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An immunological approach to the study of the tumorous-head trait in Drosophila melanogaster
Title
An immunological approach to the study of the tumorous-head trait in Drosophila melanogaster.
Creator
Weihe, Patricia Neuhaus, null, null, Social Sciences
Abstract / Description
Florida Technological University College of Natural Sciences Thesis;
Date Issued
1975
Identifier
CFR0004337, ucf:52990
Format
Document (PDF)
PURL
http://purl.flvc.org/ucf/fd/CFR0004337
STUDIES OF NORSPERMIDINE UPTAKE IN DROSOPHILA SUGGEST THE EXISTENCE OF MULTIPLE POLYAMINE TRANSPORT PATHWAYS
Title
STUDIES OF NORSPERMIDINE UPTAKE IN DROSOPHILA SUGGEST THE EXISTENCE OF MULTIPLE POLYAMINE TRANSPORT PATHWAYS.
Creator
Dieffenbach, Michael, Von Kalm, Laurence, Teter, Kenneth, University of Central Florida
Abstract / Description
Polyamines are a class of essential nutrients involved in many basic cellular processes such as gene expression, cell proliferation, and apoptosis. Without polyamines, cell growth is delayed or halted. Cancerous cells require an abundance of polyamines through a combination of synthesis and transport from the extracellular environment. An FDA-approved drug, D,L-?-difluoromethylornithine (DFMO), blocks polyamine synthesis but is ineffective at inhibiting cell growth due to polyamine transport....
Show morePolyamines are a class of essential nutrients involved in many basic cellular processes such as gene expression, cell proliferation, and apoptosis. Without polyamines, cell growth is delayed or halted. Cancerous cells require an abundance of polyamines through a combination of synthesis and transport from the extracellular environment. An FDA-approved drug, D,L-?-difluoromethylornithine (DFMO), blocks polyamine synthesis but is ineffective at inhibiting cell growth due to polyamine transport. Thus, there is a need to develop drugs that inhibit polyamine transport to use in combination with DFMO. Surprisingly, little is known about the polyamine transport system in humans and other eukaryotes. Understanding the transport system would allow us to identify compounds that inhibit polyamine transport, which could then be used in tandem with DFMO to treat cancer. Our laboratory has identified one gene in Drosophila, called CG32000, as a component of this transport system, and numerous other candidate genes remain to be tested. To better characterize this system, this project investigated the ability of the Drosophila transport system to take up a toxic polyamine analogue called norspermidine, with the initial goal of developing a new screening method to find polyamine transport genes. My experiments have demonstrated significant differences in norspermidine uptake and toxicity between C. elegans and Drosophila which may imply a secondary polyamine transport system in higher eukaryotes. In the long term, it is hoped that this thesis will facilitate the development of more effective cancer medications by providing new information about the polyamine transport system.
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Date Issued
2018
Identifier
CFH2000294, ucf:45869
Format
Document (PDF)
PURL
http://purl.flvc.org/ucf/fd/CFH2000294
GEOGRAPHIC VARIATION IN POST-MATING IMMUNE GENE EXPRESSION INDROSOPHILA MELANOGASTER
Title
GEOGRAPHIC VARIATION IN POST-MATING IMMUNE GENE EXPRESSION INDROSOPHILA MELANOGASTER.
Creator
Pinzone, Cheryl, Fedorka, Kenneth, University of Central Florida
Abstract / Description
An organism's immune response may vary due to pathogen pressure in its environment, as well as due to interactions with other organisms. These factors, along with geographic rules (i.e. Gloger's rule) may influence the geographic distribution of the immune response within populations of a species. Here we use real-time quantitative PCR to measure the immune gene expression in six populations collected along the eastern U.S. of Drosophila melanogaster after mating. Antimicrobial genes...
Show moreAn organism's immune response may vary due to pathogen pressure in its environment, as well as due to interactions with other organisms. These factors, along with geographic rules (i.e. Gloger's rule) may influence the geographic distribution of the immune response within populations of a species. Here we use real-time quantitative PCR to measure the immune gene expression in six populations collected along the eastern U.S. of Drosophila melanogaster after mating. Antimicrobial genes did not show significant differences in expression due to location, whereas we did observe differences in anti-fungal and pro-phenoloxidase (anti-macromolecule) related genes. These differences in anti-macromolecule resistance are correlated with the latitude of the population opposite of which we would expect by Gloger's rule. We also determined that males and females from different populations tended to drive the differences we detected. Taken together, these results suggest that geographic factors influence genes involved in fungal and macro-pathogens defense post-mating.
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Date Issued
2010
Identifier
CFE0003159, ucf:48617
Format
Document (PDF)
PURL
http://purl.flvc.org/ucf/fd/CFE0003159
Climate Change and the Evolution of Insect Immune Function
Title
Climate Change and the Evolution of Insect Immune Function.
Creator
Perry, Danae, Fedorka, Kenneth, Jenkins, David, Hoffman, Eric, University of Central Florida
Abstract / Description
Insects are ectothermic organisms that have physiological, behavioral and life-history traits directly influenced by their environment. Investigations have shown that many insects use melanin to permanently darken their cuticles in cooler or drier climates to improve thermoregulation and desiccation resistance. Melanin is a major component of pathogen defense in insects. This suggests that environmentally driven adaptive changes in cuticular melanin may non-adaptively shape insect immune...
Show moreInsects are ectothermic organisms that have physiological, behavioral and life-history traits directly influenced by their environment. Investigations have shown that many insects use melanin to permanently darken their cuticles in cooler or drier climates to improve thermoregulation and desiccation resistance. Melanin is a major component of pathogen defense in insects. This suggests that environmentally driven adaptive changes in cuticular melanin may non-adaptively shape insect immune function. This hypothesis has been referred to as climate-related Cuticle Dependent Immune Investment (climate-related CDII). Climate-related CDII also suggests that a warming climate could lead to the evolution of a weakened melanin-based immune response due to direct selection for lighter cuticles. Climate-related CDII has not been investigated with regard to climate change. Using Drosophila melanogaster, the first part of this study investigated if the documented pattern of lowered immune function in warmer temperatures offsets the expected gain in metabolic rate. The second part of this project investigated how a warming thermal environment will affect the evolution of insect immune function by quantifying changes in melanization and immune function over multiple generations in a changing thermal environment. In the first investigation there was evidence for weakened immune function in males, while females saw an offset by gaining a metabolic boost. The second investigation showed evidence that warming treatments evolved lowered overall immune function. This project gives evidence that insect immune function has the potential to be weakened by increasing temperatures. Insect immune function is a major contributing factor to insect abundances. A decrease in beneficial insects or an increase in harmful insects or pathogens they vector could have detrimental environment and human health consequences.?
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Date Issued
2017
Identifier
CFE0006638, ucf:51256
Format
Document (PDF)
PURL
http://purl.flvc.org/ucf/fd/CFE0006638
NOTOPLEURAL MUTATIONS ENHANCE DEFECTS IN IMAGINAL DISC EPITHELIAL MORPHOGENESIS AND MACROCHETE ELONGATION ASSOCIATED WITH MUTATIONS IN THE STUBBLE-STUBBLOID LOCUS
Title
NOTOPLEURAL MUTATIONS ENHANCE DEFECTS IN IMAGINAL DISC EPITHELIAL MORPHOGENESIS AND MACROCHETE ELONGATION ASSOCIATED WITH MUTATIONS IN THE STUBBLE-STUBBLOID LOCUS.
Creator
Ruggiero, Robert, von Kalm, Laurence, University of Central Florida
Abstract / Description
The Stubble-stubbloid locus encodes a transmembrane serine protease (Stubble) necessary for the proper formation of sensory bristles, and the morphogenesis of leg and wing epithelia. Genetic and cell biological analysis indicate a role for Stubble in actin cytoskeletal dynamics and cell shape changes in developing epithelia and bristles. Previously reported genetic interactions between Stubble and the Rho1 signaling pathway suggest Stubble influences actin cytoskeleton dynamics in developing...
Show moreThe Stubble-stubbloid locus encodes a transmembrane serine protease (Stubble) necessary for the proper formation of sensory bristles, and the morphogenesis of leg and wing epithelia. Genetic and cell biological analysis indicate a role for Stubble in actin cytoskeletal dynamics and cell shape changes in developing epithelia and bristles. Previously reported genetic interactions between Stubble and the Rho1 signaling pathway suggest Stubble influences actin cytoskeleton dynamics in developing imaginal discs through interactions with the Rho1 pathway. This work will discuss a genetic screen conducted to further investigate the role of Stubble in bristle and imaginal disc morphogenesis. From 50,000 EMS-mutagenized chromosomes 12 enhancers of the recessive sbd201 allele were identified, including 6 new sbd alleles. Consistent with the current understanding of genetic interactions regulating imaginal disc morphogenesis, mutations in two Rho1 pathway genes, zipper (2 alleles) and Rho1, were isolated. Additionally, three new mutant enhancers of sbd201 were isolated, one of which has been identified as an allele of the cadherin gene Dacshous, another as an allele of the muscle myosin heavy chain gene, and the last as an allele of Notopleural (Np). Dominant and recessive mutations in the Stubble locus interact with the Np allele identified in this screen, in regards to both limb and bristle development, respectively. Mutations in the Np locus were first identified in 1936, but this locus remains poorly characterized and has never been cloned The genetic and phenotypic characterization of Np will be discussed along with experiments that have mapped the position of the Np locus to a 50kb region at the border of the 44F12, 45A1 cytological regions.
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Date Issued
2006
Identifier
CFE0001347, ucf:46986
Format
Document (PDF)
PURL
http://purl.flvc.org/ucf/fd/CFE0001347
A chemical and genetic approach to study the polyamine transport system in Drosophila
Title
A chemical and genetic approach to study the polyamine transport system in Drosophila.
Creator
Wang, Minpei, Vonkalm, Laurence, Phanstiel, Otto, Teter, Kenneth, Ballantyne, John, University of Central Florida
Abstract / Description
Polyamines are small cationic molecules that play important roles in most vital cellular processes including cell growth and proliferation, regulation of chromatin structure, translation and programmed cell death. Cellular polyamine pools are maintained by a balance between biosynthesis and transport (export and import). Increased polyamine biosynthesis activity and an active transport system are characteristics of many cancer cell lines, and polyamine depletion has been shown to be a viable...
Show morePolyamines are small cationic molecules that play important roles in most vital cellular processes including cell growth and proliferation, regulation of chromatin structure, translation and programmed cell death. Cellular polyamine pools are maintained by a balance between biosynthesis and transport (export and import). Increased polyamine biosynthesis activity and an active transport system are characteristics of many cancer cell lines, and polyamine depletion has been shown to be a viable anticancer strategy. Polyamine levels can be depleted by ?-difluoromethylornithine (DFMO), an inhibitor of the key polyamine biosynthesis enzyme ornithine decarboxylase. However, malignant cells often circumvent DFMO therapy by up-regulating polyamine import; therefore, there is a need to develop compounds that inhibit polyamine transport. Collectively, DFMO and polyamine transport inhibitors provide the basis for a combination therapy leading to effective intracellular polyamine depletion. Using a Drosophila leg imaginal disc model for polyamine transport, I studied three candidate transport inhibitors (Ant444, Trimer44 and Triamide44) for their ability to inhibit transport in the Drosophila model. Ant444 and Trimer44 effectively inhibited the uptake of the toxic polyamine analog Ant44 that gains entry to cells via the polyamine transport system. Ant444 and Trimer44 were also able to inhibit the import of exogenous polyamines into DFMO-treated imaginal discs. Triamide44 was an ineffective inhibitor, however a structurally redesigned compound, Triamide444, showed a 50-fold increase in transport inhibition and was comparable to Ant444 and Trimer44. Ant444 and Trimer44 showed differences in their relative abilities to block import of specific polyamines, and I therefore asked if a cocktail of these inhibitors would be more effective than either alone. My data show that a cocktail of polyamine transport inhibitors is more effective than single inhibitors when used in combination with DFMO, and suggests the existence of multiple polyamine transport systems. To further the development of effective transport inhibitors it is important to identify components of the transport system. The mechanism of polyamine transport in multicellular organisms including mammals is still unknown. Our laboratory has developed a simple assay to detect components of the transport system using RNAi knockdown and over-expression of candidate genes. However, the assay requires that animals live until the pupal stage of development. Pleiotropic effects of individual gene products following over-expression or knockdown may result in early developmental lethality for reasons unrelated to polyamine transport. Our assay is based on the GAL4/UAS system and involves the use of enhancers driving GAL4 expression (GAL4 driver). GAL4 in turn determines the expression level of UAS-candidate gene constructs (UAS responder). I reasoned that in some cases it might be possible to bypass early lethality by judicious choice of drivers that reduce responder expression, thus permitting survival to the pupal phase. To this end, I used five imaginal disc drivers (30A, 71B, 32B, 69B, and T80) as well as a ubiquitously expressed control driver to over-express and knockdown EGFR and components of the Rho signaling pathway. The relative strength of each driver was ranked, and I was able to demonstrate in principle that animals could survive to later stages of development in a manner that correlated with the relative strength of the driver. The approach I developed is broadly applicable to other studies of Drosophila development.To identify new components of the polyamine transport system I studied the role of proteoglycans in this process. The proteoglycan glypican-1 has been previously implicated in mammalian polyamine transport. In particular, the heparin sulfate side chains of glypican-1 appear to play an important role. In order to extend our knowledge of the role of proteoglycans in polyamine transport, I examined the role of the core proteoglycans perlecan and syndecan as well as genes encoding enzymes in the heparin sulfate and chondroitin sulfate biosynthetic pathways. I was able to confirm a role for glypican-1 in polyamine transport in imaginal discs but not in whole animals. This may indicate that glypican-1 is not required for polyamine uptake through the gut. Studies of genes encoding perlecan, syndecan and enzymes in the heparin sulfate and chondroitin sulfate biosynthetic pathways did not reveal a role for these genes in polyamine transport. These studies were conducted in whole animals and my data may reflect tissue-specific differences between the imaginal disc and gut transport systems where transport in imaginal discs is proteoglycan dependent and transport in the gut is not.
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Date Issued
2017
Identifier
CFE0007297, ucf:52162
Format
Document (PDF)
PURL
http://purl.flvc.org/ucf/fd/CFE0007297
MAPPING AND CHARACTERIZATION OF 18-5 AND 12-5, GENES WHICH POTENTIALLY LINK THE RHOA SIGNALING PATHWAY TO THE ECDYSONE RESPONSE IN DROSOPHILA EPITHELIAL MORPHOGENESIS.
Title
MAPPING AND CHARACTERIZATION OF 18-5 AND 12-5, GENES WHICH POTENTIALLY LINK THE RHOA SIGNALING PATHWAY TO THE ECDYSONE RESPONSE IN DROSOPHILA EPITHELIAL MORPHOGENESIS.
Creator
Fox, Samuel, von Kalm, Laurence, University of Central Florida
Abstract / Description
Systemic steroid hormone and intracellular signaling pathways are known to act cooperatively during the development of vertebrate and invertebrate epithelia. However, the mechanism of this interaction is poorly understood. Morphogenesis of Drosophila leg imaginal disc epithelia is regulated both by the steroid hormone 20-hydroxyecdysone (ecdysone) and the RhoA GTPase signaling pathway. Recent evidence suggests that these pathways act cooperatively to control imaginal disc morphogenesis. Thus,...
Show moreSystemic steroid hormone and intracellular signaling pathways are known to act cooperatively during the development of vertebrate and invertebrate epithelia. However, the mechanism of this interaction is poorly understood. Morphogenesis of Drosophila leg imaginal disc epithelia is regulated both by the steroid hormone 20-hydroxyecdysone (ecdysone) and the RhoA GTPase signaling pathway. Recent evidence suggests that these pathways act cooperatively to control imaginal disc morphogenesis. Thus, leg imaginal disc morphogenesis is an excellent system in which to study the interaction of steroid hormone and intracellular signaling pathways. We have identified mutations in three genes, 12-5, 18-5, and 31-6, with roles in the morphogenesis of leg epithelia. Of particular interest, these mutations interact genetically with each other, mutations in the RhoA signaling pathway, and the ecdysone regulated Sb-sbd (Stubble) transmembrane serine protease. This suggests that the 12-5, 18-5, and 31-6 gene products may link hormone and RhoA signaling responses. The goal of this research was to identify and characterize the 18-5 and 12-5 genes in order to discern the mechanistic relationship between the RhoA pathway and ecdysone hierarchy.18-5 and 12-5 were precisely mapped to molecular locations within the Drosophila genome utilizing a P-element recombination mapping technique. This work narrowed the location of the 18-5 locus to within an interval of 112 kb within the Drosophila genome sequence. This interval contains 17 known and predicted genes. I also mapped the location of the 12-5 locus to a 2.6 Mb interval of the 2nd chromosome. Based on phenotypic analyses and the site of the molecularly mapped interval, a candidate gene for the 18-5 mutation was identified. Sequence analysis of the candidate gene was inconclusive and requires further analysis. Genetic interaction assays indicate that the 18-5 gene product acts upstream or at the level of Rho kinase in the RhoA signaling pathway.
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Date Issued
2006
Identifier
CFE0001290, ucf:46882
Format
Document (PDF)
PURL
http://purl.flvc.org/ucf/fd/CFE0001290
The Role of the Y-Chromosome in the Evolution of Autosomally Coded Traits
Title
The Role of the Y-Chromosome in the Evolution of Autosomally Coded Traits.
Creator
Kutch, Ian, Fedorka, Kenneth, Vonkalm, Laurence, Hoffman, Eric, University of Central Florida
Abstract / Description
Recent work indicates that the Y-chromosome of the fruit fly Drosophila melanogaster can influence gene regulation on the autosomes and X chromosome. This newly discovered function of the Y has the potential to dramatically shape the regulatory evolution of numerous genes that reside throughout the genome; even for genes that code for both male and female traits. Given that the mechanism underlying the Y-linked influence on gene expression in D. melanogaster appears to exist in other...
Show moreRecent work indicates that the Y-chromosome of the fruit fly Drosophila melanogaster can influence gene regulation on the autosomes and X chromosome. This newly discovered function of the Y has the potential to dramatically shape the regulatory evolution of numerous genes that reside throughout the genome; even for genes that code for both male and female traits. Given that the mechanism underlying the Y-linked influence on gene expression in D. melanogaster appears to exist in other independently evolved heterogametic sex chromosomes, the evolutionary implications of Y-linked regulatory variation (YRV) deserves to be explored. These implications include the potential for Y-chromosomes to facilitate the adaptive evolution of sexually dimorphic gene expression, and the potential for the Y to constrain evolutionary rates in both males and females (depending on the nature of the YRV effect). Unfortunately, the evolutionary implications of this potentially widespread and significant phenomenon have yet to be explored. My dissertation addresses this knowledge gap by determining the influence YRV has on the evolution of autosomally coded traits in D. melanogaster. First, we address the potential for selection to shape YRV by determining if YRV (i) exists within natural populations (i.e. where natural selection operates), and (ii) has any influence on male fitness-related autosomal traits. Second, we address if YRV can facilitate the adaptive evolution of sexually dimorphic gene expression by testing for the presence of Y-linked additive genetic variation. To this end, we investigate the physiological properties of select Y-chromosomes across multiple genetic backgrounds. Third, we address if YRV can constrain adaptive evolution for autosomally coded traits by employing artificial selection on replicate populations that contain either multiple Y-chromosomes (i.e. contain YRV) or only a single Y-chromosome (no YRV). The following studies present evidence that YRV does exist within populations where natural selection operates. We show significant levels of YRV on X-linked and autosomal immune gene expression in wild caught D. melanogaster from a single natural population. Furthermore, YRV effects on immune related genes show a significantly positive correlation to a male fly's ability to fight an immune challenge (an important aspect of organismal fitness). Estimated physiological properties of YRV support previous interpopulation studies showing strong non-additive effect dependent on the autosomal genetic background with which Y-chromosome's are paired with. Physiological epistasis can manifest as additive genetic variation on a population level, but our experimental evolution study suggest that YRV constrains rather than facilitates the evolution of the autosomal coded geotaxis behavior. Ultimately, this dissertation provides evidence that YRV has the potential to influence how autosomal traits evolve and that population level studies of YRV indicate a potential constraint to the adaptive evolution of autosomal traits. If these trends are common and YRV is a wide spread phenomenon, Y-chromosomes have the potential to influence how autosomal traits evolve.
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Date Issued
2017
Identifier
CFE0006756, ucf:51873
Format
Document (PDF)
PURL
http://purl.flvc.org/ucf/fd/CFE0006756