Inhibition of PIKfyve by YM-201636 dysregulates autophagy and leads to apoptosis-independent neuronal cell death

Sally Martin; Callista B Harper; Linda M May; Elizabeth J Coulson; Frederic A Meunier; Shona L Osborne

doi:10.1371/journal.pone.0060152

Inhibition of PIKfyve by YM-201636 dysregulates autophagy and leads to apoptosis-independent neuronal cell death

PLoS One. 2013;8(3):e60152. doi: 10.1371/journal.pone.0060152. Epub 2013 Mar 27.

Authors

Sally Martin¹, Callista B Harper, Linda M May, Elizabeth J Coulson, Frederic A Meunier, Shona L Osborne

Affiliation

¹ The University of Queensland, Queensland Brain Institute, Brisbane, Queensland, Australia.

Abstract

The lipid phosphatidylinositol 3,5-bisphosphate (PtdIns(3,5)P 2), synthesised by PIKfyve, regulates a number of intracellular membrane trafficking pathways. Genetic alteration of the PIKfyve complex, leading to even a mild reduction in PtdIns(3,5)P 2, results in marked neurodegeneration via an uncharacterised mechanism. In the present study we have shown that selectively inhibiting PIKfyve activity, using YM-201636, significantly reduces the survival of primary mouse hippocampal neurons in culture. YM-201636 treatment promoted vacuolation of endolysosomal membranes followed by apoptosis-independent cell death. Many vacuoles contained intravacuolar membranes and inclusions reminiscent of autolysosomes. Accordingly, YM-201636 treatment increased the level of the autophagosomal marker protein LC3-II, an effect that was potentiated by inhibition of lysosomal proteases, suggesting that alterations in autophagy could be a contributing factor to neuronal cell death.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Aminopyridines / pharmacology*
Animals
Apoptosis / drug effects*
Autophagy
Endocytosis / drug effects
Endosomes / drug effects
Endosomes / metabolism
Heterocyclic Compounds, 3-Ring / pharmacology*
Hippocampus / cytology
Immunohistochemistry
Lysosomes / drug effects
Lysosomes / metabolism
Mice
Mice, Inbred C57BL
Neuroendocrine Cells / cytology
Neuroendocrine Cells / drug effects
Neurons / cytology*
Neurons / drug effects
Neurons / metabolism
Neurons / ultrastructure
PC12 Cells
Phosphatidylinositol 3-Kinases / metabolism*
Phosphoinositide-3 Kinase Inhibitors
Protein Transport / drug effects
Rats
Vacuoles / drug effects
Vacuoles / metabolism

Substances

6-amino-N-(3-(4-(4-morpholinyl)pyrido(3',2'-4,5)furo(3,2-d)pyrimidin-2-yl)phenyl)-3-pyridinecarboxamide
Aminopyridines
Heterocyclic Compounds, 3-Ring
Phosphoinositide-3 Kinase Inhibitors
Pikfyve protein, mouse

Grants and funding

This work was supported by the National Health and Medical Research Council Australia (S.L.O grant number 631418). F.A.M. and E.J.C. are Research Fellows of the National Health and Medical Research Council Australia. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.