Project description:Genome wide DNA methylation profiling of normal and APP/PSEN1 mice. A custom Illumina Golden Gate DNA methylation Beadchip was used to obtain DNA methylation profiles across approximately 800 CpGs.
Project description:Genome wide DNA methylation profiling of normal and APP/PSEN1 mice. A custom Illumina Golden Gate DNA methylation Beadchip was used to obtain DNA methylation profiles across approximately 800 CpGs. Bisulphite converted DNA from the 96 samples were hybridised to the Illumina custom golden gate DNA methylation array.
Project description:Microglia was isolated from Egln3–/–; App-Psen1 and App-Psen1 mice using flow cytometry to identify changes in their expression profile and phenotype associated with the absence of Prolyl Hydroxylase 3 (PHD3).
Project description:The purpose of this project was to compare whole genome expression in 5 transgenic mice with human genes for dementia that result in either plaques or tangle pathology to the expression in wild-type control mice and to each other at different stages of disease progression. Total RNA was obtained from hippocampus, cortex and cerebellum in four lines of ‘amyloid’ transgenic mice (mutant human APP and APP/PSEN1 genes) and ‘TAU’ transgenic mice (mutant human MAPT gene) as well as wild-type control mice at 8,16, 32 and 72 weeks
Project description:Dysfunctional mitophagy is considered an important component of Alzheimer's disease (AD) pathology, however, direct in vivo evidence and underlying mechanisms are lacking. Using a newly established AD mitophagy reporter mouse model (APP/PSEN1/mt-keima), we find that the basal mitophagy of neuronal soma in the hippocampal CA1 region of AD mice is selectively impaired. Pathologic mitophagy is highly activated in neurites in both the cortex and hippocampus regions of AD mice, with abnormal accumulation of mitochondria. These aberrantly accumulated mitochondria are fragmented and trigger massive formation of mitophagosomes and mitolysomes. However, the degradation of mitochondria is incomplete, likely due to inadequate maturation of lysosomal protease and de-acidification of lysosomes, leading to the mitochondrial accumulation within autophagic vacuoles. The resulting swollen neurites coalesce into structures containing mitochondria at different stages of degradation, that we term mitochondrial plaques (MPs). While MPs and Aβ plaques can emerge independently at early stages, they eventually develop into mixed MP/Aβ plaques. MPs are positive of amyloid precursor protein (APP) and contribute to over 60% of all APP detected in/around MPs, explaining the eventual development of mixed plaques. Our study identifies mitochondria as prominent constituent within autophagic vacuoles in dystrophic neurites and MPs as a promising new therapeutic target for AD. The hippocampus samples of APP/PSEN1/mt-keima and WT/mt-keima at 9, 15, 25, 30 and 50 wks were collected from 3 mice per age group per genotype group and subjected to total RNA isolation to understand what metabolism-related gene expression and pathways changes occur in AD during aging.
