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- Nyrén, Rakel, 1986-
(author)
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Localization of lipoprotein lipase in mouse pancreas, kidney and placenta : impact of metabolic disturbances on cellular distribution and activity regulation
- 2020
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Doctoral thesis (other academic/artistic)abstract
- Lipoprotein lipase (LPL) is the key enzyme for metabolism of triglycerides in plasma lipoproteins. In recent years many new facts about the enzyme and its regulation have been uncovered. The endothelial membrane protein GPIHBP1 translocates LPL through endothelial cells and holds the enzyme in place at the luminal side of the capillary endothelium. Some of the angiopoietin-like proteins (ANGPTLs) bind to LPL and are responsible for tissue-specific regulation of the enzyme’s catalytic activity. Most studies in the past have focused on LPL in adipose and muscle tissues. LPL is also present in several other tissues, but the localization and function of LPL at these sites have not been fully elucidated.One aim of the present thesis was to develop a protocol for immunolocalization of LPL in mouse tissues. In pancreas, the enzyme was localized to capillaries of the exocrine tissue, together with GPIHBP1, but also inside α- and β-cells. LPL in β-cells was absent in leptin-deficient ob/ob mice, but appeared after treatment with leptin. In kidney, LPL was mostly present within the proximal tubular cells of the nephron. In fed animals, LPL was also seen in intertubular vessels together with GPIHBP1. A LPL knock-out mouse model, MCKL0, was used to validate the specificity of our immuno-protocol. Kidneys from these mice showed no or very little staining for LPL. In mouse placenta, LPL was mostly found in capillaries of the labyrinth zone, where the exchange between fetal and maternal blood occurs.A second aim was to gain better understanding for when, how and why LPL activity is regulated in mouse kidneys, and how obesity induced by high-fat diet (HFD) affects the LPL system. LPL activity in kidneys was regulated by ANGPTL4 in a similar manner as LPL in white adipose tissue, but in contrast to adipose tissue, the kidney LPL did not contribute to the uptake of fatty acids from chylomicron triglycerides. We found that obesity and insulin resistance, induced by long-term feeding of HFD, abolished the nutritional regulation of LPL activity in kidneys of male, but not of female, mice. To directly study the uptake of energy substrates in mouse kidneys, we developed a protocol for measurement of radiolabeled substrates in kidneys using PET/CT with the tracers [18F]FDG (a glucose analogue) and [18F]FTHA (a fatty acid analogue) injected to blood. There was an increase in uptake of both tracers in fasted male mice that had been on long-term HFD, compared to controls, as revealed by scanning of perfused organs, ex vivo, 3 hours after the injections.A third aim was to study LPL and the function of ANGPTL4 in pregnant mice and placentas. ANGPTL4 is known to increase in human plasma throughout pregnancy. As ANGPTL4 levels rise, triglyceride levels increase as well. We used mice that either lacked (Angptl4-/-) or overexpressed Angptl4 (Angptl4-tg+/-), and compared them to wild-type mice. Plasma triglycerides and VLDL levels increased during pregnancy both in wild-type and in Angptl4-/- mice. The lipid profile in Angptl4-tg+/- was high already before conception, and did not change. LPL activity in placenta was, however, similar in all genotypes. The increase in ANGPTL4 in maternal blood during pregnancy might originate from placenta, but Angptl4 expression was also increased in maternal liver and subcutaneous white adipose tissue. The pups from Angptl4-tg+/- had reduced birthweight compared to pups from wild-type and Angptl4-/- mice.In conclusion, the present thesis provides information on the localization and possible functions of LPL and some of its regulator proteins in mouse pancreas, kidney and placenta. New data on the regulation of LPL activity in mouse kidney, and the effects of HFD and obesity, is presented, as well as insights into the potential role of ANGPTL4 for control of plasma triglyceride levels and fetal growth during mouse pregnancy.
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