Odziv kvasovk vrste Saccharomyces cerevisiae na tvorbo agregatov proteina TDP-43.

Jug, Blaž (author), Jamnik, Polona (mentor), Dogša, Iztok (comentor)

Kvasovke Saccharomyces cerevisiae imajo pomembno vlogo kot modelni organizem za razumevanje razvoja različnih nevrodegenerativnih bolezni. Prav njihova enostavnost za kultivacijo in zadostna podobnost celicam sesalcev sta pripomogla k njihovi splošni uporabi pri različnih znanstvenih študijah. Nevrodegenerativne bolezni predstavljajo težavo in izziv modernega časa, saj so rezultat staranja populacije in podaljševanja življenjske dobe ljudi. Med redke nevrodegenerativne bolezni prištevamo amiotrofično lateralno sklerozo (ALS). Prizadene motorične nevrone v možganih in hrbtenjači ter povzroči njihovo degeneracijo s citoplazemskimi agregati proteina TDP-43. Cilj naloge je bil preučiti odziv kvasovk na prisotnost citotoksičnih agregatov proteina TDP-43-GFP. Kvasovke, ki smo jih gojili v zaporednih šaržnih kultivacijah, smo opazovali s pomočjo fluorescenčne mikroskopije v različnih fazah rasti. Hkrati smo merili fluorescenčno intenziteto za indikacijo izražanja vstavljenega humanega gena TDP-43-GFP, prirast biomase za vpliv agregatov na celično rast in razmnoževanje ter metabolno aktivnost celic kot odraz energijskih potreb za celični stresni odziv na prisotnost agregatov. Z eksperimenti smo razbrali stresni vpliv agregatov na celice in posledično njihov odziv. Ugotovili smo, da je kultura, ki je bila prvotno kultivirana v ekspresijskem ter nato v represijskem gojišču, ob ponovni kultivaciji v ekspresijskem gojišču po 8 urah kultivacije boljše pripravljena na agregacijski stres. To je razvidno kot povišana celična metabolna aktivnost ter zmanjšanje izražanja gena TDP-43 za polovico v primerjavi z ekspresijsko kulturo prve kultivacije. Naši rezultati bi lahko pomembno pripomogli k razumevanju razvoja ALS pri nastanku in odstranitvi agregatov proteina TDP-43.

• Language: Slovenian
• Keywords: Saccharomyces cerevisiae, TDP-43, ALS, agregati, stres
• Work type: Master's thesis/paper
• Typology: 2.09 - Master's Thesis
• Organization: BF - Biotechnical Faculty
• Publisher: [B. Jug]
• Year: 2020
• PID: 20.500.12556/RUL-114721
• UDC: 602.3:582.282.23:606:616.8(043.2)
• COBISS.SI-ID: 9436537
• Publication date in RUL: 06.03.2020

JUG, Blaž, 2020, Odziv kvasovk vrste Saccharomyces cerevisiae na tvorbo agregatov proteina TDP-43. [online]. Master’s thesis. B. Jug. [Accessed 18 May 2025]. Retrieved from: https://repozitorij.uni-lj.si/IzpisGradiva.php?lang=eng&id=114721

Secondary language

• Language: English
• Title: Response of yeast Saccharomyces cerevisiae to formation of TDP-43 protein aggregates

Abstract

Yeast Saccharomyces cerevisiae play an important role as a model organism for understanding the development of various neurodegenerative diseases. Their ease of cultivation and sufficient resemblance to mammalian cells have contributed to their general use in various scientific studies. Neurodegenerative diseases are a problem and a challenge of modern times, as they are the result of an aging population and the extended expectancy of human life. Amyotrophic lateral sclerosis (ALS) is a rare neurodegenerative disease. It affects motor neurons in the brain and spinal cord and causes their degeneration with cytoplasmic aggregates of the TDP-43 protein. The aim of the thesis was to study the yeast response to the presence of cytotoxic aggregates of protein TDP-43-GFP. Yeast were grown in successive batch cultivations and were observed using a fluorescence microscope at different stages of growth. At the same time, fluorescence intensity was measured to indicate the expression of the inserted human TDP-43-GFP gene, biomass yield for the effect of aggregates on cell growth and reproduction, and cellular metabolic activity as a reflection of energy needs for cellular stress response to the presence of aggregates. Experiments have revealed the stress effect of aggregates on cells and their response. Interestingly, the culture initially cultured in expression and then in the repression medium and yet again re-cultured in the expression medium was better prepared for aggregation stress after 8 hours of cultivation, which was reflected in lower gene expression intensity, higher biomass yield, and increased initial cellular metabolism. Our results could significantly help to understand the development of ALS in the formation and removal of TDP-43 protein aggregates.

• Keywords: Saccharomyces cerevisiae, TDP-43, ALS, aggregates, stress