Detection of stem cells in the brain of genetically modified mouse B4Galnt1

Uvod: Neurogeneza, odnosno stvaranje novih živčanih stanica, osim u embrionalnom razvoju, odvija se i u odraslom mozgu i to uglavnom u područjima: u subventrikularnoj zoni (SVZ) i subgranularnoj zoni dentatnog girusa (DG) hipokampusa te u njušnoj lukovici. U tom procesu ulogu imaju i kompleksni gangliozidi koji su prisutni u svim tkivima, ali su najzastupljeniji u mozgu, a njihov nedostatak dovodi do neurodegeneracije i neuroloških poremećaja. Ciljevi: Odrediti povezanost nedostatka kompleksnih gangliozida i biljega upale i proliferacije stanica. Materijali i metode: U istraživanje su uključena dva tipa miša - genetički modificirani B4Galnt1 (KO) s nedostatkom u sintezi kompleksnih gangliozida i kontrolni miš divljeg tipa (WT). Imunohistokemijskom analizom pomoću protutijela Iba1, GFAP i PCNA detektirani su navedeni biljezi. Rezultati: Rezultati su analizirani računalnim programom Fiji te su statistički obrađeni. Dobiveni su rezultati pokazali statistički značajnu razliku kod sva tri biljega. U SVZ-u je uočeno da je Iba1 više izražen kod KO-a u odnosu na WT, dok su vrijednosti PCNA biljega bile veće u WT-u uspoređujući ga s KO mišem. S druge strane, u DG-u je pokazana veća ekspresija GFAP pozitivnih stanica u WT mišu nego u KO. Zaključak: Veće vrijednosti biljega Iba1 potvrđuju hipotezu o povećanom broju mikroglija stanica pri upalnom procesu, dok manje vrijednosti PCNA u KO mišu upućuju na moguće postojanje smanjenog ukupnog broja matičnih stanica u SVZ-u. Slično, manji broj GFAP pozitivnih stanica u hipokampusu KO miša moglo bi se povezati s njihovom slabijom mogućnošću diferencijacije za vrijeme upale.Introduction: Neurogenesis, or the creation of new neuron cells, occurs not only in embryonic development, but also in the adult brain, mainly in two regions: the subventricular zone (SVZ) and the subgranular zone of the dentate gyrus (DG) of the hippocampus and also in olfactory bulbus. Complex gangliosides are also involved in this process and they are present in all tissues, but are the most prominent in the brain, and their deficiency leads to neurodegeneration and neurological disorders. Goals: This study investigates the correlation between lack of complex gangliosides, inflammation and cell proliferation. Materials and methods: Two types of mice have been included in the study - genetically modified B4Galnt1 (KO) with a lack of complex ganglioside synthesis and a control wild type mouse (WT). Immunohistochemical analysis by means of antibodies Iba1, GFAP and PCNA detects the aforementioned markers. Results: The results were analysed by a computer program Fiji and statistically processed. The results obtained indicated statistically significant difference in all three markers. In SVZ it was found that Iba1 was more expressed in KO compared to WT, while PCNA markers were higher in WT compared with KO mice. On the other hand, DG showed greater expression of GFAP positive cells in WT mouse than in KO. Conclusion: Higher values for Iba1 marker confirms the hypothesis of the increased number of microglia cells in the inflammatory process, while the lower PCNA values in KO mouse indicate the possible existence of a reduced total stem cell count in the SVZ. Similarly, a smaller number of GFAP positive cells in the KO mouse's hippocampus may be related to their lower differentiation ability during inflammation