Priprema i karakterizacija vezikula plazmatske membrane iz stanica alga

master's thesis

• Title (english): Preparation and characterization of algal plasma membrane vesicles
• Author: Tomo Balen
• Mentor: Nadica Ivošević DeNardis (mentor)
• Committee member: Iva Juranović Cindrić (predsjednik povjerenstva)
• Committee member: Đani Škalamera (član povjerenstva)
• Committee member: Jana Pisk (član povjerenstva)
• Committee member: Sanda Rončević (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Chemistry); Zagreb
• Defense date and country: 2022-02-25, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Chemistry

Abstract

U ovom je radu provedena priprema i karakterizacija vezikula plazmatskih membrana
dobivenih iz mikroalga Dunaliella tertiolecta uzgojenih kod dvaju saliniteta. Osmotskim
šokom mekanih stanica nastale su mikrometarske vezikule koje su karakterizirane metodama
oslikavanja primjenom epifluorescencijske, konfokalne i mikroskopije atomskih sila.
Pripremljene vezikule bile su mikrometarske, pojedinačne, uglavnom sferičnoga oblika, prazne
ili s nešto staničnoga materijala te su pokazale svojstvo autofluorescencije uslijed prisutnosti
klorofila koji je ugrađen tijekom rekonstrukcije membrane. Oslikavanjem na nanometarskoj
skali vizualizirane su površinske globularne strukture koje bi mogle odgovarati glikoproteinima
između kojih su nanometarske pore. Vezikule plazma membrane su propusne za male hidrofilne
molekule kalceina, dok su polupropusne za veće molekule konkanavalin A-FITC uz specifičnu
interakciju s površinskim skupinama glukozil i manozil na membrani. Rezultati nisu pokazali
značajnu razliku u smislu morfologije i propusnosti između vezikula plazma membrana kod
dva saliniteta. Rezultati ovoga rada doprinose daljnjem razvoju nove generacije alternativnih
bioinspiriranih nosača aktivnih komponenti.

Abstract (english)

In this Thesis plasma membrane vesicles of Dunaliella tertiolecta microalgae grown at two
salinities were prepared and characterized. Osmotic shock of soft cells resulted in micrometersized vesicles that were characterized by imaging methods using fluorescence, confocal, and
atomic force microscopy. The prepared vesicles were micrometer-sized, single, mostly
spherical, empty or with some cellular material, and exhibited the property of autofluorescence
due to the presence of chlorophyll incorporated during membrane reconstruction. Nanometerscale measurements show spherical surface structures that could correspond to glycoproteins,
with nanometer pores in between. Plasma membrane vesicles are permeable to small
hydrophilic calcein molecules, whereas they are semipermeable to larger concavalin A-FITC
molecules consistent with specific interaction with glucosyl and mannosyl groups on the
membrane. The results showed no significant difference in morphology and permeability
between plasma membrane vesicles in the two salinities. The results of this work contribute to
the further development of a new generation of alternative bioinspired drug carriers.

• Keywords: Dunaliella tertiolecta; fluorescentne boje; konfokalna mikroskopija; mikroskopija atomskih sila; nanomorfologija; propusnost membrane; vezikule plazmatske membrane
• Keywords (english): atomic force microscopy; confocal microscopy; Dunaliella tertiolecta; fluorescent dye; membrane permeability; nanomorphology; plasma membrane vesicles
• Language: croatian
• URN:NBN: urn:nbn:hr:217:852292
• Project: Number: IP-2018-01-5840; Title: Od površinskih svojstava stanica alga do pokazatelja stresa u vodenim ekosustavima; Acronym: CELLSTRESS; Leader: Nadica Ivošević DeNardis; Jurisdiction: Croatia; Funder: HRZZ; Funding stream: IP
• Study programme: Title: Graduate university studies in chemistry; specializations in: Research programme; Course: Research programme; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra kemije (magistar/magistra kemije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access; Embargo expiration date: 2024-02-25
• Created on: 2022-03-02 13:28:56