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P2‐262: Apoptosis inducing factor and lipoxygenases in mitochondrial function affected by beta‐amyloid peptides
Author(s) -
Cieslik Magdalena,
Strosznajder Robert P.,
Pajak Beata,
Gajkowska Barbara,
Strosznajder Joanna B.
Publication year - 2010
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.2010.05.1312
Subject(s) - mitochondrion , oxidative stress , apoptosis , microbiology and biotechnology , poly adp ribose polymerase , apoptosis inducing factor , cytosol , biochemistry , programmed cell death , nitric oxide , chemistry , amyloid (mycology) , biology , enzyme , caspase , polymerase , endocrinology , inorganic chemistry
Background: Oxidative stress occurs early in the progression of Alzheimer’s disease (AD). Non-enzymatic modifications induced by oxidative stress may also arise from reaction with low Mw reactive carbonyl compounds such as glyoxal (GO), methylglyoxal (MGO) and malondialdehyde (MDA), resulting from damaged carbohydrates or unsaturated fatty acids leading to the formation of both adducts and cross-links denominated Advanced Glycation/Lipoxidation End products (AGE/ALEs). It has been described that pro-nerve growth factor (pro-NGF) has increased levels in Alzheimer’s Disease (AD) affected brains and is able to induce cell death in cultures, however the reason for these phenomena is still elusive. Methods: Post-mortem human brain samples of AD and control cases were obtained from the Institute of Neuropathology Brain Bank. Antibody against pro-NGF prodomain was made as described Beattie et al. Secondary antibodies (anti-mouse IgG-HRP and anti-rabbit IgG-HRP) were obtained from Amersham Biosciences (Piscataway, NJ). CEL, CML and AGEs specific monoclonal antibodies were purchased from TransGenic Inc., (Kobe). Protein immunodetection, Pro-NGF immunoprecipitation; expression, purification and refolding of pro-NGF from Escheria coli, modifications of rh-pro-NGF, isolation of Pro-NGF from human brain, cell cultures and treatments, detection of apoptosis in the cell cultures, pro-NGF in vivo administration, and behavioral tests were done. Results: We show that pro-NGF in human Alzheimer’s disease-affected hippocampus and entorhinal cortex is modified by advanced glycation and lipoxidation end-products (AGE/ ALEs) in a stage-dependent manner. These modifications block the proNGF processing to mature NGF (NGF), thus making the proneurotrophin especially effective in inducing apoptosis of PC12 cells in culture through p75NTR. The processing of AGE/ALE in vitro modified recombinant human pro-NGF (rh-pro-NGF) is severely impaired as evidenced by Western blot and by examining its physiological functionality in cell cultures. We also report that modified rh-pro-NGF as well as pro-NGF isolated from human brain affected by AD, cause impairment of learning tasks when administered intracerebroventricularly in mice that correlates with Alzheimer’s disease-associated learning impairment. Conclusions: The data we present here offer a novel pathway of ethiopathogenesis in Alzheimer’s disease caused by AGE/ALEs modification of pro-NGF.