Macroautophagy in sporadic and the genetic form of Parkinson's disease with the A53T α-synuclein mutation

Translational Neurodegeneration

Glenda M Halliday²  Weiping Gai¹  Karen Murphy²  Germaine Chua²  Fariba Chegini¹  Yue Huang² 

[1] Department of Human Physiology and the Centre for Neuroscience, Flinders University School of Medicine, Bedford Park, SA5042, Australia;Neuroscience Research Australia and the University of New South Wales, Sydney, 2031, Australia

关键词: Parkinson's disease;    macroautophagy;    α-synuclein;   

Others  :  839058
DOI  :  10.1186/2047-9158-1-2

 received in 2011-12-06, accepted in 2012-01-13,  发布年份 2012

【 摘 要 】

Background

The A53T mutation in the α-synuclein gene causes autosomal-dominant Lewy body Parkinson's disease (PD). Cultured cell models have linked this mutation to increased cell macroautophagy, although evidence of enhanced macroautophagy in patients with this mutation has not been assessed.

Objective

To determine whether macroautophagy is increased by the A53T α-synuclein gene mutation in PD patients and cell models.

Methods

Formalin-fixed paraffin-embedded 10 μm-thick tissue sections from the substantia nigra and anterior cingulate cortex of two PD patients with the A53T α-synuclein gene mutation were compared with four sporadic PD cases and four controls obtained from the Sydney Brain Bank. Lewy bodies were isolated from frontal cortex of a case with late stage PD (recruited from South Australian Brain Bank). Immunohistochemistry was performed for α-synuclein and the macroautophagy markers autophagy-specific gene (ATG) 5, ATG6/Beclin1 and ATG8/LC3. SH-SY5Y cells were transfected with wild type or A53T mutant α-synuclein plasmids and observable changes in macroautophagy marker protein levels assessed using Western blotting.

Results

α-Synuclein immunoreactive neurites and dots were more numerous in patients with A53T mutations compared with late stage sporadic PD patients, and perinuclear cytoplasmic α-synuclein aggregates were observed in the α-synuclein A53T gene transfected SH-SY5Y cells compared to wild type transfections. All PD patients (with or without A53T mutations) had increased immunohistochemical evidence for macroautophagy compared with controls, and the levels of the ATG5 complex were equally increased in wild type and A53T α-synuclein gene transfected cells compared to controls.

Conclusion

Despite increased α-synuclein accumulation with A53T mutations, macroautophagy is not increased above that observed in sporadic patients with PD or in cells transfected with wild type α-synuclein, suggesting that mutated α-synuclein protein is not removed by macroautophagy.

【 授权许可】

   
2012 Huang et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

【 参考文献 】

• [1]Polymeropoulos MH, et al.: Mutation in the alpha-synuclein gene identified in families with Parkinson's disease. Science 1997, 276(5321):2045-7.
• [2]Kruger R, et al.: Ala30Pro mutation in the gene encoding alpha-synuclein in Parkinson's disease. Nat Genet 1998, 18(2):106-8.
• [3]Zarranz JJ, et al.: The new mutation, E46K, of alpha-synuclein causes Parkinson and Lewy body dementia. Ann Neurol 2004, 55(2):164-73.
• [4]Singleton AB, et al.: alpha-Synuclein locus triplication causes Parkinson's disease. Science 2003, 302(5646):841.
• [5]Huang Y, et al.: Genetic contributions to Parkinson's disease. Brain Res Brain Res Rev 2004, 46(1):44-70.
• [6]Shin CW, et al.: Two Parkinson's disease patients with alpha-synuclein gene duplication and rapid cognitive decline. Mov Disord 2010, 25(7):957-9.
• [7]Braak H, et al.: Staging of brain pathology related to sporadic Parkinson's disease. Neurobiol Aging 2003, 24(2):197-211.
• [8]Halliday G, et al.: The progression of pathology in longitudinally followed patients with Parkinson's disease. Acta Neuropathol 2008, 115(4):409-15.
• [9]McLean PJ, Kawamata H, Hyman BT: Alpha-synuclein-enhanced green fluorescent protein fusion proteins form proteasome sensitive inclusions in primary neurons. Neuroscience 2001, 104(3):901-12.
• [10]Meredith GE, et al.: Lysosomal malfunction accompanies alpha-synuclein aggregation in a progressive mouse model of Parkinson's disease. Brain Res 2002, 956(1):156-65.
• [11]Cuervo AM, et al.: Impaired degradation of mutant alpha-synuclein by chaperone mediated autophagy. Science 2004, 305(5688):1292-5.
• [12]Wilson CA, et al.: Degradative organelles containing mislocalized alpha-and beta synuclein proliferate in presenilin-1 null neurons. J Cell Biol 2004, 165(3):335-46.
• [13]Maiuri MC, Criollo A, Kroemer G: Crosstalk between apoptosis and autophagy within the Beclin 1 interactome. EMBO J 2010, 29(3):515-6.
• [14]Ma JF, et al.: Immunohistochemical evidence for macroautophagy in neurones and endothelial cells in Alzheimer's disease. Neuropathol Appl Neurobiol 2010, 36(4):312-9.
• [15]Crews L, et al.: Selective molecular alterations in the autophagy pathway in patients with Lewy body disease and in models of alpha-synucleinopathy. PLoS One 2010, 5(2):e9313.
• [16]Choubey V, et al.: Mutant A53T alpha-synuclein induces neuronal death by increasing mitochondrial autophagy. J Biol Chem 2011, 286(12):10814-24.
• [17]Chew KC, et al.: Enhanced Autophagy from Chronic Toxicity of Iron and Mutant A53T {alpha}-Synuclein: IMPLICATIONS FOR NEURONAL CELL DEATH IN PARKINSON DISEASE. J Biol Chem 2011, 286(38):33380-9.
• [18]Gai WP, et al.: In situ and in vitro study of colocalization and segregation of alpha synuclein, ubiquitin, and lipids in Lewy bodies. Exp Neurol 2000, 166(2):324-33.
• [19]Spira PJ, et al.: Clinical and pathological features of a Parkinsonian syndrome in a family with an Ala53Thr alpha-synuclein mutation. Ann Neurol 2001, 49(3):313-9.
• [20]Kabeya Y, et al.: LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. EMBO J 2000, 19(21):5720-8.
• [21]Russo R, et al.: Calpain-mediated cleavage of Beclin-1 and autophagy deregulation following retinal ischemic injury in vivo. Cell Death Dis 2011, 2:e144.
• [22]Markopoulou K, et al.: Clinical, neuropathological and genotypic variability in SNCA A53T familial Parkinson's disease. Variability in familial Parkinson's disease. Acta Neuropathol 2008, 116(1):25-35.
• [23]Giasson BI, et al.: Neuronal alpha-synucleinopathy with severe movement disorder in mice expressing A53T human alpha-synuclein. Neuron 2002, 34(4):521-33.
• [24]Stefanis L, et al.: Expression of A53T mutant but not wild-type alpha-synuclein in PC12 cells induces alterations of the ubiquitin-dependent degradation system, loss of dopamine release, and autophagic cell death. J Neurosci 2001, 21(24):9549-60.
• [25]Conway KA, Harper JD, Lansbury PT: Accelerated in vitro fibril formation by a mutant alpha-synuclein linked to early-onset Parkinson disease. Nat Med 1998, 4(11):1318-20.
• [26]Webb JL, et al.: Alpha-Synuclein is degraded by both autophagy and the proteasome. J Biol Chem 2003, 278(27):25009-13.
• [27]Vogiatzi T, et al.: Wild type alpha-synuclein is degraded by chaperone-mediated autophagy and macroautophagy in neuronal cells. J Biol Chem 2008, 283(35):23542-56.
• [28]Yang F, et al.: Role of autophagy and proteasome degradation pathways in apoptosis of PC12 cells overexpressing human alpha-synuclein. Neurosci Lett 2009, 454(3):203-8.
• [29]Prigione A, et al.: Alpha-synuclein nitration and autophagy response are induced in peripheral blood cells from patients with Parkinson disease. Neurosci Lett 2010, 477(1):6-10.
• [30]Su M, et al.: HDAC6 regulates aggresome-autophagy degradation pathway of alpha synuclein in response to MPP+-induced stress. J Neurochem 2011, 117(1):112-20.
• [31]Xilouri M, et al.: Abberant alpha-synuclein confers toxicity to neurons in part through inhibition of chaperone-mediated autophagy. PLoS One 2009, 4(5):e5515.
• [32]Martinez-Vicente M, et al.: Dopamine-modified alpha-synuclein blocks chaperone mediated autophagy. J Clin Invest 2008, 118(2):777-88.
• [33]Sarkar S, et al.: Trehalose, a novel mTOR-independent autophagy enhancer, accelerates the clearance of mutant huntingtin and alpha-synuclein. J Biol Chem 2007, 282(8):5641-52.
• [34]Winslow AR, et al.: alpha-Synuclein impairs macroautophagy: implications for Parkinson's disease. J Cell Biol 2010, 190(6):1023-37.
• [35]Tanji K, et al.: Alteration of autophagosomal proteins (LC3, GABARAP and GATE-16) in Lewy body disease. Neurobiol Dis 2011, 43(3):690-7.
• [36]Van Humbeeck C, et al.: Parkin interacts with Ambra1 to induce mitophagy. J Neurosci 2011, 31(28):10249-61.
• [37]Shi G, et al.: Functional alteration of PARL contributes to mitochondrial dysregulation in Parkinson's disease. Hum Mol Genet 2011, 20(10):1966-74.
• [38]Loeb V, et al.: The transgenic overexpression of alpha-synuclein and not its related pathology associates with complex I inhibition. J Biol Chem 2010, 285(10):7334-43.
• [39]Esteves AR, et al.: Dysfunctional mitochondria uphold calpain activation: contribution to Parkinson's disease pathology. Neurobiol Dis 2010, 37(3):723-30.