Browsing by Subject "Excretion"
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Item Cloning, Immunolocalization and Functional Analyses of Calcitonin Receptor 1 (AedaeGPRCAL1; Diuretic Hormone 31 Receptor) in Females of Mosquito Aedes aegypti (Diptera: Culicidae)(2013-04-26) Kwon, Hyeog SunG protein-coupled receptors (GPCRs) are composed of seven transmembrane domains and play an essential role in regulating physiological functions and mediating responses to environmental stimuli, biogenic amines, neurotransmitters, peptides, lipids, and hormones. The calcitonin-like diuretic hormone 31 (DH31) is known to elicit natriuresis from the Malpighian tubules (MTs) of mosquitoes Anopheles gambiae and Aedes aegypti upon blood feeding. However, the contribution of DH31 cognate receptor, calcitonin receptor 1 (GPRCAL1), has not been evaluated with respect to postprandial fluid regulation or myostimulatory activity in blood feeding insects. Thus, this dissertation has investigated potential roles of AedaeGPRCAL1 in the regulation of fluid homeostasis and hindgut muscle contraction in female A. aegypti mosquito. The full length cDNA encoding AedaeGPRCAL1 was cloned and sequenced. The receptor expression in the MTs and hindgut from female mosquito was analyzed by western blot and immunohistochemistry using anti-AedaeGPRCAL1 affinity purified antibodies, and subsequently its role in fluid transport and hindgut contraction was evaluated by RNA interference (RNAi). The mosquitoes that underwent knock-down of the AedaeGPRcal1 exhibited up to 57% lower rate of MT fluid secretion in presence of Aedae-DH31 in the in vitro assay and a ~30% reduction in the fluid excreted from live females upon blood feeding. The receptor was immunolocalized in principal cells, predominantly towards the distal end of MTs. Analyses of receptor signal probability indicate the receptor is expressed in a gradient-like fashion along the length of the MTs. A striking discovery was the fact that not all principal cells express the receptor, contrary to previous belief. Immunolocalization revealed the AedaeGPRCAL1 is expressed in hindgut circular and longitudinal muscles. The application of DH31 increased the frequency of hindgut contractions in all female mosquitoes, those injected with AedaeGPRcal1 dsRNA and controls, as compared to their basal contraction rate, but the percent change in frequency of hindgut contraction from AedaeGPRcal1 knock-down females was about 2-fold lower than the controls after application of Aedae-DH31. To my knowledge, this is first evidence of RNAi-induced phenotypes in any invertebrate that allowed the quantification of the contribution of single family B GPCR to fluid loss and muscle contractility.Item Increased temperature effects on fish-mediated nutrient cycling in an East Texas stream(2017-11-07) McWilliams, Jessica Lynn; Hargrave, Chad W.The unprecedented rate of global warming is an inevitable outcome of anthropogenic CO2 release into the atmosphere and complex climatic feedbacks. In ectotherms, increasing temperature may increase metabolic rates, which could enhance the energy demands of individuals and should accelerate resource acquisition. Population size and fish biomass were measured seasonally in a small second order stream over a 10-year period to examine seasonal variation in these parameters. I examined effects of increased temperature on nitrogen and phosphorus excretion in the four most abundant fish in this stream system. These fishes represent three functional feeding guilds common to many temperate stream ecosystems and comprise approximately 80-90% of the fish community. I developed temperature dependent nitrogen and phosphorus excretion models for fishes and applied these models to daily average temperatures in the stream. I then simulated climate warming (+2, +4, & +6°C) to examine the potential effects of increased temperature on fish-mediated nutrient dynamics in a southern temperate stream ecosystem. I found that increased temperature does increase nutrient cycling and nutrient flux within aquatic ecosystems; however, these effects appear to be tied to population size, biomass in addition to seasonal temperature. With increased temperature effects in spring and autumn having the greatest effect, when temperatures are cool and fish abundance and biomass is also greatest.