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Title: A rapid colorimetric microassay to detect agonists/antagonists of protein kinase C based on adherence of EL-4.IL-2 cells.
Creator: Longley, Ross E., Harmody, Dedra K., Harbor Branch Oceanographic Institute
Date Issued: 1991
PURL: http://purl.flvc.org/FCLA/DT/3351944
Subject Headings: Protein kinase C, Colorimetric analysis, Diglycerides
Format: Document (PDF)

Title: Rapid neuroprotection from acute oxidative stress.
Creator: Caplan, Stacee Lee, Dawson-Scully, Ken, Milton, Sarah L., Graduate College
Date Issued: 2011-04-08
PURL: http://purl.flvc.org/fcla/dt/3164512
Subject Headings: Stress --physiology, Protein kinases, Neuroprotective agents
Format: Document (PDF)

Title: Staurosporine aglycone (K252-c) and arcyriaflavin A from the marine ascidian, Eudistoma sp.
Creator: Horton, P. A., Longley, Ross E., Harbor Branch Oceanographic Institute
Date Issued: 1994
PURL: http://purl.flvc.org/fau/fd/FA00007230
Subject Headings: Ascidians, Sea squirts, Protein kinase C, Cytotoxins, Indoles, Carbazoles
Format: Document (PDF)

Title: cGMP/PKG-regulated mechanisms of protection from low oxygen and oxidative stress.
Creator: Mahneva, Olena, Milton, Sarah L., Dawson-Scully, Ken, Florida Atlantic University, Charles E. Schmidt College of Science, Department of Biological Sciences
Abstract/Description: Stroke is one of the leading causes of human death in the United States. The debilitating effects of an ischemic stroke are due to the fact that mammalian neurons are highly susceptible to hypoxia and subsequent oxygen reperfusion. From studies in Drosophila melanogaster, cGMP-dependent Protein Kinase (PKG) enzyme is thought to affect anoxia tolerance by modifying the electrical current through potassium ion channels. In this research, two animal models were employed: Drosophila melanogaster...
Show moreStroke is one of the leading causes of human death in the United States. The debilitating effects of an ischemic stroke are due to the fact that mammalian neurons are highly susceptible to hypoxia and subsequent oxygen reperfusion. From studies in Drosophila melanogaster, cGMP-dependent Protein Kinase (PKG) enzyme is thought to affect anoxia tolerance by modifying the electrical current through potassium ion channels. In this research, two animal models were employed: Drosophila melanogaster and mammalian neurons exposed to stroke-like conditions. First, in vivo studies using Drosophila were performed to further our knowledge about the differences between the naturally occurring variants of the Drosophila foraging gene, which shows different protein levels of PKG. Mitochondrial density and metabolic activity between two fly genotypes exposed to anoxia and reoxygenation were compared. It was found that flies with less enzyme potentially showed mitochondrial biogenesis and higher metabolic rates upon reoxygenation. Next, in vivo studies where PKG enzyme was activated pharmacologically were performed; it was found that the activation of the cGMP/PKG pathway led to neuroprotection upon anoxia and reoxygenation. Furthermore, this model was translated into the in vitro model using Drosophila cells. Instead of anoxia and reoxygenation, hypoxia mimetics and hydrogen peroxide were used to induce cellular injury. After showing the cGMP/PKG pathway activation-induced cell protection, the potential downstream targets of the molecular signaling as well as underlying biochemical changes were assessed. It was found that mitochondrial potassium ion channels were involved in the protective signaling and the signaling modulated metabolic function. Furthermore, it was found that acidosis protected Drosophila cells from cell death, metabolic disruption, and oxidative stress. Finally, this research was translated to a mammalian in vitro model of neuronal damage upon stroke-like conditions; there, it was demonstrated that the cGMP/PKG pathway activation in rat primary cortical neurons and human cortical neurons was protective from low oxygen and acute oxidative stress. The results of this study lead to a better understanding of molecular mechanisms taking place during low oxygen and oxidative stresses. Consequently, this knowledge may be used to identify potential therapeutic targets and treatments that may prevent detrimental neurological effects of an ischemic stroke in humans.
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Date Issued: 2018
PURL: http://purl.flvc.org/fau/fd/FA00013013
Subject Headings: Stroke, Cyclic GMP-Dependent Protein Kinases, 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.
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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: Neuroprotection During Acute Oxidative Stress: Role of the PKG Pathway and Identification of Novel Neuromodulatory Agents Using Drosophila Melanogaster.
Creator: Caplan, Stacee Lee, Dawson-Scully, Ken, Milton, Sarah L., Florida Atlantic University, Charles E. Schmidt College of Science, Department of Biological Sciences
Abstract/Description: Oxidant stress and injury is inherent in many human diseases such as ischemic vascular and respiratory diseases, heart failure, myocardial infarction, stroke, perinatal and placental insufficiencies, diabetes, cancer, and numerous psychiatric and neurodegenerative disorders. Finding novel therapeutics to combat the deleterious effects of oxidative stress is critical to create better therapeutic strategies for many conditions that have few treatment options. This study used the anoxia-tolerant...
Show moreOxidant stress and injury is inherent in many human diseases such as ischemic vascular and respiratory diseases, heart failure, myocardial infarction, stroke, perinatal and placental insufficiencies, diabetes, cancer, and numerous psychiatric and neurodegenerative disorders. Finding novel therapeutics to combat the deleterious effects of oxidative stress is critical to create better therapeutic strategies for many conditions that have few treatment options. This study used the anoxia-tolerant fruit fly, Drosophila melanogaster, to investigate endogenous cellular protection mechanisms and potential interactions to determine their ability to regulate synaptic functional tolerance and cell survival during acute oxidative stress. The Drosophila larval neuromuscular junction (NMJ) was used to analyze synaptic transmission and specific motor axon contributions. Drosophila Schneider 2 (S2) cells were used to assess viability. Acute oxidative stress was induced using p harmacological paradigms that generate physiologically relevant oxidant species: mitochondrial superoxide production induced by sodium azide (NaN3) and hydroxyl radical formation via hydrogen peroxide (H2O2). A combination of genetic and pharmacological approaches were used to explore the hypothesis that endogenous protection mechanisms control cellular responses to stress by manipulating ion channel conductance and neurotransmission. Furthermore, this study analyzed a group of marine natural products, pseudopterosins, to identify compounds capable of modulating synaptic transmission during acute oxidative stress and potential novel neuromodulatory agents.
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Date Issued: 2015
PURL: http://purl.flvc.org/fau/fd/FA00004487, http://purl.flvc.org/fau/fd/FA00004487
Subject Headings: Drosophila melanogaster -- Life cycles, Oxidative stress -- Ecophysiology, Oxidative stress -- Prevention, Protein kinases, Proteins -- Chemical modification
Format: Document (PDF)

Title: Characterization of receptor protein tyrosine phosphatase PTP69D in the giant fiber circuit.
Creator: Lee, LaTasha Hoskins, Godenschwege, Tanja A., Florida Atlantic University, Charles E. Schmidt College of Science, Department of Biological Sciences
Abstract/Description: PTP69D is a receptor protein tyrosine phosphatase (RPTP) with two intracellular catalytic domains (Cat1 and Cat2), which has been shown to play a role in axon outgrowth and guidance of embryonic motorneurons, as well as targeting of photoreceptor neurons in the visual system of Drosophila melanogaster. Here, we characterized the developmental role of PTP69D in the giant fiber (GF) neurons; two interneurons in the central nervous system (CNS) that control the escape response of the fly. In...
Show morePTP69D is a receptor protein tyrosine phosphatase (RPTP) with two intracellular catalytic domains (Cat1 and Cat2), which has been shown to play a role in axon outgrowth and guidance of embryonic motorneurons, as well as targeting of photoreceptor neurons in the visual system of Drosophila melanogaster. Here, we characterized the developmental role of PTP69D in the giant fiber (GF) neurons; two interneurons in the central nervous system (CNS) that control the escape response of the fly. In addition to guidance and targeting functions, our studies reveal an additional role for PTP69D in synaptic terminal growth in the CNS. We found that inhibition of phosphatase activity in catalytic domain (Cat1) proximal to the transmembrane domain did not affect axon guidance or targeting but resulted in stunted terminal growth of the GFs. Cell autonomous rescue and knockdown experiments demonstrated a function for PTP69D in the GFs, but not its postsynaptic target neurons. In addition,complementation studies and structure-function analyses revealed that for GF terminal growth, Cat1 function of PTP69D requires the immunoglobulin and the Cat2 domain but not the fibronectin type III repeats nor the membrane proximal region. In contrast, the fibronectin type III repeats, but not the immunoglobulin domains, were previously shown to be essential for axon targeting of photoreceptor neurons. Thus, our studies uncover a novel role for PTP69D in synaptic terminal growth in the CNS that is mechanistically distinct from its function during earlier developmental processes.
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Date Issued: 2014
PURL: http://purl.flvc.org/fau/fd/FA00004301, http://purl.flvc.org/fau/fd/FA00004301
Subject Headings: Drosophila melanogaster., Protein-tyrosine phosphatase--Metabolism., Protein-tyrosine kinase., Protein kinases--Inhibitors., Phosphoprotein phosphatases., Transcription factors., Cell receptors., Cellular signal transduction.
Format: Document (PDF)

Title: Purification and characterization of two members of the protein tyrosine phosphatase family: dual specificity phosphatase PVP and low molecular weight phosphatase WZB.
Creator: Livingston, Paula A., Charles E. Schmidt College of Science, Department of Chemistry and Biochemistry
Abstract/Description: Two protein tyrosine phosphatases, dual specificity phosphatase PVP and low molecular weight phosphatase WZB were purified and characterized. PVP was expressed as inclusion bodies and a suitable purification and refolding method was devised. Enzyme kinetics revealed that p-nitrophenylphosphate and (Sb(B-naphthyl phosphate were substrates with KM of 4.0mM and 8.1mM respectively. PVP showed no reactivity towards phosphoserine. Kinetic characterization of WZB showed that only...
Show moreTwo protein tyrosine phosphatases, dual specificity phosphatase PVP and low molecular weight phosphatase WZB were purified and characterized. PVP was expressed as inclusion bodies and a suitable purification and refolding method was devised. Enzyme kinetics revealed that p-nitrophenylphosphate and (Sb(B-naphthyl phosphate were substrates with KM of 4.0mM and 8.1mM respectively. PVP showed no reactivity towards phosphoserine. Kinetic characterization of WZB showed that only pnitrophenylphosphate was a substrate with no affinity for Ç-naphthyl phosphate and phosphoserine. Optimal conditions for activity with PNPP were found at a pH of 5 with a KM of 1.1mM, kcat of 35.4s-1 and kcat/KM of 32.2s-1mM-1. Inhibition studies showed that phosphate, fluoride, and molybdate were competitive inhibitors with Ki of 3.2mM, 71.7mM, and 50.4(So(BM respectively and hydrogen peroxide abolished activity. Active site mutants of WZB Cys9Ser and Asp115Asn showed no activity.
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Date Issued: 2009
PURL: http://purl.flvc.org/FAU/332911
Subject Headings: Protein-tyrosine phosphatase, Cellular signal transduction, Cell cycle, Regulation, Membrane proteins, Structure-activity relationships, Protein kinases
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.
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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: Determining the subcellular localization of a group II p21-activated kinase - PAK6.
Creator: John, Ciny, Charles E. Schmidt College of Medicine, Department of Biomedical Science
Abstract/Description: p-21-activated kinase 6 (PAK6) is a serine-threonine protein kinase originally identified as an Androgen Receptor (AR) interacting protein. In current study, we determined the subcellular localization of PAK6 through mutational analysis. We have found that the N-terminal CRIB domain is partly responsible for plasma membrane targeting, the region between amino acid residues #292 to #368 is functionally relevant to plasma membrane localization and that amino acid residues #119 through #190 are...
Show morep-21-activated kinase 6 (PAK6) is a serine-threonine protein kinase originally identified as an Androgen Receptor (AR) interacting protein. In current study, we determined the subcellular localization of PAK6 through mutational analysis. We have found that the N-terminal CRIB domain is partly responsible for plasma membrane targeting, the region between amino acid residues #292 to #368 is functionally relevant to plasma membrane localization and that amino acid residues #119 through #190 are responsible for nuclear targeting of PAK6, in addition to a stretch of positively charged N-terminal residues (#2-#11) since mutants lacking this sequence mis-localizes to cytoplasm. In junction forming epithelial cells, PAK6 is demonstrated to co-localize with B-catenin at adherens junctions, suggesting that PAK6 is an activation-dependent event and that PAK6 translocates from plasma membrane to the cytoplasm in response activation via the PKA signal pathway.
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Date Issued: 2012
PURL: http://purl.flvc.org/FAU/3355569
Subject Headings: Cellular signal transduction, Serine proteinases, Phosphorylation, Protein kinases, Pathophysiology, Phosphoroproteins, Metabolism
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.
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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: Temporal response of creatine kinase and fibroblast growth factor-21 to high and low repetition resistance training programs.
Creator: Blanco, Rocky, Zourdos, Michael C., Florida Atlantic University, College of Education, Department of Exercise Science and Health Promotion
Abstract/Description: The purpose of this study was to examine the acute and temporal response of CK- MM and FGF-21 to 3-day/wk. different repetition-range, volume-equated resistance training programs over 8-weeks in previously trained males. Sixteen trained, college- aged males were counterbalanced into high (DUP-HR) or low (DUP-LR) repetition groups. Subjects performed the squat and bench press 3x/wk. for 8 weeks. Blood samples were collected at various intervals throughout the study. Trained individuals did not...
Show moreThe purpose of this study was to examine the acute and temporal response of CK- MM and FGF-21 to 3-day/wk. different repetition-range, volume-equated resistance training programs over 8-weeks in previously trained males. Sixteen trained, college- aged males were counterbalanced into high (DUP-HR) or low (DUP-LR) repetition groups. Subjects performed the squat and bench press 3x/wk. for 8 weeks. Blood samples were collected at various intervals throughout the study. Trained individuals did not elicit significant acute or chronic changes in CK-MM or FGF-21 following training and the lack of change was present in both groups. Additionally, neither biomarker correlated with changes in 1RM strength. There was a very strong correlation between acute mean (r=0.95) and acute percentage change (r=0.97) increase from pre training to post training in week #1. Additionally, a moderate correlation in percentage change was observed (r=0.59) of both biomarkers from pre training to 48 hours post training in week #2.
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Date Issued: 2015
PURL: http://purl.flvc.org/fau/fd/FA00004429, http://purl.flvc.org/fau/fd/FA00004429
Subject Headings: Bioenergetics, Cellular signal transduction, Fibroblast growth factors, Metabolic syndrome -- Pathophysiology, Protein kinases -- Inhibitors -- Therapeutic use
Format: Document (PDF)

FAU Theses and Dissertations (9)	+ -
HBOI Faculty Contributions (2)	+ -
FAU Graduate Student Research (1)	+ -

Protein kinases (5)	+ -
Cellular signal transduction (3)	+ -
Cyclic GMP-Dependent Protein Kinases (3)	+ -
Drosophila melanogaster (3)	+ -
Protein kinase C (2)	+ -

Ascidians (1)	+ -
Bioenergetics (1)	+ -
Caenorhabditis elegans (1)	+ -
Carbazoles (1)	+ -
Cell cycle (1)	+ -
Cell receptors. (1)	+ -
Cellular signal transduction. (1)	+ -
Colorimetric analysis (1)	+ -
Cytotoxins (1)	+ -
Diglycerides (1)	+ -
Drosophila melanogaster -- Life cycles (1)	+ -
Drosophila melanogaster. (1)	+ -
Ecophysiology (1)	+ -
Fibroblast growth factors (1)	+ -
Heat shock proteins (1)	+ -
Hyperthermia (1)	+ -
Indoles (1)	+ -
Life cycles (1)	+ -
Membrane proteins (1)	+ -
Metabolic syndrome -- Pathophysiology (1)	+ -
Metabolism (1)	+ -
Neuroprotection (1)	+ -
Neuroprotective agents (1)	+ -
Oxidative Stress (1)	+ -
Oxidative stress -- Ecophysiology (1)	+ -

Electronic Thesis or Dissertation (9)	+ -
article (2)	+ -
abstract (1)	+ -

Charles E. Schmidt College of Science (7)	+ -
Department of Biological Sciences (6)	+ -
Florida Atlantic University (6)	+ -
Dawson-Scully, Ken (5)	+ -
Milton, Sarah L. (3)	+ -

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