhibit PKG expression in neurons or glia demonstrated protection of function during acute hyperthermia measured behaviorally through the extension of locomotor function. This extension of function with the tissue-specific inhibition of PKG was also confirmed at the cellular level using the genetically encoded calcium indicator (GECI), GCaMP3, to image calcium dynamics at the NMJ, where preparations expressing foraging RNAi could continue to elicit changes in calcium dynamics in response to stimulation. Over the course of this study, the mechanism underlying a novel glial calcium wave in the peripheral nervous system was characterized in order to elucidate glia’s role in the protection of neuronal function during acute hyperthermia.
Show less - Date Issued
- 2018
- PURL
- http://purl.flvc.org/fau/fd/FA00013026
- Subject Headings
- Cyclic GMP-Dependent Protein Kinases, Neuroprotection, Hyperthermia, Heat shock proteins, Drosophila melanogaster
- Format
- Document (PDF)
- Title
- Characterizing electroconvulsive seizure recovery time in the invertebrate model systems Caenorhabditis elegans and Drosophila melanogaster.
- Creator
- Risley, Monica G., Dawson-Scully, Ken, Florida Atlantic University, Charles E. Schmidt College of Science, Department of Biological Sciences
- Abstract/Description
-
Seizures are a symptom of epilepsy, characterized by spontaneous firing due to an imbalance of excitatory and inhibitory features. While mammalian seizure models receive the most attention, the simplicity and tractability of invertebrate model systems, specifically C. elegans and D. melanogaster, have many advantages in understanding the molecular and cellular mechanisms of seizure behavior. This research explores C. elegans and D. melanogaster as electroconvulsive seizure models to...
Show moreSeizures are a symptom of epilepsy, characterized by spontaneous firing due to an imbalance of excitatory and inhibitory features. While mammalian seizure models receive the most attention, the simplicity and tractability of invertebrate model systems, specifically C. elegans and D. melanogaster, have many advantages in understanding the molecular and cellular mechanisms of seizure behavior. This research explores C. elegans and D. melanogaster as electroconvulsive seizure models to investigate methods to both modulate and better understand seizure susceptibility. A common underlying feature of seizures in mammals, worms, and flies involves regulating excitation and inhibition. The C. elegans locomotor circuit is regulated via well characterized GABAergic and cholingeric motoneurons that innervate two rows of dorsal and ventral body wall muscles. In this research, we developed an electroconvulsive seizure assay which utilizes the locomotor circuit as a behavioral read out of neuronal function. When inhibition is decreased in the circuit, for example by decreasing GABAergic input, we find a general increase in the time to recovery from a seizure. After establishing the contribution of excitation and inhibition to seizure recovery, we explored a ubiquitin ligase, associated with comorbidity of an X-linked Intellectual Disorder and epilepsy in humans, and established that the worm homolog, eel-1, contributes to seizure susceptibility similarly to the human gene. Next, we investigated a cGMP-dependent protein kinase (PKG) that functions in the nervous system of both worms and flies and determined that increasing PKG activity, decreases the time to recovery from an electroconvulsive seizure. These experiments suggest a potential novel role for a major protein, PKG, in seizure susceptibility and that the C. elegans and D. melanogaster electroconvulsive seizure assays can be used to investigate possible genes involved in seizure susceptibility and future therapeutic to treat epilepsy.
Show less - Date Issued
- 2018
- PURL
- http://purl.flvc.org/fau/fd/FA00005954
- Subject Headings
- Seizures, Epilepsy, Drosophila melanogaster, Caenorhabditis elegans
- Format
- Document (PDF)
- Title
- Functional roles of L1-Cam/Neuroglian in the nervous system of Drosophila Melanogaster.
- Creator
- Kudumala, Sirisha, Godenschwege, Tanja A., Florida Atlantic University, Charles E. Schmidt College of Science, Department of Biological Sciences
- Abstract/Description
-
Neuronal cell adhesion molecules of L1 family play a critical role in proper nervous system development. Various mutations on human L1-CAM that lead to severe neurodevelopmental disorders like retardation, spasticity etc. termed under L1 syndrome. The vertebrr their roles in axon pathfinding, neurite extension and cell migration, howeverate L1CAM and its homolog in Drosophila, neuroglian (nrg) have been well studied fo, much less is known about the mechanisms by which they fine tune synaptic...
Show moreNeuronal cell adhesion molecules of L1 family play a critical role in proper nervous system development. Various mutations on human L1-CAM that lead to severe neurodevelopmental disorders like retardation, spasticity etc. termed under L1 syndrome. The vertebrr their roles in axon pathfinding, neurite extension and cell migration, howeverate L1CAM and its homolog in Drosophila, neuroglian (nrg) have been well studied fo, much less is known about the mechanisms by which they fine tune synaptic connectivity to control the development and maintenance of synaptic connections within neuronal circuits. Here we characterized the essential role of nrg in regulating synaptic structure and function in vivo in a well characterized Drosophila central synapse model neuron, the Giant Fiber (GF) system. Previous studies from our lab revealed that the phosphorylation status of the tyrosine in the Ankyrin binding FIGQY motif in the intracellular domain of Nrg iscrucial for synapse formation of the GF to Tergo-Trochanteral Motor neuron (TTMn) synapse in the GF circuit. The present work provided us with novel insights into the role of Nrg-Ank interaction in regulating Nrg function during synapse formation and maintenance. By utilizing a sophisticated Pacman based genomic rescue strategy we have shown that dynamic regulation of the Neuroglian–Ankyrin interaction is required to coordinate transsynaptic development in the GF–TTMn synapse. In contrast, the strength of Ankyrin binding directly controls the balance between synapse formation and maintenance at the NMJ. Human L1 pathological mutations affect different biological processes distinctively and thus their proper characterization in vivo is essential to understand L1CAM function. By utilizing nrg14;P[nrg180ΔFIGQY] mutants that have exclusive synaptic defects and the previously characterized nrg849 allele that affected both GF guidance and synaptic function, we were able to analyze pathological L1CAM missense mutations with respect to their effects on guidance and synapse formation in vivo. We found that the human pathological H210Q, R184Q and Y1070C, but not the E309K and L120V L1CAM mutations affect outside-in signaling via the FIGQY Ankyrin binding domain which is required for synapse formation and not for axon guidance while L1CAM homophilic binding and signaling via the ERM motif is essential for axon guidance in Drosophila.
Show less - Date Issued
- 2014
- PURL
- http://purl.flvc.org/fau/fd/FA00004131, http://purl.flvc.org/fau/fd/FA00004131
- Subject Headings
- Cell adhesion molecules, Cellular signal transduction, Cognitive neuroscience, Cognitive neuroscience, Drosophila melanogaster, Molecular neurobiology
- Format
- Document (PDF)
- Title
- Development of a novel assay for in vivo screening of neuromodulatory drugs and targeted disruption of cholinergic synaptic transmission in Drosophila melanogaster.
- Creator
- Mejia, Monica, Charles E. Schmidt College of Science, Department of Biological Sciences
- Abstract/Description
-
Finding novel compounds that affect neuronal or muscular function is of great interest, as they can serve as potential pharmacological agents for a variety of neurological disorders. For instance, conopeptides have been developed into powerful drugs like the painkiller PrialtTM. Most conopeptides, however, have yet to be characterized, revealing the need for a rapid and straightforward screening method. We have designed a novel bioassay, which allows for unbiased screening of biological...
Show moreFinding novel compounds that affect neuronal or muscular function is of great interest, as they can serve as potential pharmacological agents for a variety of neurological disorders. For instance, conopeptides have been developed into powerful drugs like the painkiller PrialtTM. Most conopeptides, however, have yet to be characterized, revealing the need for a rapid and straightforward screening method. We have designed a novel bioassay, which allows for unbiased screening of biological activity of compounds in vivo against numerous molecular targets on a wide variety of neurons and muscles in a rapid and straightforward manner. For this, we paired nanoinjection of compounds with electrophysiological recordings from the Giant Fiber System of Drosophila melanogaster, which mediates the escape response of the fly.
Show less - Date Issued
- 2013
- PURL
- http://purl.flvc.org/fcla/dt/3362560
- Subject Headings
- Drosophila melanogaster, Genetics, Drosophila melanogaster, Life cycles, Insects, Physiology, Developmental neurobiology, Neural transmission, Cholinergic mechanisms
- Format
- Document (PDF)
- Title
- Phenotypic and behavioral effects of methionine sulfoxide reductase deficiency and oxidative stress in Drosophila melanogaster.
- Creator
- Mulholland, Kori., Charles E. Schmidt College of Science, Department of Biological Sciences
- Abstract/Description
-
Harman's theory of aging proposes that a buildup of damaging reactive oxygen species (ROS) is one of the primary causes of the deleterious symptoms attributed to aging. Cellular defenses in the form of antioxidants have evolved to combat ROS and reverse damage; one such group is the methionine sulfoxide reductases (Msr), which function to reduce oxidized methionine. MsrA reduces the S enantiomer of methionine sulfoxide, Met-S-(o), while MsrB reduces the R enantiomer, Met-R-(o). The focus of...
Show moreHarman's theory of aging proposes that a buildup of damaging reactive oxygen species (ROS) is one of the primary causes of the deleterious symptoms attributed to aging. Cellular defenses in the form of antioxidants have evolved to combat ROS and reverse damage; one such group is the methionine sulfoxide reductases (Msr), which function to reduce oxidized methionine. MsrA reduces the S enantiomer of methionine sulfoxide, Met-S-(o), while MsrB reduces the R enantiomer, Met-R-(o). The focus of this study was to investigate how the absence of one or both forms of Msr affects locomotion in Drosophila using both traditional genetic mutants and more recently developed RNA interference (RNAi) strains. Results indicate that lack of MsrA does not affect locomotion. However, lack of MsrB drastically reduces rates of locomotion in all age classes. Furthermore, creation of an RNAi line capable of knocking down both MsrA and MsrB in progeny was completed.
Show less - Date Issued
- 2013
- PURL
- http://purl.flvc.org/fcla/dt/3362558
- Subject Headings
- Drosophila melanogaster, Genetics, Aging, Molecular aspects, Oxidative stress, Mitochondrial pathology, Cellular signal transduction, Oxidation-reduction reaction, Biochemical markers, Mutation (Biology)
- Format
- Document (PDF)
- Title
- Neuroprotection during anoxic-stress in Drosophila melanogaster: the role of PKG pathway on protection of function and survival.
- Creator
- Benasayag Meszaros, Raquel, Charles E. Schmidt College of Science, Department of Biological Sciences
- Abstract/Description
-
Anoxia is characterized by an absence of oxygen supply to a tissue (Dawson- Scully et al., 2010). Unlike humans, Drosophila melanogaster is an organism that can survive low oxygen levels for hours without showing any pathology (Lutz et al., 2003) Under anoxia, the fruit fly loses locomotive activity, resulting in an anoxic coma (Haddad et al., 1997). In this study we investigate the influence of five variables for anoxic tolerance in adult Drosophila: 1) anoxic environment (gas vs. drowning),...
Show moreAnoxia is characterized by an absence of oxygen supply to a tissue (Dawson- Scully et al., 2010). Unlike humans, Drosophila melanogaster is an organism that can survive low oxygen levels for hours without showing any pathology (Lutz et al., 2003) Under anoxia, the fruit fly loses locomotive activity, resulting in an anoxic coma (Haddad et al., 1997). In this study we investigate the influence of five variables for anoxic tolerance in adult Drosophila: 1) anoxic environment (gas vs. drowning), 2) anoxia duration, 3) temperature (cold [3ÀC] or room temperature [21ÀC]), 4) age (young 2-9 days and old 35-39 days), and 5) PKG variation. Tolerance to anoxia is measured by the time of recovery and survival of the fruit fly from the anoxic coma. The results from this study show that short stress, low temperature, young age, and low PKG activity increased anoxic tolerance. Our findings will lay the foundation to investigate different variables, genes or pharmacological compounds that can modulate neuronal anoxic tolerance.
Show less - Date Issued
- 2013
- PURL
- http://purl.flvc.org/FAU/3360743
- Subject Headings
- Protein kinases, Oxidativie stress, Prevention, Oxidativie stress, Ecophysiology, Drosophila melanogaster, Life cycles
- Format
- Document (PDF)
- Title
- Comprehensive study of the ZAD family of zinc finger transcription factors in Drosophila melanogaster.
- Creator
- Krystel, Joseph., Charles E. Schmidt College of Science, Department of Biological Sciences
- Abstract/Description
-
The zinc finger associated domain (ZAD) family of transcription factors from Drosophila melanogaster is not well described in the literature, in part because it is very difficult to study by traditional mutagenesis screens. Bioinformatic studies indicate this is due to overlapping functions remaining after a recent evolutionary divergence. I set out to use in vitro-binding techniques to identify the characteristics of the ZAD family and test this theory. I have constructed glutathione S...
Show moreThe zinc finger associated domain (ZAD) family of transcription factors from Drosophila melanogaster is not well described in the literature, in part because it is very difficult to study by traditional mutagenesis screens. Bioinformatic studies indicate this is due to overlapping functions remaining after a recent evolutionary divergence. I set out to use in vitro-binding techniques to identify the characteristics of the ZAD family and test this theory. I have constructed glutathione S-transferase (GST)-ZAD domain chimeric proteins for use in pull down protein binding assays,and GST-Zinc finger (ZnF) array domain chimera for electrophoretic mobility shift assays (EMSA). Protein binding assays indicated two putative conserved interactors, similar to the analogous KRAB system in mammals. ... Competitive bindings were carried out to show a specificity of binding conferred by the identified conserved positions. While the consensus binding sites show relatively few similarities, the predicted target genes identified by the consensus binding sites show significant overlap. The nature of this overlap conforms to the known characteristics of the ZAD family but points to a more positive selection to maintain conservation of function.
Show less - Date Issued
- 2012
- PURL
- http://purl.flvc.org/FAU/3355627
- Subject Headings
- Cellular signal transduction, Drosophila melanogaster, Cytogenetics, Transcription factors, Zinc-finger proteins, Synthesis, Genetic transcription, Regulation, Gene expression
- Format
- Document (PDF)
- Title
- Highwire's characterization and signaling roles in Drosophila central synapse formation.
- Creator
- Rowland, Kimberly Diane., Charles E. Schmidt College of Science, Department of Biological Sciences
- Abstract/Description
-
The assembly and maintenance of central synapses is a complex process, requiring myriad genes and their products. Highwire is a large gene containing a RING domain, characteristic of ubiquitin E3 ligases. Highwire has been shown to restrain axon growth and control synaptogenesis at a peripheral synapse. Here I examine the roles of Highwire at a central synapse in the adult Drosophila Giant Fiber System (GFS). Highwire is indeed necessary for proper axonal growth as well as synaptic...
Show moreThe assembly and maintenance of central synapses is a complex process, requiring myriad genes and their products. Highwire is a large gene containing a RING domain, characteristic of ubiquitin E3 ligases. Highwire has been shown to restrain axon growth and control synaptogenesis at a peripheral synapse. Here I examine the roles of Highwire at a central synapse in the adult Drosophila Giant Fiber System (GFS). Highwire is indeed necessary for proper axonal growth as well as synaptic transmission in the GFS. Differences arise between the central synapse and the neuromuscular junction (NMJ), where highwire was initially characterized : expresion from the postsynaptic cell can rescue highwire synaptic defects, which is not seen at the NMJ. In addition, a MAP kinase signaling pathway regulated by highwire at the NMJ has differing roles at a central synapse. Wallenda MAPK can rescue not only the highwire anatomical phenotype but also the defects seen in transmission. Another distinction is seen here : loss of function basket and Dfos enhance the highwire anatomical phenotype while expression of dominant negative basket and Dfos suppress the highwire phenotype. As a result we have compared the signaling pathway in flies and worms and found that the NMJ in the two organisms use a parallel pathway while the central synapse uses a distinct pathway.
Show less - Date Issued
- 2012
- PURL
- http://purl.flvc.org/FAU/3352826
- Subject Headings
- Cellular control mechanisms, Cellular signal transduction, Cell differentiation, Gene expression, Genetic transcription, Transcription factors, Drosophila melanogaster, Cytogenetics
- Format
- Document (PDF)
- Title
- Identification and characterization of mutations in the Drosophila mitochondrial translation elongation factor iconoclast.
- Creator
- Trivigno, Catherine F., Charles E. Schmidt College of Science, Department of Biological Sciences
- Abstract/Description
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Mitochondrial disorders resulting from defects in oxidative phosphorylation are the most common form of inherited metabolic disease. Mutations in the human mitochondrial translation elongation factor GFM1 have recently been shown to cause the lethal pediatric disorder Combined Oxidative Phosphorylation Deficiency Syndrome (COXPD1). Children harboring mutations in GFM1 exhibit severe developmental, metabolic and neurological abnormalities. This work describes the identification and extensive...
Show moreMitochondrial disorders resulting from defects in oxidative phosphorylation are the most common form of inherited metabolic disease. Mutations in the human mitochondrial translation elongation factor GFM1 have recently been shown to cause the lethal pediatric disorder Combined Oxidative Phosphorylation Deficiency Syndrome (COXPD1). Children harboring mutations in GFM1 exhibit severe developmental, metabolic and neurological abnormalities. This work describes the identification and extensive characterization of the first known mutations in iconoclast (ico), the Drosophila orthologue of GFM1. Expression of human GFM1 can rescue ico null mutants, demonstrating functional conservation between the human and fly proteins. While point mutations in ico result in developmental defects and death during embryogenesis, animals null for ico survive until the second or third instar larval stage. These results indicate that in addition to loss-of-function consequences, point mutations in ico appear to produce toxic proteins with antimorphic or neomorphic effects. Consistent with this hypothesis, transgenic expression of a mutant ICO protein is lethal when expressed during development and inhibits growth when expressed in wing discs. In addition, animals with a single copy of an ico point mutation are more sensitive to acute hyperthermic or hypoxic stress. Removal of the positively-charged tail of the protein abolishes the toxic effects of mutant ICO, demonstrating that this domain is necessary for the harmful gain-of-function phenotypes observed in ico point mutants., Further, expression of GFP-tagged constructs indicates that the C-terminal tail enhances ectopic nuclear localization of mutant ICO, suggesting that mislocalization of the protein may play a role in the antimorphic effects of mutant ICO. Taken together, these results illustrate that in addition to loss-of-function effects, gain-of-function effects can contribute significantly to the pathology caused by mutation in mitochondrial translation elongation factors.
Show less - Date Issued
- 2010
- PURL
- http://purl.flvc.org/FAU/2705081
- Subject Headings
- Drosophila melanogaster, Cytogenetics, Mutation (Biology), Mitochondrial DNA, Cell metabolism, Cellular signal transduction, Oxidation, Physiological, Genetic transcription, Regulation
- Format
- Document (PDF)
- Title
- Identification of longitudinals lacking (LOLA) target genes in Drosophila melanogaster.
- Creator
- Qureshi, Bazila., Charles E. Schmidt College of Science, Department of Biological Sciences
- Abstract/Description
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Longitudinals lacking gene (LOLA) is a transcription factor that is involved in a variety of axon guidance decisions in Drosophila melanogaster nervous system. Besides having a role as an epigenetic silencer and in the programmed cell death in Drosophila's ovary, this gene is also an example of complex transcription unit. LOLA is a transcription repressor and can generate 17 DNA - binding isoforms, through alternative splicing, each containing distinct zinc-finger proteins. This unique...
Show moreLongitudinals lacking gene (LOLA) is a transcription factor that is involved in a variety of axon guidance decisions in Drosophila melanogaster nervous system. Besides having a role as an epigenetic silencer and in the programmed cell death in Drosophila's ovary, this gene is also an example of complex transcription unit. LOLA is a transcription repressor and can generate 17 DNA - binding isoforms, through alternative splicing, each containing distinct zinc-finger proteins. This unique DNAbinding binding sequence to which LOLA-ZFP binds has been determined for four of the lola isoforms F, J, P and K. Also, bioinformatics' tool approach has been taken to identify the target genes that are regulated by these four LOLA splice variants. Future work will be done for the five other LOLA isoforms to categorize their putative DNA-binding sequences and subsequently their protein interactions.
Show less - Date Issued
- 2010
- PURL
- http://purl.flvc.org/FAU/2684893
- Subject Headings
- Transcription factors, Cellular signal transduction, Zinc-finger proteins, Synthesis, Genetic transcription, Regulation, Drosophila melanogaster, Cytogenetics, Gene expression
- Format
- Document (PDF)
- Title
- Elucidation of the features of the zinc finger associated domain (ZAD) family of transportation factors in Drosophila melanogaster.
- Creator
- Krystel, Joseph., Charles E. Schmidt College of Science, Department of Biological Sciences
- Abstract/Description
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The zinc finger associated domain (ZAD) containing family of transcription factors is not well described in the literature, in part because it is very difficult to study by mutagenesis. We used in vitro-binding techniques to identify characteristics of the ZAD family, by constructing glutathione Stransferase (GST)-ZAD domain chimeric proteins for use in protein binding assays, and GST-Zinc finger array domain chimera for binding site selections. Protein binding assays indicated a possible...
Show moreThe zinc finger associated domain (ZAD) containing family of transcription factors is not well described in the literature, in part because it is very difficult to study by mutagenesis. We used in vitro-binding techniques to identify characteristics of the ZAD family, by constructing glutathione Stransferase (GST)-ZAD domain chimeric proteins for use in protein binding assays, and GST-Zinc finger array domain chimera for binding site selections. Protein binding assays indicated a possible shared cofactor, as seen in the analogous KRAB system in mammals. DNA binding assays have provided a consensus binding sequence for five of the ZAD proteins, consistent with previously reported work on ZAD and unpublished work on mammalian transcription factors. Research is ongoing with an additional ~50 ZAD proteins to more fully map the binding characters of ZAD proteins.
Show less - Date Issued
- 2009
- PURL
- http://purl.flvc.org/FAU/186768
- Subject Headings
- Cellular signal transduction, Drosophila melanogaster, Cytogenetics, Transcription factors, Zinc-finger proteins, Synthesis, Genetic transcription, Regulation, Gene expression
- Format
- Document (PDF)