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The Effects of Inbreeding on Fitness Traits in the Critically Endangered Attwater’s Prairie-chicken

Description: The goals of captive breeding programs for endangered species include preserving genetic diversity and avoiding inbreeding. Typically this is accomplished by minimizing population mean kinship; however, this approach becomes less effective when errors in the pedigree exist and may result in inbreeding depression, or reduced survival. Here, both pedigree- and DNA-based methods were used to assess inbreeding depression in the critically endangered Attwater’s prairie-chicken (Tympanuchus cupido attwateri). Less variation in the pedigree-based inbreeding coefficients and parental relatedness values were observed compared to DNA-based measures suggesting that errors exist in the pedigree. Further, chicks identified with high parental DNA-based relatedness exhibited decreased survival at both 14- and 50-days post-hatch. A similar pattern was observed in later life stages (> 50 days post-hatch) with birds released to the wild; however, the pattern varied depending on the time post-release. While DNA-based inbreeding coefficient was positively correlated with mortality to one month post-release, an opposite pattern was observed at nine months suggesting purging of deleterious alleles. I also investigated whether immunocompetence, or the ability to produce a normal immune response, was correlated with survival; however, no significant correlation was observed suggesting that inbreeding was a more important factor influencing survival. Pairing individuals for breeding by minimizing DNA-based parental relatedness values resulted in a significant increase in chick survival. This study highlights the importance of using DNA-based methods to avoid inbreeding depression when errors exist in the pedigree.
Date: August 2014
Creator: Hammerly, Susan C.
Partner: UNT Libraries

Effects of Methanol, Atrazine, and Copper on the Ultrastructure of Pseudokirchneriella Subcapitata (Selenastrum Capricornutum).

Description: The toxicity of methanol, atrazine, and copper to Pseudokirchneriella subcapitata (Korshikov) Hindák historically referred to as Selenastrum capricornutum Printz were determined following 96 hrs growth in a modified Goram's growth media. Methanol and atrazine inhibited fluorescence readings in the cultures by 50% (IC50) at concentrations of 2% and 82 µg/l respectively. These toxicity values compared favorably to other published reports. The IC50 for copper was 160 µg/l which is substantially higher than reported values. This is understandable because of the high chelating capacity of Goram's media. The use of stereologically derived relative volume in the chloroplasts, mitochondria, lipid bodies, phosphate bodies, and nucleus was investigated to determine if it could be used as a sensitive endpoint in toxicity tests. The volume fractions for the chloroplasts and mitochondria were normally distributed in control cells while the nuclei, phosphate bodies, and lipid bodies were not. The chloroplasts were the most dominate organelle occupying a mean relative volume of 46% and mitochondria occupied a mean relative volume of 3%. The nucleus and phosphate bodies occupied a median relative volume of 7% and 2% respectively. The lipid bodies were rare in section profile and no meaningful median relative volume could be calculated. Up to the 82nd percentile of sectioned profiles contained no recognizable lipid bodies. The use of relative volume was not a sensitive endpoint for use in toxicity tests. No significant differences in relative volume could be detected in the nucleus or phosphate bodies following any treatment. Limited differences were detected in the mitochondria, chloroplasts, and lipid bodies. The only significant differences that appear to be biologically significant occurred in methanol treated cells where an increase in the lipid bodies' relative volume was apparently concentration dependent. Significant differences in the relative volume of mitochondria and chloroplasts do not appear to be biologically significant.
Date: May 2004
Creator: Garrett, David C.
Partner: UNT Libraries

The Effects of Organic Surface Amendments on Soil Nutrients and Initial Tree Establishment

Description: This study examined the effects of replicating woodland soil surface horizonation on the nutrient status of underlying soils and the initial establishment and growth of trees. A total of 283 container grown trees were planted in a bufferzone around a future landfill site. Control amendments consisted of an 8 cm layer (0.5 m3) of wood chips applied in a circular area of 4.6 m2 around the trees' planting pit. For the treatment, a 2.5 cm layer of composted biosolids (0.15 m3 or 80 Mg/ha) was applied in a circular area of 4.6 m2 around the trees' planting pit followed by an 8 cm layer (0.5 m3) of wood chips. The results indicate that the replication of woodland soil surface attributes using composted biosolids can significantly improve the nutrient status of underlying soil. Some significant effects were seen under control conditions, too. However, the effects on tree establishment and growth parameters were, for the most part, not statistically significant.
Date: May 1999
Creator: Thuesen, Kevin (Kevin Andrew)
Partner: UNT Libraries

Effects of Peripheral Nerve Injury on the Cells of the Dorsal Root Ganglion: a Role for Primary Cilia

Description: Primary cilia are ubiquitous sensory organelles found on most cell types including cells of the dorsal root ganglia (DRG). The DRG are groups of peripheral neurons that relay sensory information from the periphery to the CNS. Other cell types in the DRG include a type of glial cell, the satellite glial cells (SGCs). The SGCs surround the DRG neurons and, with the neurons, form functional sensory units. Currently are no reports describing the numbers of DRG cells that have cilia. We found that 26% of the SGCs had primary cilia. The incidence of cilia on neurons varied with neuron size, a property that roughly correlates with physiological characteristics. We found that 29% of the small, 16% of the medium and 5% of the large neurons had primary cilia. Primary cilia have been shown to have a role in cell proliferation in a variety of cell types. In some of the cells the cilia mediate the proliferative effects of Sonic hedgehog (Shh). In the CNS, Shh signaling through primary cilia affects proliferation during development as well as following injury, but no studies have looked at this function in the PNS. The SGCs and neurons of the DRG undergo complex changes following peripheral nerve injury such as axotomy. One marked change seen after axotomy is SGC proliferation and at later stages, neuronal death. We found that following axotomy there is a significant increase in the percentage of SGCs with primary cilia. We also found a significant increase in the percentage of medium-sized neurons with primary cilia. In other experiments we tested the idea that Shh plays a role in SGC proliferation. When Shh signaling was blocked following axotomy we found decreased proliferation of SGCs. This is the first report of a change in the percentage of cells with cilia following injury in ...
Date: December 2012
Creator: Smith, Sarah K.
Partner: UNT Libraries

Effects of Triclosan, Triclocarban, and Caffeine Exposure on the Development of Amphibian Larvae.

Description: Triclosan and triclocarban are antimicrobials found in numerous consumer products, while caffeine is the most commonly consumed stimulant by humans. This study was undertaken to determine the effects of triclosan, triclocarban, and caffeine on the development and physiology of amphibian larvae. LC50 values of triclosan and triclocarban were determined after 96 hours for three North American larval species: Acris crepitans blanchardii, Bufo woodhousii woodhousii, Rana sphenocephala, and for a common amphibian developmental model: Xenopus laevis. Amphibian larvae were most sensitive to triclosan and triclocarban exposure during early development based upon 96-hour LC50 values. Heart rates for X. laevis and North American larvae exposed to triclosan were variable throughout development. However, significantly lower heart rates were observed in all larvae exposed to triclocarban. Metabolic rates of X. laevis and R. sphenocephala larvae exposed to triclosan were significantly affected in larvae exposed to ½ LC50 and the LC50 concentration. Metabolic rates of X. laevis larvae exposed to triclocarban were significantly affected by exposure to ½ LC50 concentrations in three of four stages investigated. No significant differences were observed in North American larvae exposed to triclocarban. Tissue uptake, lipid uptake, tissue bioconcentration factor (BCF) and lipid BCF of triclosan and triclocarban were investigated in three developmental stages of X. laevis, and in one developmental stage of B. woodhousii woodhousii, and R. sphenocephala. For most tissue and lipid uptake values, a significant increase was observed as exposure concentration increased. Tissue and lipid BCF values were dependent upon both stage and species. Chronic and acute effects of caffeine were determined in X. laevis larvae. Acute 96-hour LC50 values in four developmental stages were > 75,000 ug L-1 caffeine and heart rates were significantly different at the two earliest developmental stages. Larvae chronically exposed to caffeine reached metamorphosis at the same time as controls. Changes in ...
Date: August 2009
Creator: Palenske, Nicole Marie
Partner: UNT Libraries

Effects of Whisker-Trimming on GABAA Receptors in S1 Cortex

Description: A number of studies have shown that sensory deprivation is associated with selective decreases in GABA, GAD, and GABA receptors, in deprived areas of visual and somatosensory cortex. Those studies focused on layer 4, a recipient of direct thalamocortical sensory input. However, supragranular layers 2/3 have been recently identified as a major locus of functional plasticity in sensory deprivation and long-term potentiation. To examine whether GABAA receptors in layers 2/3 are affected by sensory deprivation, rats had mystacial vibrissae in middle row C or rows ABDE trimmed for 6 weeks beginning in early adulthood. Layers 2/3 above the deprived and adjacent whisker barrels were located in tangential sections, using patterns of radial blood vessels as fiducial marks. In deprived whisker barrel columns, [3H]muscimol binding to GABAA receptors decreased by 12.8% ± 1.2 (P &lt; 0.001) in layers 2/3 and 11.4% ± 1.2 (P<0.001) in layer 4. Altered levels of GABAA α1 subunit (Fritschy et al., 1994) were indicated by reduced optical density of immunostaining, both in deprived layers 2/3 (6.4% ± 0.7; P&lt; 0.001) and in layer 4 (3.4% ± 1.0; P &lt; 0.005). Interestingly, Nissl staining density also decreased in deprived layers 2/3 (12.7% ± 1.8 P &lt; 0.001) and in 4 (6.0 ± 0.7 (P &lt; 0.001). The percent decreases were greater in layers 2/3 than in 4 for both GABAA α1 (P &lt; 0.05) and Nissl substance (P &lt; 0.005). The present results suggest that down-regulation in GABAA receptors may underlie the physiological signs of disinhibition observed in neurons of layer 2/3 and 4 in deprived whisker barrel columns.
Date: August 2004
Creator: Salazar, Eduardo
Partner: UNT Libraries

Endocrine Disruption of Levonorgestrel in Early-life Stages of Fathead Minnows, Pimephales Promelas

Description: Pharmaceuticals have routinely been detected in the environment resulting in a growing concern about whether these drugs could elicit effects on aquatic organisms. The concerns are centered on the highly conserved nature of mammalian therapeutic targets in fish. These pharmaceuticals are found at very low levels in the environment, which can result in sub-lethal effects in aquatic organisms. Therefore, 28 d early-life stage studies were conducted on six pharmaceuticals to assess their impacts on survival and growth fathead minnow larvae. Two pharmaceuticals tested, carbamazepine and fenofibrate, resulted in no alterations to survival and growth. However, amiodarone, clozapine, dexamethasone, and levonorgestrel (LNG) reduced survival at concentrations tested with LNG being the most potent at 462 ng/L. Survival was increased with amiodarone and clozapine; however LNG significantly decreased growth at 86 ng/L. Therefore, the most potent pharmaceutical tested was the synthetic progestin LNG with survival and growth impacts at concentrations less than 1 μg/L. Further analysis was conducted by measuring specific endocrine related mRNA transcript profiles in FHM larvae following the 28 d ELS exposure to LNG. Transcripts of 3β-HSD, 20β-HSD, and FSH were significantly down-regulated following 28 d exposure to both 16.3 and 86.9 ng/L LNG. Also, CYP19a expression was significantly down-regulated at 86.9 and 2392 ng/L LNG. Subsequently, a second study examined time periods that may be most sensitive (e.g., windows of sensitivity) for FHM larvae exposed to LNG. Larvae were exposed to a single concentration of LNG (i.e. LOECgrowth of 86.2 ng/L as determined in the 28 d ELS study) for different time periods starting with fertilized egg through 28 dph. Growth and mRNA expression of the four differentially expressed transcripts from the first study were measured. Regardless of the duration of exposure, LNG significantly decreased growth in fathead minnow larvae at day 28. For both 20β-HSD and CYP19a, ...
Date: August 2013
Creator: Overturf, Matthew D.
Partner: UNT Libraries

Environmental Modulation of the Onset of Air-breathing of the Siamese Fighting Fish and the Blue Gourami

Description: This study determined the effect of hypoxia on air-breathing onset and physiological and morphological characters in larvae of the air breathing fishes Trichopodus trichopterus and Betta splendens. Larvae were exposed intermittently (12/12 h daily) to 20, 17, and 14 kPa of PO2 from 1 to 40 days post-fertilization. Survival, onset of air breathing, wet body mass, O2, Pcrit were measured every 5 dpf. Hypoxia advanced by 4 days, and delayed by 9 days, the onset of air breathing in Betta and Trichopodus, respectively. Hypoxia increased larval body length, wet mass, and labyrinth organ respiratory surface of Betta, but did not affect these factors in Trichopodus. Hypoxic exposure increased O2 by 50-100% at each day throughout larval development in Betta, but had no effect on larval Trichopodus. Hypoxia decreased Pcrit in Betta by 37%, but increased Pcrit in Trichopodus by 70%. Larval Betta reared in hypoxia showed a modified heart rate:opercular rate ratio (3:1 to 2:1), but these changes did not occur in Trichopodus. Compared to Betta, the blood of Trichopodus had a higher P50 and much smaller Bohr and Root effects. These interspecific differences are likely due to ecophysiological differences: Betta is a non- obligatory air-breather after 36 dpf with a slow lifestyle reflected in its low metabolism, while Trichopodus is an obligatory air-breather past 32 dpf with an athletic fast lifestyle and accompanying high metabolism.
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Date: December 2015
Creator: Mendez Sanchez, Jose Fernando
Partner: UNT Libraries

Enzyme Assays Using Earthworms for Assessing Innate and Nonspecific Immunotoxicity of Xenobiotics

Description: Principal objectives of my research were to: (1) report for the first time that coelomocytes are able to reduce NBT dye and confirm the presence of lysozyme-like activity in earthworm; (2) develop a standard methodology for determination of NBT reduction and lysozyme-like activity in earthworms; (3) compare NBT reduction and lysozyme-like activity in earthworms with those of murine and human cells and fluids; and (4) demonstrate the sensitivity of earthworm NBT reduction and lysozyme-like activity as the assays using matrics in refuse-derived fuel fly ash (RDFF) and CuSO4.
Date: May 1992
Creator: Chen, Shing-Chong
Partner: UNT Libraries

The Eosinophil Response in Mice Infected with Trichinella spiralis or Trichinella pseudospiralis as Indicated by Phospholipase B Activity

Description: The host eosinophil response was compared in mice infected with either T. spiralis or T. pseudospiralis by determination of levels of splenic and intestinal phospholipase B, a marker enzyme for eosinophils. Primary infection of naive mice and challenge infection of homologously sensitized mice with T. pseudospiralis resulted in significantly lower tissue phospholipase B activities than infection with T. spiralis. Mice homologously challenged with T. pseudospiralis did exhibit an anamnestic eosinophil response compared to mice given a primary T. pseudospiralis infection. This anamnestic response, however, was significantly lower than the eosinophil response seen in sensitized mice given a homologous T. spiralis challenge. Mice sensitized to T. spiralis or T. pseudospiralis and heterologous challenge demonstrated an elevated eosinophil response compared to mice given a primary infection with either parasite. The heterologous challenge response, however, was not as intense as found for sensitized mice given a homologous challenge.
Date: December 1992
Creator: Hsu, Shing-Chien
Partner: UNT Libraries

Establishing genetic and physiological baselines for the black-tailed prairie dog (Cynomys ludovicianus).

Description: The black-tailed prairie dog (Cynomys ludovicianus) has experienced dramatic declines over much of its historical range due to habitat loss, plague, poisonings, and shootings. Many populations now occur as isolated genetic relicts. A multiple locus genetic profile was obtained using microsatellite analyses of six polymorphic nucleotide repeats from 319 black-tailed prairie dogs collected from 16 colonies throughout the state of Texas. This assessment revealed that existing populations have sufficient variation at all six loci to verify the usefulness of this approach as a primary genetic tool in conservation and preservation. The data reveals regional-dependent frequency patterns as well as support for founder/bottleneck effects for several of the 16 sites. Although the prairie dog population in Texas as a whole may appear genetically diverse, considerable genetic divergence has already occurred among the subpopulations (FST = 0.164). Isolation by distance is supported by genic differentiation analysis (P < 0.001) and pairwise correlation analysis between genetic distance and geographic distance (P < 0.001). Prairie dogs from six (COC, LUBA, LUBC, LUBD, LUBE, and TAR) of the original 16 sites have been relocated or exterminated, or were in the process of being relocated. Results indicated the following colonies (COT, DAL, HOW, and HUD) are of sufficient size and possess ample genetic diversity to be characterized as candidate foundation populations for future preservation efforts. The proximity of small colonies (< 20 hectares) such as HEMB, LUBB, and PEC, to other colonies should be examined to determine if they are isolated or part of a metapopulation. Colonies (HAR, HEMA, and SCH) with low genetic diversity would be ideal candidates for supplementation. Alternatively, these colonies could be relocated or blended with other similar but genetically distinct colonies. Baselines for healthy, pet prairie dog hematology and blood chemistries were also established. Results signify that data gathered from pet ...
Date: May 2007
Creator: Biggs, Cindy Dawn
Partner: UNT Libraries

Eutrophication Monitoring and Prediction

Description: Changes in trophic status are often related to increases or decreases in the allocthonous inputs of nutrients from changes in land use and management practices. Lake and reservoir managers are continually faced with the questions of what to monitor, how to monitor it, and how much change is necessary to be considered significant. This study is a compilation of four manuscripts, addressing one of these questions, using data from six reservoirs in Texas.
Date: December 1993
Creator: Cairns, Stefan H., 1949-
Partner: UNT Libraries

Evaluation of City of Denton Sub-Watershed by Benthic Macroinvertebrate Field Experimental Approach

Description: In this study, two different field experiments were designed to assess the relative influence of urbanization on benthic communities. During spring and summer, four urban and one reference sites from Denton County, Texas were selected for benthic macroinvertebrate evaluation. Statistically significant differences in colonized benthic macroinvertebrate taxa on artificial substrates were observed among the four urban sites and the reference site. Oligochaetes and chironomids were the dominant taxa at all sites. Identification of chironomid larvae at the subfamily and genus level to detect differences between sites had higher statistical power than the evaluation based on total chironomids. At the reference site, Caenis, Cladotanytarsus, Orthocladius, and Ceratopogonidae were the dominant taxa, while the urban sites were dominated by Dero, Physella, Ancylidae, Chironomus, Dicrotendipes, Glyptotendipes, Polypedilum, Pseudochironomus, Stenochironomus, and Tanytarsus. These differences may have been dependent upon differences in hydrologic regime and water quality between sites. Significant differences (ANOVA, p < 0.01) in water quality parameters (alkalinity, hardness, nitrates, phosphates, chlorides, sulfates, calcium, magnesium, potassium, and triazine) were found among water samples collected from the reference and urban sites. During the transfer period, most of the Ephemeroptera and Trichoptera taxa and a few other taxa disappeared from artificial substrates that were colonized at the reference site and then moved to the urban sites. Also, local abundant taxa from the urban site significantly (t test, p < 0.05) increased in number on the transferred artificial substrates. Seasonal differences in colonization patterns were also observed between the spring and summer experimental periods, which indicate that temporal variation is equally important, as is the anthropogenic effect in benthic community evaluation. Field survival and growth experiments using Erpetogomphus designatus larvae were designed to detect differences between evaluated sites. Larvae were collected from the reference site, measured in the laboratory, and exposed at the urban sites for ...
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Date: August 2006
Creator: Mahato, Mahendra
Partner: UNT Libraries

Evolutionary Genetics of Certain Mice of the Peromyscus boylii Species Group

Description: The genetic structure of 49 natural populations of four species (P. attwateri, P. boylii, P. pectoralis, and P. polius) of the Peromyscus boylii species group was analyzed through application of chromosomal and electrophoretic techniques. Chromosomal variation within and among populations of the boylii species group was analyzed from 178 specimens. Electrophoretic techniques were utilized for the demonstration of variation in enzymes and other proteins encoded by structural loci and applied to the study of the evolution of the boylii species group by estimation of levels of genetic heterozygosity within populations, estimation of degree of genetic similarity between conspecific populations and between species, and determination of patterns of geographic variation in allelic frequencies and levels of heterozygosity. Six distinct chromosomal patterns were observed among the populations of the four species of the boylii species group. All specimens had a diploid number of 48 and the major difference in chromosomal morphology was in the number of pairs of large to medium biarmed autosomes. Little or no chromosomal variation was observed in three species (attwateri, pectoralis and polius), but considerable chromosomal variation occurred among populations of P. boylii. Generally, the chromosomal variation in P. bylii was between allopatric populations, with each chromosomal pattern limited to a recognized subspecies. Polymorphism was observed in two populations. The polymorphism observed in P. polius was the result of pericentric inversion involving the smallest pair of metacentric autosomes. The polymorphism observed in P. bolii cileus was interpreted at the result of gene flow between P. boylii rowleyi and P. boylii spicilegus. In addition to chromosomal evidence, analysis of electrophoretic data demonstrated and suggested effective gene flow between the chromosomal forms of P. boylii. Electrophoretically demonstrable variation was analyzed in 11 proteins encoded by 17 autosomal loci. Of the 17 structural loci, 11 were polymorphic in one or more ...
Date: December 1973
Creator: Kilpatrick, Charles William
Partner: UNT Libraries

Field and Laboratory Fish Tissue Accumulation of Carbamazepine and Amiodarone

Description: The goals of this dissertation work were to assess the bioaccumulation potential of carbamazepine and amiodarone, two widely used ionizable pharmaceutical compounds that possess mid-range and high LogD values, respectively, and to evaluate alternative methods to assess chemical accumulation in bluntnose minnows, catfish, and tilapia. Results indicated that carbamazepine does not appreciably bioaccumulate in fish tissue with BCFk and BAF carbamazepine values < 10. Amiodarone, however, with a log D of 5.87 at pH 7.4, accumulated in fish tissues with kinetic BCF values <2,400. Collectively, the data suggest that full and abbreviated laboratory-derived BCFs, BCFMs derived from S9 loss-of-parent assays, as well as field BAF values are similar for each of the two drugs. In summary, the results from this dissertation indicated: 1) The reduced design BCF test is a good estimate for the traditional OECD 305 test. 2) In vitro S9 metabolism assays provide comparable BCF estimates to the OECD 305 test. 3) Metabolism may play a large role in the accumulation of drugs in fish. 4) Reduced BCF tests and in vitro assays are cost effective and can reduce vertebrate testing.
Date: December 2013
Creator: García Martínez, Santos Noé
Partner: UNT Libraries

Fitness-Related Alterations in Blood Pressure Control: The Role of the Autonomic Nervous System

Description: Baroreflex function and cardiovascular responses to lower body negative pressure during selective autonomic blockade were evaluated in endurance exercise trained (ET) and untrained (UT) men. Baroreflex function was evaluated using a progressive intravenous infusion of phenylephrine HCL (PE) to a maximum of 0.12 mg/min. Heart rate, arterial blood pressure, cardiac output and forearm blood flow were measured at each infusion rate of PE. The reduction in forearm blood flow and concomitant rise in forearm vascular resistance was the same for each subject group. However, the heart rate decreases per unit increase of systolic or mean blood pressure were significantly (P<.05) less in the ET subjects (0.91 ± 0.30 versus 1.62 ± 0.28 for UT). During progressive lower body negative pressure with no drug intervention, the ET subjects had a significantly (P<.05) greater fall in systolic blood pressure (33.8 ± 4.8 torr versus 16.7 ± 3.9 torr). However, the change in forearm blood flow or resistance was not significantly different between groups. Blockade of parasympathetic receptors with atropine (0.04 mg/kg) eliminated the differences in response to lower body negative pressure. Blockade of cardiac sympathetic receptors with metoprolol (0.02 mg/kg) did not affect the differences observed during the control test. It was concluded that the ET subjects were less effective in regulating blood pressure than the UT subjects, because of 1) an attenuated baroreflex sensitivity, and 2) parasympathetic-mediated depression of cardiac and vasoconstrictive responses to the hypotensive stress.
Date: December 1986
Creator: Smith, Michael Lamar, 1957-
Partner: UNT Libraries

Food, Feeding Selectivity, and Ecological Efficiencies of Fundulus notatus (Rafinesque) (Osteichthyes; Cyprinodontidae)

Description: This study was made to further define the trophic dynamics of Fundulus notatus by determining its ration composition under natural conditions, measuring feeding selectivity under various laboratory conditions of prey-species composition and availability, and determining the efficiencies with which F. notatus utilizes ingested chironomid larvae.
Date: August 1970
Creator: Atmar, Gerald Legare
Partner: UNT Libraries

Genetic Mechanisms for Anoxia Survival in C. Elegans

Description: Oxygen deprivation can be pathological for many organisms, including humans. Consequently, there are several biologically and economically relevant negative impacts associated with oxygen deprivation. Developing an understanding of which genes can influence survival of oxygen deprivation will enable the formulation of more effective policies and practices. In this dissertation, genes that influence adult anoxia survival in the model metazoan system, C. elegans, are identified and characterized. Insulin-like signaling, gonad function and gender have been shown to influence longevity and stress resistance in the soil nematode, C. elegans. Thus, either of these two processes or gender may influence anoxia survival. The hypothesis that insulin-like signaling alters anoxia survival in C. elegans is tested in Aim I. The hypotheses that gonad function or gender modulates anoxia survival are tested in Aim II. Insulin-like signaling affects anoxia survival in C. elegans. Reduction of insulin-like signaling through mutation of the insulin-like receptor, DAF-2, increases anoxia survival rates in a gpd-2/3 dependent manner. The glycolytic genes gpd-2/3 are necessary for wild-type response to anoxia, and sufficient for increasing anoxia survival through overexpression. Gonad function and gender both affect anoxia survival in C. elegans. A reduction of ovulation and oocyte maturation, as measured by oocyte flux, is associated with enhanced anoxia survival in all cases examined to date. Reduction of function of several genes involved in germline development and RTK/Ras/MAPK signaling reduce ovulation and oocyte maturation while concurrently increasing anoxia survival. The act of mating does not influence anoxia survival, but altering ovulation through breeding or chemical treatment does. The male phenotype also increases anoxia survival rates independent of genotype. These studies have identified and characterized over ten different genotypes that affect adult survival of anoxia in C. elegans. Before these studies were conducted, there were no genes known to influence adult anoxia survival in C. ...
Date: August 2008
Creator: Mendenhall, Alexander R.
Partner: UNT Libraries

Glucose and Altered Ceramide Biosynthesis Impact the Transcriptome and the Lipidome of Caenorhabditis elegans

Description: The worldwide rise of diabetes and obesity has spurred research investigating the molecular mechanisms that mediate the deleterious effects associated with these diseases. Individuals with diabetes and/or obesity are at increased risk from a variety of health consequences, including heart attack, stroke and peripheral vascular disease; all of these complications have oxygen deprivation as the central component of their pathology. The nematode Caenorhabditis elegans has been established as a model system for understanding the genetic and molecular regulation of oxygen deprivation response, and in recent years methods have been developed to study the effects of excess glucose and altered lipid homeostasis. Using C. elegans, I investigated transcriptomic profiles of wild-type and hyl-2(tm2031) ( a ceramide biosynthesis mutant) animals fed a standard or a glucose supplemented diet. I then completed a pilot RNAi screen of differentially regulated genes and found that genes involved in the endobiotic detoxification pathway (ugt-63 and cyp-25A1) modulate anoxia response. I then used a lipidomic approach to determine whether glucose feeding or mutations in the ceramide biosynthesis pathway or the insulin-like signaling pathway impact lipid profiles. I found that gluocose alters the lipid profile of daf-2(e1370) (an insulin-like receptor mutant) animals. These studies indicate that a transcriptomic approach can be used to discover novel pathways involved in oxygen deprivation response and further validate C. elegans as a model for understanding diabetes and obesity.
Date: August 2016
Creator: Ladage, Mary Lee
Partner: UNT Libraries

Glucose Induces Sensitivity to Oxygen Deprivation and Alters Gene Expression in Caenorhabditis Elegans

Description: An organisms’ diet represents an exogenous influence that often yields colossal effects on long-term health and disease risk. The overconsumption of dietary sugars for example, has contributed to significant increases in obesity and type-2 diabetes; health issues that are costly both economically and in terms of human life. Individuals who are obese or are type-2 diabetic often have compromised oxygen delivery and an increased vulnerability to oxygen-deprivation related complications, such as ischemic strokes, peripheral arterial disease and myocardial infarction. Thus, it is of interest to identify the molecular changes glucose supplementation or hyperglycemia can induce, which ultimately compromise oxygen deprivation responses. By utilizing the Caenorhabditis elegans genetic model system, which is anoxia tolerant, I determined that a glucose-supplemented diet negatively impacts responses to anoxia and that the insulin-like signaling pathway, through fatty acid and ceramide biosynthesis and antioxidant activity, modulates anoxia survival. Additionally, a glucose-supplemented diet induces lipid accumulation. Use of RNA-sequencing analysis to compare gene expression responses in animals fed either a standard or glucose-supplemented diet revealed that glucose impacts the expression of genes involved with multiple cellular processes including lipid and carbohydrate metabolism, stress responses, cell division, and extracellular functions. Several of the genes we identified are homologous to human genes that are differentially regulated in response to metabolic diseases, suggesting that there may be conserved gene expression responses between C. elegans supplemented with glucose and a diabetic and/or obese state observed in humans. These findings support the utility of C. elegans to model specific aspects of the T2D disease process (e.g., glucose-induced sensitivity to oxygen deprivation) and identify potentially novel regulators of common complications seen in hyperglycemic and T2D patients (e.g., macrovascular complications).
Date: August 2015
Creator: Garcia, Anastacia M.
Partner: UNT Libraries

Head Trauma Release of Histamine from Dural Mast Cells Alters Blood-Brain Barrier: Attenuation with Zolantidine

Description: This study employed a new model of mild-to-moderate head trauma to specifically identify the role of dural mast cell (MC) histamine in trauma-induced increased permeability in the blood-brain barrier (BBB). A single line was scored partially through the left dorsal parietal skull. Immediately following the trauma, degranulation was seen in 39% of the MCs on the left and in 2% on the right. After a 20 min survival period, left duras showed 55% with MC degranulation (fewer with complete degranulation) compared to 34% on the right. In the other experiments two parallel lines were scored following the injection of Evan's blue. Histamine assay showed histamine increased in the left cortex to 154% at 5 min, 174% at 10 min, and 151% at 20 min. Fluorescent quantitation of extravasated Evan's blue at 20 min following the trauma gave an increase of 1385% over the value measured for the right cortex. Zolantidine, a selective histamine H2 receptor antagonist, administered at 10- and 20- mg/kg 30 min before the trauma blocked 65% of the Evan's blue extravasation compared with the control and 2.5 mg group.
Date: December 2000
Creator: Laufer, Susan R.
Partner: UNT Libraries

Hypoxic and hyperoxic incubation affects the ductus arteriosus in the developing chicken embryo (Gallus gallus).

Description: Developing chicken embryos have two ductus arteriosus (DA) that shunt blood away from the lungs and to the chorioallantoic membrane, the embryonic gas exchanger. In mammals, DA closure is stimulated by an increase in blood gas O2 that occurs as the animal begins to breathe with its lungs. The goal of this study was to determine the influence of O2 levels during incubation on the vascular reactivity and morphology of the O2-sensitive DA and to examine the effects of changing O2 levels during late incubation on the morphology of the DA from chicken embryos. In comparison to normoxia, hypoxia (15%) reduced venous O2 levels in day 16 and day 18 embryos and reduced aircell O2 values in day 16, day 18, and internally pipped (IP) embryos, whereas hyperoxia (30%) increased venous O2 levels and aircell O2 level in day 16, day 18, and IP embryos. In comparison to normoxia, hypoxia delayed closure of the DA, whereas hyperoxia accelerated DA closure. In comparison to the left DA from externally pipped (EP) normoxic embryos, the left DA from EP hypoxic embryos exhibited a significantly weaker contractile response to O2. The DA from day 18 hypoxic embryos exhibited a significantly weaker contractile response to norepinephrine and phenylephrine when compared with the DA from day 18 normoxic and hyperoxic embryos. The effect of incubation in hypoxia / hyperoxia during different developmental windows on the DA O2-induced contractile response was observed only in IP embryos that were incubated in normoxia for 16 days and were then moved to hyperoxia. Incubation in hypoxia / hyperoxia resulted in differences in embryo mass, yolk mass, and heart mass. There is an association between the decreased contractile response to O2 and delayed closure in the proximal portion of the DA from hypoxic embryos; as well as an increased contractile ...
Date: December 2009
Creator: Copeland, Jennifer
Partner: UNT Libraries