20.500.12556/RUL-169076 Vpliv presnove lipidnih kapljic na morfologijo in količino mitohondrijev v stradanih celicah Impact of lipid droplet metabolism on mitochondrial morphology and abundance in starved cells Mitohondriji so odgovorni za vzdrževanje energijske homeostaze preko razgradnje hranil, hkrati pa tudi podpirajo rast celic preko proizvodnje gradnikov potrebnih za biosintezo proteinov, lipidov in nukleinskih kislin. Nedavne raziskave so pokazale, da spremembe v funkciji mitohondrijev odsevajo razlike v njihovi dinamiki. Ta termin se nanaša na spremembe v količini, številu, morfologiji in relativnem položaju mitohondrijev znotraj celice. Interakcije med mitohondriji in lipidnimi kapljicami so nujno potrebne za oksidacijo maščobnih kislin in proizvodnjo energije, hkrati pa tudi podpirajo sintezo lipidov in tvorbo lipidnih kapljic. Kljub temu mehanizmi, ki povezujejo funkcije mitohondrijev in lipidnih kapljic znotraj različnih celičnih okolij ostajajo slabo poznani. Izziv v tej nalogi je bil ugotoviti, če so spremembe v presnovi lipidnih kapljic povezane z jasnim odzivom dinamike mitohondrijev. Za vzpostavitev metod za analizo mitohondrijske dinamike smo z modulacijo dostopnosti hranil izzvali specifične spremembe v njihovi funkciji in nato s pomočjo pretočne citometrije, konfokalne mikroskopije in analize slik kvantificirali vsebnost mitohondrijev, njihovo obliko, velikost in interakcije z lipidnimi kapljicami. Na celični liniji MDA-MB-231 smo uporabili genetske in farmakološke pristope za vpliv na ključna vozlišča v metabolizmu lipidnih kapljic in izmerili spremembe v dinamiki mitohondrijev. Naši rezultati kažejo, da imata glavna mehanizma razgradnje lipidnih kapljic – lipoliza in lipofagija – različne učinke na dinamiko mitohondrijev, kar morda razkriva razlike v funkcijah mehanizmov razgradnje lipidnih kapljic. Mitochondria are responsible for the maintenance of energy homeostasis by breaking down nutrients while also supporting cell growth through the generation of building blocks required for the biosynthesis of proteins, lipids and nucleic acids. Recent experimental work has shown that changes in mitochondrial function are mirrored by alterations in mitochondrial dynamics. This term refers to changes in mitochondrial quantity, number, morphology, and position relative to other cellular structures. Interactions of mitochondria with lipid droplets are required for fatty acid oxidation and energy production, but these interactions also support the synthesis of lipids and their accumulation in lipid droplets. However, the mechanisms that control the coupling of the functions of lipid droplets and mitochondria in various cellular environments remain poorly known. The challenge in this work was to find out whether changes in lipid droplet metabolism are associated with distinct alterations in mitochondrial dynamics. To establish methods for the analysis of mitochondrial dynamics in our lab, we induced specific changes in mitochondrial function by modulating cellular nutrient conditions and used flow cytometry, confocal microscopy and image analysis to quantify mitochondrial content, their shape, size and interactions with lipid droplets. We then used genetic and pharmacological approaches in the MDA-MB-231 cell line to modulate key nodes in lipid droplet metabolism and measured the resulting changes in mitochondrial dynamics. Our results demonstrate that the two primary mechanisms of lipid droplet breakdown – lipolysis and lipophagy – have distinct influences on mitochondrial dynamics, revealing a possible functional separation of lipid droplet breakdown mechanisms. celična kultura lipidne kapljice avtofagija mitohondriji mitofagija zlivanje mitohondrijev kontaktna mesta stradanje cell culture lipid droplets autophagy mitochondria mitophagy mitochondrial fusion contact sites starvation true false false [G. Jazbec] Slovenski jezik Angleški jezik Magistrsko delo/naloga 2025-05-10 07:15:43 2025-05-10 07:15:49 2025-05-12 07:43:27 0000-00-00 00:00:00 2025 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 1970-01-01 576.311.347:616.393(043.2) 257337 235460867 47819.pdf 47819.pdf 1 D5A214E0CF3D8AC0538F7EC275576CE3 f71499c6cfef0b04a61fb48f35bb960547d6044d2af9b570065f06bda6466cf4 a1b64eae-2d5c-11f0-b232-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=210227 Biotehniška fakulteta 0 0 0