Djelovanje membranskog transportera Oct1 u embrijima zebrice (Danio rerio)

master's thesis

• Title (english): Function od membrane transporter Oct1 in zebrafish embryos (Danio rerio)
• Author: Marina Horvat
• Mentor: Ivan Mihaljević (mentor)
• Mentor: Inga Urlić (sumentor)
• Committee member: Inga Urlić (predsjednik povjerenstva)
• Committee member: Ivana Šola (član povjerenstva)
• Committee member: Marin Ježić (član povjerenstva)
• Committee member: Romana Gračan (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2021-09-28, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Membranski transporteri ključni su za održavanje homeostaze i normalnog metabolizma, posebice u vrijeme ranog razvoja organizma. Transporteri organskih kationa (OCT, eng. Organic Cation Transporter), pripadnici superporodice SLC (eng. Solute Carrier), većinom su lokalizirani na membranama stanica bubrega i jetre, organima ključnim za biotransformaciju i izlučivanje ksenobiotika. Cilj ovog rada bio je istražiti djelovanje membranskih transportera embrija i ličinki zebrice (Danio rerio), s naglaskom na membranski transporter Oct1 iz porodice Slc22. Uz zebrice divljeg tipa, korištene su i one deficijentne za transporter Oct1, dobivene uspostavljenjem i genotipizacijom generacijskih linija genetički modificiranih mutanata. Praćenjem akumulacije fluorescentnih spojeva u zebricama in vivo pratilo se djelovanje membranskih transportera – unos u organizam, akumulacija u organima te izbacivanje u vanjski okoliš, dok se korištenjem supstrata transportera Oct1 pratilo njegovo djelovanje. Uz te metode in vivo, djelovanje transportera Oct1 istraženo je i imunofluorescencijskim analizama unutarstanične i tkivne ekspresije in vitro te analizom ekspresijskih profila membranskih transportera zebrica po embrionalnim i ličinačkim stadijima in silico. Rezultatima je pokazano da se transporter Oct1 nalazi u bubregu ličinke zebrice, gdje unosi spoj ASP+. Zaključeno je i da Oct1 ne utječe na vijabilnost i fertilnost zebrica promatrane starosti. Korištenjem zebrice tijekom ovih istraživanja pokazane su i njezine prednosti kao modelnog organizma kralježnjaka za genetička i ekotoksikološka istraživanja.

Abstract (english)

Membrane transporters are crucial for mantaining homeostasis and metabolism, especially during the early development of an organism. Organic Cation Transporters (OCTs), members of the Solute Carrier (SLC) superfamily, are mostly localized on the cell membranes of kidney and liver, organs crucial for biotransformation and xenobiotic secretion. The aim of this study was to investigate the activity of membrane transporters of zebrafish (Danio rerio) embryos and larvae, with emphasis on the membrane transporter Oct1, member of the Slc22 family. In addition to wild-type zebrafish, those deficient for the Oct1 transporter, obtained by establishing and genotyping generation lines of genetically modified mutants, were also used. The activity of membrane transporters was observed by in vivo monitoring of the accumulation of fluorescent compounds in zebrafish – their uptake into the organism, accumulation in organs and excretion into the environment, while the activity of Oct1 transporter was monitored by using its supstrates. In addition to these in vivo methods, the activity of the Oct1 transporter was investigated also by in vitro immunofluorescence analyzes of intracellular and tissue expression, and in silico analysis of the expression profiles of membrane transporters by embryonic and larval stages. The results show that the Oct1 transporter is located in the kidney of the zebrafish larva, where it transports ASP+. It was also concluded that Oct1 did not affect the viability and fertility of zebrafish of the observed ages. Using zebrafish throughout all experiments, benefits of zebrafish as a model vertebrate organism for genetic and ecotoxicological research have been demonstrated.

• Keywords: membranski transporteri; Oct1; zebrica; metabolizam; fluorescentne boje
• Keywords (english): membrane transporters; Oct1; zebrafish; metabolism; fluorescent dyes
• Language: croatian
• URN:NBN: urn:nbn:hr:217:710656
• Study programme: Title: Molecular Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access; Embargo expiration date: 2023-09-28
• Created on: 2021-10-08 10:21:28