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master's thesis
Analiza supstrata enzima BACE1 u endosomalnim frakcijama moždanih regija miševa NPC1-/- i divljeg tipa miša (NCP1+/+)
Zagreb: University of Zagreb, Faculty of Science, 2021. urn:nbn:hr:217:819821
Meglaj, Sarah
University of Zagreb
Faculty of Science
Department of Biology

Institutional repository: Repository of the Faculty of Science

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Meglaj, S. (2021). Analiza supstrata enzima BACE1 u endosomalnim frakcijama moždanih regija miševa NPC1-/- i divljeg tipa miša (NCP1+/+) (Master's thesis). Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:819821

Meglaj, Sarah. "Analiza supstrata enzima BACE1 u endosomalnim frakcijama moždanih regija miševa NPC1-/- i divljeg tipa miša (NCP1+/+)." Master's thesis, University of Zagreb, Faculty of Science, 2021. https://urn.nsk.hr/urn:nbn:hr:217:819821

Meglaj, Sarah. "Analiza supstrata enzima BACE1 u endosomalnim frakcijama moždanih regija miševa NPC1-/- i divljeg tipa miša (NCP1+/+)." Master's thesis, University of Zagreb, Faculty of Science, 2021. https://urn.nsk.hr/urn:nbn:hr:217:819821

Meglaj, S. (2021). 'Analiza supstrata enzima BACE1 u endosomalnim frakcijama moždanih regija miševa NPC1-/- i divljeg tipa miša (NCP1+/+)', Master's thesis, University of Zagreb, Faculty of Science, accessed 25 August 2024, https://urn.nsk.hr/urn:nbn:hr:217:819821

Meglaj S. Analiza supstrata enzima BACE1 u endosomalnim frakcijama moždanih regija miševa NPC1-/- i divljeg tipa miša (NCP1+/+) [Master's thesis]. Zagreb: University of Zagreb, Faculty of Science; 2021 [cited 2024 August 25] Available at: https://urn.nsk.hr/urn:nbn:hr:217:819821

S. Meglaj, "Analiza supstrata enzima BACE1 u endosomalnim frakcijama moždanih regija miševa NPC1-/- i divljeg tipa miša (NCP1+/+)", Master's thesis, University of Zagreb, Faculty of Science, Zagreb, 2021. Available at: https://urn.nsk.hr/urn:nbn:hr:217:819821

Metadata
TitleAnaliza supstrata enzima BACE1 u endosomalnim frakcijama moždanih regija miševa NPC1-/- i divljeg tipa miša (NCP1+/+)
Title (english)Distribution of BACE1 substrates in the endosomal fractions isolated from the brain regions of wild type (NCP1+/+) and NPC1-/- disease mouse model
AuthorSarah Meglaj
MentorSilva Katušić Hećimović (mentor)
MentorJulija Erhardt (sumentor)
Committee memberJulija Erhardt (predsjednik povjerenstva)
Committee memberJasna Lajtner (član povjerenstva)
Committee memberBiljana Balen (član povjerenstva)
Committee memberRomana Gračan (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Science
(Department of Biology)
Zagreb
Defense date and country2021-07-08, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Biology
Abstract
Niemann-Pickova bolest tipa C (NPC) nasljedna je, rijetka neurodegenerativna bolest nakupljanja lipida uzrokovana mutacijama u genu NPC1, a rjeđe NPC2. NPC dijeli niz karakteristika s kompleksnom Alzheimerovom bolesti (AB), ukazujući na postojanje zajedničkih patoloških procesa. Neke od zajedničkih karakteristika su disfunkcija endolizosomalnog puta te nedavno otkrivena povećana proteoliza supstrata β-sekretaze (BACE1) – enzima ključnog u patogenezi AB. Cilj rada bio je ispitati hipotezu da je povećana proteoliza supstrata BACE1 u moždanim regijama miševa NPC1 (NPC1-/-) u odnosu na miševe divljeg tipa (WT, NPC1+/+) posljedica nakupljanja supstrata i/ili enzima BACE1 u endosomima – organelima odgovornima za cijepanje BACE1. Metodom ultracentrifugiranja u diskontinuiranom gradijentu saharoze, izdvojene su frakcije ranih endosoma te kasnih endosoma/lizosoma iz malog mozga i hipokampusa devet tjedana starih miševa NPC1 i WT te su analizirane metodom western blotting, a dobiveni signali kvantificirani su softwareom ImageJ. Rezultati pokazuju različitu distribuciju i razinu biljega ranih endosoma i kasnih endosoma/lizosoma u moždanim regijama NPC1 u odnosu na WT miševe. Također, uočeno je značajno nakupljanje BACE1 supstrata Sez6L u ranim endosomima moždanih regija miševa NPC1 vs. WT. Zaključno, rezultati ukazuju da je Alzheimeru sličan fenotip u bolesti NPC jednim dijelom vjerojatno posljedica poremećaja transporta u endolizosomalnom putu te nakupljanja i povećanog cijepanja supstrata BACE1 u endosomima.
Abstract (english)
Niemann-Pick type C disease (NPC) is a rare, inherited, neurodegenerative lipid storage disorder caused by mutations in NPC1 or NPC2 gene. NPC shares several features with the complex Alzheimer's disease (AD), including endolysosomal dysfunction and the increased proteolysis of the substrates of β-secretase (BACE1), a key protease in AD pathogenesis. The goal of this work was to test the hypothesis that increased proteolysis of BACE1 substrates in brain regions of NPC1 (NPC1-/-) compared to wild-type mice (WT, NPC1+/+) is due to accumulation of BACE1 substrates and/or enzyme BACE1 in endosomes. Ultracentrifugation in a discontinuous sucrose gradient was used to isolate fractions of early and late endosomes/lysosomes from the cerebellum and hippocampus of nine-week-old NPC1 and WT mice. The fractions were analyzed by western blotting, and the obtained signals were quantified using ImageJ software. Different distribution and level of markers of early and late endosomes/lysosomes in the brain regions of NPC1 vs. WT mice was observed. Also, significant accumulation of BACE1 substrate Sez6L was detected in early endosomes in brains of NPC1 mice. In conclusion, the results suggest that the Alzheimer-like phenotype in NPC disease is likely due to endolysosomal dysfunction causing accumulation and increased cleavage of BACE1 substrates in endosomes.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:217:819821
Study programmeTitle: Experimental Biology
Study programme type: university
Study level: graduate
Academic / professional title: magistar/magistra eksperimentalne biologije (magistar/magistra eksperimentalne biologije)
Type of resourceText
File originBorn digital
Access conditionsOpen access
Embargo expiration date: 2023-07-08
Terms of useLicence In copyright icon
Created on2021-07-22 12:12:06