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P1‐179: A CORRELATION ANALYSIS BETWEEN CLINICAL AND NEUROPATHOLOGICAL DATA FROM FAMILIAL ALZHEIMER DISEASE CASES WITH MISSENSE MUTATIONS OF APP AND THE FEATURES OF MUTANT APP PROCESSING IN CELL MODELS
Author(s) -
Bi Christopher,
Li Bin
Publication year - 2018
Publication title -
alzheimer's and dementia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 6.713
H-Index - 118
eISSN - 1552-5279
pISSN - 1552-5260
DOI - 10.1016/j.jalz.2018.06.183
Subject(s) - missense mutation , amyloid precursor protein , pathological , neuroinflammation , extracellular , mutation , disease , biology , tau protein , mutant , pathogenesis , alzheimer's disease , neuroscience , medicine , pathology , genetics , gene
mice over-expressing Nrf2 in astrocytes using the GFAP-promoter(GFAP-Nrf2)(Vargas et al. 2008). Ab-fractionation: Cortical and hippocampal tissues were homogenized in Triton X-100lysis buffer;lysates were centrifuged at 100.000xg/60min@4 C. Supernatant was collected(soluble fractions) and pellets were 1%-SDS-resuspended and centrifuged as above; supernatant collected(SDS-soluble fraction); the remaining pellets were dissolved in urea buffer(insoluble fraction). ELISA: Ten mg total lysates were assayed by hAb(1-42) ELISA(Invitrogen). Western Blot(WB): Soluble/insoluble fractions of Ab/APPs were resolved by 10%Tris-tricine; total lysates were resolved by 10%Trisglycine SDS-PAGE. Immunohistochemistry: Ten mm coronal paraffin-embedded sections were stained for Aß/APP using VECTASTAIN -ABC system. Immunofluorescence staining for other proteins used AlexaFluor -conjugated secondary antibodies. Results: Levels of Aß and APP detected by 6E10, 22C11, Aß(1-40)/Aß(1-42) immunostaining, demonstrated reduced staining intensity in plaques and in neurons in tissues from cortex and the CA-1 region of hippocampus from APPPS1/GFAP-Nrf2 mice compared with APP/PS1 mice.A significant reduction of Ab oligomers and APP N-terminal fragments in SDS-soluble and urea-insoluble fractions was observed by WB analysis comparing these same cohorts ELISA for hAb(142) showed similar and significant decreases; as did markers for inflammation and autophagy observed by WB and immunohistochemistry for GFAP, Iba-1, NeuN, LCIII and p62/SQSTM1. Conclusions: Astrocytic over-expression of Nrf2 in APP/PS1 mice have reduced disease progression when compared with APP/ PS1 mice. Nrf2 may play an important therapeutic role in APP processing and Ab degradation in AD.