Imunokemijska karakterizacija protutijela za membranske proteine OATP1D1 i OCT1 u uzorcima zebrice (Danio rerio)

master's thesis

• Title (english): Immunochemical characterization of antibodies for membrane proteins OATP1D1 and OCT1 in zebra samples (Danio rerio)
• Author: Ana Dananić
• Mentor: Bojan Žunar (mentor)
• Mentor: Dean Karaica (komentor)
• Committee member: Igor Stuparević (predsjednik povjerenstva)
• Committee member: Jovica Lončar (član povjerenstva)
• Committee member: Igor Slivac (član povjerenstva)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Chemistry and Biochemistry); (Laboratory for Biochemistry); Zagreb
• Defense date and country: 2023-09-06, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Prijenosnici otopljenih tvari (engl. Solute Carriers, SLC) velika su skupina membranskih proteina koji igraju ključnu ulogu u uklanjanju različitih endogenih i egzogenih molekula iz organizma zbog čega je njihova ekspresija u membrani stanica u određenim organima neophodna kod svih eukariota. SLC21 i SLC22, članovi superobitelji SLC, dvije su važne obitelji prijenosnika s čijim proteinima ćemo se susresti u ovom radu, a čija je lokalizacija kod ljudi relativno dobro istražena. Međutim, sve je veći interes za razumijevanjem njihove lokalizacije i u drugim animalnim modelima, poput ribe zebrice. Stoga, cilj ovog istraživanja je bio, korištenjem imunokemijskih metoda i knock-out modela zebrice, potvrditi specifičnost vezanja dizajniranih zečjih poliklonskih protutijela za membranske proteine Oatp1d1 iz obitelji Slc21 i Oct1 iz obitelji Slc22 zebrice, a zatim i otkriti/opisati lokalizaciju istih proteina u embrijima zebrica i tkivima odraslih jedinki. Rezultati navedenih analiza potvrdili su specifično vezanje dizajniranih protutijela za protein Oatp1d1 u membrani hepatocita jetre, kao i specifično vezanje nekih poliklonskih protutijela za protein Oct1, lokaliziran u bazolateralnoj membrani bubrega odraslih jedinki.

Abstract (english)

Solute carriers (SLC) are a large group of membrane proteins that play a key role in the removal of various endogenous and exogenous molecules from the organism, which is why their expression in the cell membrane of certain organs is necessary in all eukaryotes. SLC21 and SLC22, members of the SLC superfamily, are two important families of transporters whose proteins we will mention in this work, and whose localization in humans has been relatively well investigated. However, there is a growing interest in understanding their localization in other animal models, such as zebrafish. Therefore, the goal of this research was, to confirm the binding specificity of the designed rabbit polyclonal antibodies to the membrane proteins Oatp1d1 from the family Slc21 and Oct1 from the family Slc22 in zebrafish, and then to discover/describe the localization of the same proteins in zebrafish embryos and tissues of adults. The results of aforementioned analyses confirmed the specific binding of the designed antibodies to the protein Oatp1d1 in the liver hepatocyte membrane, as well as the specific binding of some polyclonal antibodies to the protein Oct1, localized in the basolateral membrane of the kidney in adults individuals.

• Keywords: Danio rerio; imunokemijska karakterizacija; knock-out; kriosekcije zebrice; fluorescentna mikroskopija
• Keywords (english): Danio rerio; immunochemical characterization; knock-out; zebrafish cryosections; fluorescence microscopy
• Language: croatian
• URN:NBN: urn:nbn:hr:159:610799
• Study programme: Title: Molecular Biotechnology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra inženjer/inženjerka molekularne biotehnologije (magistar/magistra inženjer/inženjerka molekularne biotehnologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Closed access
• Created on: 2024-06-27 09:32:40