Browsing by Subject "aineenvaihdunta"
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(2019)Glutamine, the conditionally essential amino acid, is a major carbon and nitrogen carrier required for a range of cell functions, such as protein synthesis and maintaining redox balance. While healthy cells adjust their activities in response to glutamine availability, tumor cells display deregulated glutamine uptake and metabolism allowing quick proliferation and survival in cellular stress conditions. Hence, further knowledge of the glutamine sensing network is of interest. Utilizing Drosophila melanogaster, the roles of formerly identified glutamine sensing regulator candidates, Forkhead box O (FoxO), Super sex combs (Sxc), Spalt major (Salm) and Spalt-related (Salr), were explored. Drosophila is an efficient model organism for analyzing gene regulatory mechanisms, with its simple genome but conserved genes and metabolic pathways. Loss-of function and gain-of-function mutants of the candidates were cultured with/without glutamine, and their physiological response and gene expression changes were analyzed. The results show the glutamine intolerant phenotype of FoxO and Sxc deficiency, not dependent on altered food intake levels of larvae. However, glutamine intolerance of Salr and Salm deficiency was not observed. Moreover, we aimed to gain further insight to the roles of FoxO and Sxc in glutamine metabolism. Since amino acid catabolism produces ammonia, and glutamine metabolism plays a vital role in ammonia detoxification, we performed a pH-based measurement of foxo and sxc mutant larvae hemolymph on food with/without glutamine. However, we could not associate FoxO or Sxc with regulation of glutamine-derived ammonia clearance. In addition, we explored FoxO downstream regulator candidates. Putative promoter areas of Paics, Uro, Sesn, salr, Prat2 and Gdh were cloned into reporter vectors and the luciferase activity was analyzed under the expression of foxo. The results indicate that FoxO is a regulator of all of the 6 genes. Next we could utilize the here constructed plasmids to see whether the FoxO-mediated regulation is affected by altered glutamine levels in cell culture.
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(2022)Mercury is a toxic heavy metal that ends up in aquatic ecosystems both as atmospheric deposition as well as from the catchment area surrounding the water body. Under the right circumstances, inorganic mercury can be methylated into methylmercury, which accumulates in organisms and food webs and is harmful to humans and animals. Humans are exposed to methylmercury mainly through consumed fish as almost all the total mercury content in fish muscle consist of methylmercury. Mercury content of fish is affected by numerous different factors, such as the food consumed by the fish, the characteristics of the fish species and fish individual, level of lake eutrophication, mercury concentration in water and catchment area. Thesis studies total mercury content and mercury bioaccumulation in cyprinid fish. The species included in the study are roach (Rutilus rutilus), bream (Abramis brama), bleak (Alburnus alburnus), white bream (Blicca bjoerkna), blue bream (Ballerus ballerus), ide (Leuciscus idus), tench (Tinca tinca), rudd (Scardinius erythrophthalmus) and crucian carp (Carassius carassius). Three main research questions were: (1) Can diet of different cyprinid fish explain the mercury content? (2) Is mercury content consistent between fish species in different lake types, or does lake type affect mercury content? (3) Is mercury bioaccumulation consistently similar between fish species between lake types? The fish samples for this study were gathered prior to MSc during July-August in 2020 and 2021 from Kukkia, Hauhonselkä and Pääjärvi, which are classified as mesotrophic lakes, and from eutrophic Vesijärvi, Tuusulanjärvi and Hulausjärvi. Total length, weight, sex, sexual maturity, condition factor (K), fish diet and muscle total mercury content were determined from each fish. The diet of the fish and the differences in the total mercury content were tested with analysis of variance. Mercury bioaccumulation was tested with using linear regression analysis and the slope coefficient obtained from the regression equation was tested for bleak and roach with t-test. From the abundant species of the study the highest length corrected (to length 16.6 cm) mercury contents were measured in bleak and roach, while the contents were lower in white bream and bream. The mercury content differed most clearly in species whose feeding behaviour was very specialized. For example, the differences in mercury contents between bleak, which mainly consumes surface insects and zooplankton, and bream, which prefers benthic food, were clear in every lake studied. Mercury contents were mainly higher in mesotrophic lakes compared to eutrophic lakes, but results varied by species. In addition to fish diet and lake type, fish’s metabolism, growth rate and life cycle length probably influenced the species-specific mercury contents. When comparing the abundant species, the bioaccumulation of mercury was steepest in bleak and roach. Compared to these species’ bioaccumulation was slower in white bream and bream. When comparing mercury bioaccumulation within species between different lake types, the differences were species-specific. For example, the bioaccumulation of bleak was consistently steeper in mesotrophic lakes compared to eutrophic lakes, but the bioaccumulation of white bream was steeper in eutrophic lakes. Based on this study, it can be concluded that mercury content and bioaccumulation in cyprinid fish were influenced by e.g., species-specific characteristics, fish diet and level of the lake productivity. In addition to these factors, mercury content and bioaccumulation of fish may have been affected by the land use of the lake catchment area, as well as the anoxic areas of the lakes, which are propitious places for methylation. This study provided valid information on which factors may be relevant for cyprinid fish when observing their mercury content and bioaccumulation.
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