Abstract
We have previously demonstrated that oligomeric amyloid β peptide (oAβ) together with iron overload generates synaptic injury and activation of several signaling cascades. In this work, we characterized hippocampal neuronal response to oAβ. HT22 neurons exposed to 500 nM oAβ showed neither increased lipid peroxidation nor altered mitochondrial function. In addition, biophysical studies showed that oAβ did not perturb the lipid order of the membrane. Interestingly, although no neuronal damage could be demonstrated, oAβ was found to trigger bifurcated phosphoinositide-dependent signaling in the neuron, on one hand, the phosphorylation of insulin receptor, the phosphatidylinositol 3-kinase (PI3K)-dependent activation of Akt, its translocation to the nucleus and the concomitant phosphorylation, inactivation, and nuclear exclusion of the transcription factor Forkhead Box O3a (FoxO3a), and on the other, phosphoinositide-phospholipase C (PI-PLC)-dependent extracellular signal-regulated kinase 1/2 (ERK1/2) activation. Pharmacological manipulation of the signaling cascades was used in order to better characterize the role of oAβ-activated signals, and mitochondrial function was determined as a measure of neuronal viability. The inhibition of PI3K, PI-PLC, and general phosphoinositide metabolism impaired neuronal mitochondrial function. Furthermore, increased oAβ-induced cell death was observed in the presence of phosphoinositide metabolism inhibition. Our results allow us to conclude that oAβ triggers the activation of phosphoinositide-dependent signaling, which results in the subsequent activation of neuroprotective mechanisms that could be involved in the determination of neuronal fate.
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Abbreviations
- AD:
-
Alzheimer’s disease
- ANOVA:
-
Analysis of variance
- APP:
-
Amyloid beta precursor protein
- Aβ:
-
Amyloid β peptide
- BSA:
-
Bovine serum albumin
- DAG:
-
Diacylglycerol
- DCDCDHF:
-
2′,7′-Dichlorofluorescein diacetate
- DMEM:
-
Dulbecco’s modified Eagle’s medium
- DMSO:
-
Dimethyl sulfoxide
- DPH:
-
1,6-diphenyl-1,3,5-hexatrien
- DTT:
-
Dithiothreitol
- EDTA:
-
Ethylenediaminetetraacetic acid
- EM:
-
Electron microscopy
- ERK1/2:
-
Extracellular signal-regulated kinases 1/2
- FBS:
-
Fetal bovine serum
- FoxO3a:
-
Forkhead Box O3a
- GP:
-
Generalized polarization
- hNRP:
-
Human NAP-related protein
- HRP:
-
Horseradish peroxidase
- IR:
-
Insulin receptor
- LDH:
-
Lactate dehydrogenase
- LilrB2:
-
Leukocyte immunoglobulin-like receptor B2
- MAPK:
-
Mitogen-activated protein kinase
- MTT:
-
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
- oAβ:
-
Oligomeric amyloid β peptide
- PBS:
-
Phosphate-buffered saline
- PI:
-
Phosphoinositide
- PI3K:
-
Phosphatidylinositol 3-kinase
- PIP2 :
-
Phosphatidylinositol bis-phosphate
- PI-PLC:
-
Phosphoinositide-phospholipase C
- PrPC :
-
Cellular prion protein
- TBA:
-
Thiobarbituric acid
- TBARS:
-
Thiobarbituric acid-reactive substance
- TCA:
-
Trichloroacetic acid
- TMA-DPH:
-
1-(4-trimethylammoniumphenyl)-6-phenyl-1,3,5-hexatriene
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Acknowledgments
We thank Dr. Charles Glabe (Department of Molecular Biology and Biochemistry, University of California, Irvine, USA) for kindly providing OC and A11 antibodies. This work was supported by Universidad Nacional del Sur (PGI 24/B179), Agencia Nacional de Promoción Científica y Tecnológica (ANPCyT; PICT-2010-0936 and PICT-2013-0987), and Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET; PIP 11220120100251).
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Natalia Paola Alza and Melisa Ailén Conde contributed equally to this work.
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Supplementary Fig. 1
Negative control, no effect on PI3K was exerted by non-oligomerized Aβ (TIF 1120 kb)
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Uranga, R.M., Alza, N.P., Conde, M.A. et al. Phosphoinositides: Two-Path Signaling in Neuronal Response to Oligomeric Amyloid β Peptide. Mol Neurobiol 54, 3236–3252 (2017). https://doi.org/10.1007/s12035-016-9885-3
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DOI: https://doi.org/10.1007/s12035-016-9885-3