Moderate activation of autophagy regulates the intracellular calcium ion concentration and mitochondrial membrane potential in beta-amyloid-treated PC12 cells

Neurosci Lett. 2016 Apr 8:618:50-57. doi: 10.1016/j.neulet.2016.02.044. Epub 2016 Feb 26.

Abstract

Alzheimer's disease (AD) is an age-related and progressive neurodegenerative disease. Aggregated beta-amyloid (Aβ) disturbs Ca(2+) homeostasis and causes mitochondrial dysfunction and finally underlies AD. Recent evidence suggests that autophagy initiation by Beclin-1 protein might be involved in the pathogenesis of AD. However, the effects of Beclin-1 dependent autophagy on intracellular calcium ion concentration ([Ca(2+)]i) and mitochondrial membrane potential (MMP) is unclear. The effects of Beclin-1 dependent autophagy that were activated by a gradient concentration of autophagy activator rapamycin or inhibited by autophagy inhibitor 3-methyladenine (3-MA) on cell viability and cell morphology were examined. Pretreatment with rapamycin significantly up-regulated the expression of Beclin-1 in response to Aβ1-42 application, but after pretreatment with 3-MA it was significantly down-regulated. Moderate activation of Beclin-1 dependent autophagy had an up regulation effect on cell viability and could maintain the original morphology of cells. Furthermore, rapamycin or 3-MA on [Ca(2+)]i and MMP in Aβ1-42 treatment of PC12 cells were evaluated. We also report that PC12 cells treated with Aβ1-42 showed an increase in [Ca(2+)]i but a decrease in MMP when compared to the normal control. However the application of rapamycin prior to this prevented the increase in [Ca(2+)]i and the decrease in MMP in response to Aβ1-42. When 3-MA was applied this exacerbated the effect of Aβ1-42 on the [Ca(2+)]i and the MMP. This shows that moderate activation of Beclin-1 dependent autophagy by rapamycin can modulate Ca(2+) homeostasis and maintain MMP in response to Aβ1-42 induced cytotoxicity and so may have a preventive function in AD.

Keywords: Alzheimer’s disease; Autophagy; Beclin-1; Beta-amyloid; Intracellular calcium; Mitochondrial membrane potential.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenine / analogs & derivatives
  • Adenine / pharmacology
  • Amyloid beta-Peptides / metabolism*
  • Amyloid beta-Peptides / toxicity
  • Animals
  • Apoptosis Regulatory Proteins / metabolism
  • Autophagy / drug effects*
  • Beclin-1
  • Calcium / metabolism*
  • Cell Survival / drug effects
  • Intracellular Space / metabolism
  • Membrane Potential, Mitochondrial*
  • Mitochondria / drug effects
  • Mitochondria / ultrastructure
  • PC12 Cells
  • Peptide Fragments / metabolism*
  • Peptide Fragments / toxicity
  • Phagosomes / drug effects
  • Phagosomes / ultrastructure
  • Rats
  • Sirolimus / pharmacology

Substances

  • Amyloid beta-Peptides
  • Apoptosis Regulatory Proteins
  • Beclin-1
  • Becn1 protein, mouse
  • Peptide Fragments
  • amyloid beta-protein (1-42)
  • 3-methyladenine
  • Adenine
  • Calcium
  • Sirolimus