MilliporeSigma
  • Mitophagy Failure in Fibroblasts and iPSC-Derived Neurons of Alzheimer's Disease-Associated Presenilin 1 Mutation.

Mitophagy Failure in Fibroblasts and iPSC-Derived Neurons of Alzheimer's Disease-Associated Presenilin 1 Mutation.

Frontiers in molecular neuroscience (2017-09-30)
Patricia Martín-Maestro, Ricardo Gargini, Andrew A Sproul, Esther García, Luis C Antón, Scott Noggle, Ottavio Arancio, Jesús Avila, Vega García-Escudero
ABSTRACT

Familial Alzheimer's disease (FAD) is clearly related with the accumulation of amyloid-beta (Aβ) and its deleterious effect on mitochondrial function is well established. Anomalies in autophagy have also been described in these patients. In the present work, functional analyses have been performed to study mitochondrial recycling process in patient-derived fibroblasts and neurons from induced pluripotent stem cells harboring the presenilin 1 mutation A246E. Mitophagy impairment was observed due to a diminished autophagy degradation phase associated with lysosomal anomalies, thus causing the accumulation of dysfunctional mitochondria labeled by Parkin RBR E3 ubiquitin protein ligase (PARK2). The failure of mitochondrial recycling by autophagy was enhanced in the patient-derived neuronal model. Our previous studies have demonstrated similar mitophagy impairment in sporadic Alzheimer's disease (AD); therefore, our data indicate that mitophagy deficiency should be considered a common nexus between familial and sporadic cases of the disease.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-Amyloid Precursor Protein, C-Terminal antibody produced in rabbit, IgG fraction of antiserum, buffered aqueous solution
Sigma-Aldrich
Carbonyl cyanide 3-chlorophenylhydrazone, ≥97% (TLC), powder
Sigma-Aldrich
Monoclonal Anti-β-Actin antibody produced in mouse, clone AC-15, ascites fluid
Sigma-Aldrich
Anti-Tau-1 Antibody, clone PC1C6, clone PC1C6, Chemicon®, from mouse
Sigma-Aldrich
Anti-Tau Mouse mAb (TAU-5), liquid, clone TAU-5, Calbiochem®
Sigma-Aldrich
Anti-TRA-1-81 Antibody, clone TRA-1-81, clone TRA-1-81, Chemicon®, from mouse
Sigma-Aldrich
Laminin from Engelbreth-Holm-Swarm murine sarcoma basement membrane, 1-2 mg/mL in Tris-buffered saline, 0.2 μm filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
Poly-L-ornithine hydrobromide, mol wt 30,000-70,000
Sigma-Aldrich
Monoclonal Anti-β-Tubulin antibody produced in mouse, clone TUB 2.1, ascites fluid
Sigma-Aldrich
Anti-BNIP3 antibody, Mouse monoclonal, clone ANa40, purified from hybridoma cell culture
Sigma-Aldrich
Monoclonal Anti-MAP2 antibody produced in mouse, clone HM-2, ascites fluid
Sigma-Aldrich
Anti-NeuN Antibody (rabbit), from rabbit, purified by affinity chromatography
Sigma-Aldrich
Anti-Vesicular Glutamate Transporter 1 Antibody, serum, Chemicon®
Sigma-Aldrich
Anti-TRA-1-60 Antibody, clone TRA-1-60, clone TRA-1-60, Chemicon®, from mouse