Utjecaj unosa virusnog vektora u mišji mozak na aktivaciju mikroglije

master's thesis

• Title (english): The effect of viral vector introduction into mouse brain on activation of the microglia
• Author: Andrea Belamarić
• Mentor: Srećko Gajović (mentor)
• Mentor: Aleksandar Vojta (komentor)
• Committee member: Aleksandar Vojta (predsjednik povjerenstva)
• Committee member: Sandra Hudina (član povjerenstva)
• Committee member: Biljana Balen (član povjerenstva)
• Committee member: Tomislav Ivanković (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2023-09-26, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Virusni vektori predstavljaju efikasan alat za unos genetičkog materijala u stanice te njihova
primjena ima važnu ulogu u raznim terapijama i neuroznanstvenim istraživanjima. Postoje
različite metode unosa virusnog vektora u središnji živčani sustav, a jedna od njih je direktno
stereotaksijsko injektiranje. Budući da mikroglija ima ulogu nadzornika u mozgu te sudjeluje
u aktivaciji imunosnog odgovora, cilj ovog rada bio je ispitati uzrokuje li stereotaksijsko
injektiranje lentivirusnog vektora aktivaciju upalnog odgovora (tj. mikroglije), promjenu u
morfologiji mikroglije te promjenu u ekspresiji TLR2 receptora koji se aktivira u slučaju
infekcije. Metoda imunohistokemijskog bojenja provedena je 7. i 14. dan nakon injektiranja
na tankim prerezima mišjih mozgova u svrhu vizualizacije markera mikroglije (Iba-1) i TLR2.
Upalni odgovor je lokaliziran u području prolaska igle, prolazan i popraćen promjenom u
morfologiji mikroglije te je značajno manji 14. dan u odnosu na 7. dan nakon unosa. U
području injektiranja utvrđena je povećana ekspresija TLR2 7. dan nakon unosa, no 14. dan je
značajno smanjena. Ovo istraživanje ukazuje kako za aktivaciju upalnog odgovora u mozgu
nije odgovoran lentivirusni vektor, već metoda unosa (stereotaksijsko injektiranje). Također,
istraživanjem je potvrđeno kako lentivirusni vektori predstavljaju siguran alat za unos
genetičkog materijala u mišji mozak.

Abstract (english)

Viral vectors represent an efficient tool for introducing genetic material into cells and their
application plays an important role in various therapies and neuroscientific research. Different
methods can be used to introduce the viral vector into the central nervous system, and one of
them is direct stereotaxic injection. Since microglia are considered the main supervisors in the
brain and participate in the inflammatory response activation, the aim of this work was to
examine whether the stereotaxic injection of a lentiviral vector causes the activation of an
inflammatory response (i.e. microglia), change in the microglia morphology and change in the
expression of TLR2, a receptor activated in case of infection. The immunohistochemical
staining method was performed 7 and 14 days post injection on thin sections of mice brains in
order to visualize the microglia marker (Iba-1) and TLR2. The inflammatory response is
localized in the injection site, is transient, accompanied by a change in the microglia
morphology and significantly lower 14 days post injection in comparison to 7 days post
injection. In the injection site, an increased TLR2 expression was also recorded 7 days post
injection, while 14 days post injection it was significanly lower. This research established that
the activation of inflammatory response in the brain is not caused by the lentiviral vector, but
the introduction method (stereotaxic injection). Moreover, this research confirmed that
lentiviral vectors represent a safe tool for introducing genetic material into the mouse brain.

• Keywords: upalni odgovor; Iba-1; TLR2; imunohistokemija
• Keywords (english): inflammatory response; Iba-1; TLR2; immunohistochemistry
• Language: croatian
• URN:NBN: urn:nbn:hr:217:560020
• 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: Embargoed access; Embargo expiration date: 2025-09-26
• Created on: 2023-10-16 10:14:15