Abstract
Alzheimer’s disease (AD), the most common cause of dementia, is neuropathologically characterized by accumulation of insoluble fibrous inclusions in the brain in the form of intracellular neurofibrillary tangles and extracellular senile plaques. Perturbation of the ubiquitin-proteasome system (UPS) has long been considered an attractive hypothesis to explain the pathogenesis of AD. However, studies on UPS functionality with various methods and AD models have achieved non-conclusive results. To get further insight into UPS functionality in AD, we have crossed a well-documented APPswe/PS1dE9 AD mouse model with a UPS functionality reporter, GFPu, mouse expressing green fluorescence protein (GFP) fused to a constitutive degradation signal (CL-1) that facilitates its rapid turnover in conditions of a normal UPS. Our western blot results indicate that GFPu reporter protein was accumulated in the cortex and hippocampus, but not striatum in the APPswe/PS1dE9 AD mouse model at 4 weeks of age, which is confirmed by fluorescence microscopy and elevated levels of p53, an endogenous UPS substrate. In accordance with this, the levels of ubiquitinated proteins were elevated in the AD mouse model. These results suggest that UPS is either impaired or functionally insufficient in specific brain regions in the APPswe/PS1dE9 AD mouse model at a very young age, long before senile plaque formation and the onset of memory loss. These observations may shed new light on the pathogenesis of AD.
Abbreviations
- AD:
-
Alzheimer’s disease
- UPS:
-
Ubiquitin-proteasome system
- GFPu:
-
Green fluorescence reporter for UPS functionality
- APP:
-
Amyloid precursor protein
- PS:
-
Presenilin
- NFT:
-
Neurofibrillary tangle
- PolyUb:
-
Polyubiquitin
- Tg:
-
Transgenic
References
Baek BS, Kwon HJ, Lee KH, Yoo MA, Kim KW, Ikeno Y, Yu BP, Chung HY (1999) Regional difference of ROS generation, lipid peroxidation, and antioxidant enzyme activity in rat brain and their dietary modulation. Arch Pharmacal Res 22(4):361–366
Bence NF, Sampat RM, Kopito RR (2001) Impairment of the ubiquitin-proteasome system by protein aggregation. Science 292(5521):1552–1555. doi:10.1126/science.292 5521.1552
Bence NF, Bennett EJ, Kopito RR (2005) Application and analysis of the GFPu family of ubiquitin-proteasome system reporters. Methods Enzymol 399:481–490. doi:10.1016/S0076-6879(05)99033-2
Bennett EJ, Bence NF, Jayakumar R, Kopito RR (2005) Global impairment of the ubiquitin-proteasome system by nuclear or cytoplasmic protein aggregates precedes inclusion body formation. Mol Cell 17(3):351–365. doi:10.1016/j.molcel.2004.12.021
Bett JS, Cook C, Petrucelli L, Bates GP (2009) The ubiquitin-proteasome reporter GFPu does not accumulate in neurons of the R6/2 transgenic mouse model of Huntington’s disease. PLoS ONE 4(4):e5128. doi:10.1371/journal.pone.0005128
Bird TD (2008) Genetic aspects of Alzheimer disease. Genet Med 10(4):231–239. doi:10.1097/GIM.0b013e31816b64dc
Braak H, Braak E (1991) Neuropathological stageing of Alzheimer-related changes. Acta Neuropathol 82(4):239–259
Chen Y, Neve RL, Liu H (2012) Neddylation dysfunction in Alzheimer’s disease. J Cell Mol Med 16(11):2583–2591. doi:10.1111/j.1582-4934.2012.01604.x
Crivello NA, Rosenberg IH, Shukitt-Hale B, Bielinski D, Dallal GE, Joseph JA (2007) Aging modifies brain region-specific vulnerability to experimental oxidative stress induced by low dose hydrogen peroxide. Age Dordr 29(4):191–203. doi:10.1007/s11357-007-9039-7
Dong G, Callegari EA, Gloeckner CJ, Ueffing M, Wang H (2012) Prothymosin-alpha interacts with mutant huntingtin and suppresses its cytotoxicity in cell culture. J Biol Chem 287(2):1279–1289. doi:10.1074/jbc.M111.294280
Garcia-Alloza M, Robbins EM, Zhang-Nunes SX, Purcell SM, Betensky RA, Raju S, Prada C, Greenberg SM, Bacskai BJ, Frosch MP (2006) Characterization of amyloid deposition in the APPswe/PS1dE9 mouse model of Alzheimer disease. Neurobiol Dis 24(3):516–524. doi:10.1016/j.nbd.2006.08.017
Hensley K, Hall N, Subramaniam R, Cole P, Harris M, Aksenov M, Aksenova M, Gabbita SP, Wu JF, Carney JM et al (1995) Brain regional correspondence between Alzheimer’s disease histopathology and biomarkers of protein oxidation. J Neurochem 65(5):2146–2156
Ihara Y, Morishima-Kawashima M, Nixon R (2012) The ubiquitin-proteasome system and the autophagic-lysosomal system in Alzheimer disease. Cold Spring Harb Perspect Med 2(8):1741–1751. doi:10.1101/cshperspect.a006361
Jankowsky JL, Fadale DJ, Anderson J, Xu GM, Gonzales V, Jenkins NA, Copeland NG, Lee MK, Younkin LH, Wagner SL, Younkin SG, Borchelt DR (2004) Mutant presenilins specifically elevate the levels of the 42 residue beta-amyloid peptide in vivo: evidence for augmentation of a 42-specific gamma secretase. Hum Mol Genet 13(2):159–170. doi:10.1093/hmg/ddh019
Johnston JA, Ward CL, Kopito RR (1998) Aggresomes: a cellular response to misfolded proteins. J Cell Biol 143(7):1883–1898
Keller JN, Hanni KB, Markesbery WR (2000) Impaired proteasome function in Alzheimer’s disease. J Neurochem 75(1):436–439
Khan LA, Bauer PO, Miyazaki H, Lindenberg KS, Landwehrmeyer BG, Nukina N (2006) Expanded polyglutamines impair synaptic transmission and ubiquitin-proteasome system in Caenorhabditis elegans. J Neurochem 98(2):576–587. doi:10.1111/j.1471-4159.2006.03895.x
Kumarapeli AR, Horak KM, Glasford JW, Li J, Chen Q, Liu J, Zheng H, Wang X (2005) A novel transgenic mouse model reveals deregulation of the ubiquitin-proteasome system in the heart by doxorubicin. FASEB J 19(14):2051–2053
Layfield R, Lowe J, Bedford L (2005) The ubiquitin-proteasome system and neurodegenerative disorders. Essays Biochem 41:157–171. doi:10.1042/EB0410157
Lu L, Wang H (2012) Transient focal cerebral ischemia upregulates immunoproteasomal subunits. Cell Mol Neurobiol 32(6):965–970. doi:10.1007/s10571-012-9854-y
Marone M, Mozzetti S, De Ritis D, Pierelli L, Scambia G (2001) Semiquantitative RT-PCR analysis to assess the expression levels of multiple transcripts from the same sample. Biol Proced Online 3:19–25. doi:10.1251/bpo20
Mori H, Kondo J, Ihara Y (1987) Ubiquitin is a component of paired helical filaments in Alzheimer’s disease. Science 235(4796):1641–1644
Orre M, Kamphuis W, Dooves S, Kooijman L, Chan ET, Kirk CJ, Dimayuga Smith V, Koot S, Mamber C, Jansen AH, Ovaa H, Hol EM (2013) Reactive glia show increased immunoproteasome activity in Alzheimer’s disease. Brain 136(Pt 5):1415–1431. doi:10.1093/brain/awt083
Perry G, Rizzuto N, Autilio-Gambetti L, Gambetti P (1985) Paired helical filaments from Alzheimer disease patients contain cytoskeletal components. Proc Natl Acad Sci USA 82(11):3916–3920
Schubert D, Soucek T, Blouw B (2009) The induction of HIF-1 reduces astrocyte activation by amyloid beta peptide. Eur J Neurosc 29(7):1323–1334. doi:10.1111/j.1460-9568.2009.06712.x
Su H, Li J, Menon S, Liu J, Kumarapeli AR, Wei N, Wang X (2011) Perturbation of cullin deneddylation via conditional Csn8 ablation impairs the ubiquitin-proteasome system and causes cardiomyocyte necrosis and dilated cardiomyopathy in mice. Cir Res 108(1):40–50. doi:10.1161/CIRCRESAHA.110.230607
Upadhya SC, Hegde AN (2007) Role of the ubiquitin proteasome system in Alzheimer’s disease. BMC Biochem 8(1):12. doi:10.1186/1471-2091-8-S1-S12
Acknowledgments
We would like to thank Dr. Robin Miskimins for critical reading of the manuscript, Dr. Fran Day at the Imaging Core of the University of South Dakota for help in fluorescence microscopy, and Mr. Suleman said at the histopathology core for assistance in preparation of brain sections. This work was supported by Start-up Funds from the University of South Dakota (HW).
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The authors have declared no conflicts of interest.
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Liu, Y., Hettinger, C.L., Zhang, D. et al. The Proteasome Function Reporter GFPu Accumulates in Young Brains of the APPswe/PS1dE9 Alzheimer’s Disease Mouse Model. Cell Mol Neurobiol 34, 315–322 (2014). https://doi.org/10.1007/s10571-013-0022-9
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DOI: https://doi.org/10.1007/s10571-013-0022-9