Top

Published in:

Open Access 01-03-2012 | Basic Neurosciences, Genetics and Immunology - Original Article

A53T-alpha-synuclein-overexpression in the mouse nigrostriatal pathway leads to early increase of 14-3-3 epsilon and late increase of GFAP

Authors: Alexander Kurz, Caroline May, Oliver Schmidt, Thorsten Müller, Christian Stephan, Helmut E. Meyer, Suzana Gispert, Georg Auburger, Katrin Marcus

Published in: Journal of Neural Transmission | Issue 3/2012

Abstract

Parkinson’s disease (PD) is a neurodegenerative disorder frequent at old age characterized by atrophy of the nigrostriatal projection. Overexpression and A53T-mutation of the presynaptic, vesicle-associated chaperone alpha-synuclein are known to cause early-onset autosomal dominant PD. We previously generated mice with transgenic overexpression of human A53T-alpha-synuclein (A53T-SNCA) in dopaminergic substantia nigra neurons as a model of early PD. To elucidate the early and late effects of A53T-alpha-synuclein on the proteome of dopaminergic nerve terminals in the striatum, we now investigated expression profiles of young and old mice using two-dimensional fluorescence difference in gel electrophoresis (2D-DIGE) and mass spectrometry. In total, 15 proteins were upregulated and 2 downregulated. Mice before the onset of motor anomalies showed an upregulation of the spot containing 14-3-3 proteins, in particular the epsilon isoform, as well as altered levels of chaperones, vesicle trafficking and bioenergetics proteins. In old mice, the persistent upregulation of 14-3-3 proteins was aggravated by an increase of glial fibrillary acidic protein (GFAP) suggesting astrogliosis due to initial neurodegeneration. Independent immunoblots corroborated GFAP upregulation and 14-3-3 upregulation for the epsilon isoform, and also detected significant eta and gamma changes. Only for 14-3-3 epsilon a corresponding mRNA increase was observed in midbrain, suggesting it is transcribed in dopaminergic perikarya and accumulates as protein in presynapses, together with A53T-SNCA. 14-3-3 proteins associate with alpha-synuclein in vitro and in pathognomonic Lewy bodies of PD brains. They act as chaperones in signaling, dopamine synthesis and stress response. Thus, their early dysregulation probably reflects a response to alpha-synuclein toxicity.

Available only for authorised users

Ahmed N, Battah S, Karachalias N, Babaei-Jadidi R, Horanyi M, Baroti K, Hollan S, Thornalley PJ (2003) Increased formation of methylglyoxal and protein glycation, oxidation and nitrosation in triosephosphate isomerase deficiency. Biochim Biophys Acta 1639:121–132PubMed

Baumann S, Ceglarek U, Fiedler GM, Lembcke J, Leichtle A, Thiery J (2005) Standardized approach to proteome profiling of human serum based on magnetic bead separation and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Clin Chem 51:973–980PubMedCrossRef

Berg D, Holzmann C, Riess O (2003) 14–3-3 proteins in the nervous system. Nat Rev Neurosci 4:752–762PubMedCrossRef

Braak H, Del Tredici K, Rub U, de Vos RA, Jansen Steur EN, Braak E (2003) Staging of brain pathology related to sporadic Parkinson’s disease. Neurobiol Aging 24:197–211PubMedCrossRef

Bussell R Jr, Eliezer D (2003) A structural and functional role for 11-mer repeats in alpha-synuclein and other exchangeable lipid binding proteins. J Mol Biol 329:763–778PubMedCrossRef

Butor C, Diaz S, Varki A (1993) High level O-acetylation of sialic acids on N-linked oligosaccharides of rat liver membranes. Differential subcellular distribution of 7- and 9-O-acetyl groups and of enzymes involved in their regulation. J Biol Chem 268:10197–10206PubMed

Cabin DE, Gispert-Sanchez S, Murphy D, Auburger G, Myers RR, Nussbaum RL (2005) Exacerbated synucleinopathy in mice expressing A53T SNCA on a Snca null background. Neurobiol Aging 26:25–35PubMedCrossRef

Chamrad DC, Koerting G, Gobom J, Thiele H, Klose J, Meyer HE, Blueggel M (2003) Interpretation of mass spectrometry data for high-throughput proteomics. Anal Bioanal Chem 376:1014–1022PubMedCrossRef

Chandra S, Fornai F, Kwon HB, Yazdani U, Atasoy D, Liu X, Hammer RE, Battaglia G, German DC, Castillo PE, Sudhof TC (2004) Double-knockout mice for alpha- and beta-synucleins: effect on synaptic functions. Proc Natl Acad Sci USA 101:14966–14971PubMedCrossRef

Chandra S, Gallardo G, Fernandez-Chacon R, Schluter OM, Sudhof TC (2005) Alpha-synuclein cooperates with CSPalpha in preventing neurodegeneration. Cell 123:383–396PubMedCrossRef

Clayton DF, George JM (1998) The synucleins: a family of proteins involved in synaptic function, plasticity, neurodegeneration and disease. Trends Neurosci 21:249–254PubMedCrossRef

Conway KA, Harper JD, Lansbury PT (1998) Accelerated in vitro fibril formation by a mutant alpha-synuclein linked to early-onset Parkinson disease. Nat Med 4:1318–1320PubMedCrossRef

Crowther DC (2002) Familial conformational diseases and dementias. Hum Mutat 20:1–14PubMedCrossRef

Dzamko N, Deak M, Hentati F, Reith AD, Prescott AR, Alessi DR, Nichols RJ (2010) Inhibition of LRRK2 kinase activity leads to dephosphorylation of Ser(910)/Ser(935), disruption of 14–3-3 binding and altered cytoplasmic localization. Biochem J 430:405–413PubMedCrossRef

Eng JM, McCornack AL, Yates JR 3rd (1994) An approach to correlate tandem mass-spectral data of peptides with amino-acid-sequences in a protein database. J Am Soc Mass Spectrom 5:976–989CrossRef

Fahn S (2003) Description of Parkinson’s disease as a clinical syndrome. Ann N Y Acad Sci 991:1–14PubMedCrossRef

Ferguson SM, Brasnjo G, Hayashi M, Wolfel M, Collesi C, Giovedi S, Raimondi A, Gong LW, Ariel P, Paradise S, O’Toole E, Flavell R, Cremona O, Miesenbock G, Ryan TA, De Camilli P (2007) A selective activity-dependent requirement for dynamin 1 in synaptic vesicle endocytosis. Science 316:570–574PubMedCrossRef

Garcia-Reitbock P, Anichtchik O, Bellucci A, Iovino M, Ballini C, Fineberg E, Ghetti B, Della Corte L, Spano P, Tofaris GK, Goedert M, Spillantini MG (2010) SNARE protein redistribution and synaptic failure in a transgenic mouse model of Parkinson’s disease. Brain 133:2032–2044PubMedCrossRef

Gasser T (2001) Genetics of Parkinson’s disease. J Neurol 248:833–840PubMedCrossRef

Gillardon F, Mack M, Rist W, Schnack C, Lenter M, Hildebrandt T, Hengerer B (2008) MicroRNA and proteome expression profiling in early-symptomatic alpha-synuclein (A30P)-transgenic mice. Proteomics 2:697–705PubMedCrossRef

Gispert S, Del Turco D, Garrett L, Chen A, Bernard DJ, Hamm-Clement J, Korf HW, Deller T, Braak H, Auburger G, Nussbaum RL (2003) Transgenic mice expressing mutant A53T human alpha-synuclein show neuronal dysfunction in the absence of aggregate formation. Mol Cell Neurosci 24:419–429PubMedCrossRef

Huang Y, Cheung L, Rowe D, Halliday G (2004) Genetic contributions to Parkinson’s disease. Brain Res Brain Res Rev 46:44–70PubMedCrossRef

Ichibangase T, Saimaru H, Takamura N, Kuwahara T, Koyama A, Iwatsubo T, Imai K (2008) Proteomics of Caenorhabditis elegans over-expressing human alpha-synuclein analyzed by fluorogenic derivatization-liquid chromatography/tandem mass spectrometry: identification of actin and several ribosomal proteins as negative markers at early Parkinson‘s disease stages. Biomed Chromatogr 22:232–234PubMedCrossRef

Kahle PJ, Neumann M, Ozmen L, Muller V, Odoy S, Okamoto N, Jacobsen H, Iwatsubo T, Trojanowski JQ, Takahashi H, Wakabayashi K, Bogdanovic N, Riederer P, Kretzschmar HA, Haass C (2001) Selective insolubility of alpha-synuclein in human Lewy body diseases is recapitulated in a transgenic mouse model. Am J Pathol 159:2215–2225PubMedCrossRef

Karp NA, McCormick PS, Russell MR, Lilley KS (2007) Experimental and statistical considerations to avoid false conclusions in proteomics studies using differential in-gel electrophoresis. Mol Cell Proteom 6:1354–1364CrossRef

Kessler JC, Rochet JC, Lansbury PT Jr (2003) The N-terminal repeat domain of alpha-synuclein inhibits beta-sheet and amyloid fibril formation. Biochemistry 42:672–678PubMedCrossRef

Klose J, Kobalz U (1995) Two-dimensional electrophoresis of proteins: an updated protocol and implications for a functional analysis of the genome. Electrophoresis 16:1034–1059PubMedCrossRef

Kurz A, Double KL, Lastres-Becker I, Tozzi A, Tantucci M, Bockhart V, Bonin M, García-Arencibia M, Nuber S, Schlaudraff F, Liss B, Fernández-Ruiz J, Gerlach M, Wüllner U, Lüddens H, Calabresi P, Auburger G, Gispert S (2010) A53T-alpha-synuclein overexpression impairs dopamine signaling and striatal synaptic plasticity in old mice. PLoS ONE 5:e11464PubMedCrossRef

Leroy E, Boyer R, Auburger G, Leube B, Ulm G, Mezey E, Harta G, Brownstein MJ, Jonnalagada S, Chernova T, Dehejia A, Lavedan C, Gasser T, Steinbach PJ, Wilkinson KD, Polymeropoulos MH (1998) The ubiquitin pathway in Parkinson’s disease. Nature 395:451–452PubMedCrossRef

Licker V, Kovari E, Hochstrasser DF, Burkhard PR (2009) Proteomics in human Parkinson’s disease research. J Proteom 73:10–29CrossRef

Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25:402–408PubMedCrossRef

Maries E, Dass B, Collier TJ, Kordower JH, Steece-Collier K (2003) The role of alpha-synuclein in Parkinson’s disease: insights from animal models. Nat Rev Neurosci 4:727–738PubMedCrossRef

Martin H, Rostas J, Patel Y, Aitken A (1994) Subcellular localisation of 14-3-3 isoforms in rat brain using specific antibodies. J Neurochem 63:2259–2265PubMedCrossRef

Narhi L, Wood SJ, Steavenson S, Jiang Y, Wu GM, Anafi D, Kaufman SA, Martin F, Sitney K, Denis P, Louis JC, Wypych J, Biere AL, Citron M (1999) Both familial Parkinson’s disease mutations accelerate alpha-synuclein aggregation. J Biol Chem 274:9843–9846PubMedCrossRef

Nichols RJ, Dzamko N, Morrice NA, Campbell DG, Deak M, Ordureau A, Macartney T, Tong Y, Shen J, Prescott AR, Alessi DR (2010) 14–3-3 binding to LRRK2 is disrupted by multiple Parkinson‘s disease-associated mutations and regulates cytoplasmic localization. Biochem J 430:393–404PubMedCrossRef

Olanow CW, McNaught KS (2006) Ubiquitin-proteasome system and Parkinson’s disease. Mov Disord 21:1806–1823PubMedCrossRef

Ostrerova N, Petrucelli L, Farrer M, Mehta N, Choi P, Hardy J, Wolozin B (1999) alpha-Synuclein shares physical and functional homology with 14-3-3 proteins. J Neurosci 19:5782–5791PubMed

Palacino JJ, Sagi D, Goldberg MS, Krauss S, Motz C, Wacker M, Klose J, Shen J (2004) Mitochondrial dysfunction and oxidative damage in parkin-deficient mice. J Biol Chem 279:18614–18622PubMedCrossRef

Paxinos G, Franklin KBJ (2001) The Mouse Brain in Stereotaxic Coordinates., Second edn. Academic Press, San Diego

Pennington K, Peng J, Hung CC, Banks RE, Robinson PA (2010) Differential effects of wild-type and A53T mutant isoform of alpha-synuclein on the mitochondrial proteome of differentiated SH-SY5Y cells. J Proteom Res 9:2390–2401CrossRef

Perez RG, Waymire JC, Lin E, Liu JJ, Guo F, Zigmond MJ (2002) A role for alpha-synuclein in the regulation of dopamine biosynthesis. J Neurosci 22:3090–3099PubMed

Perkins DN, Pappin DJ, Creasy DM, Cottrell JS (1999) Probability-based protein identification by searching sequence databases using mass spectrometry data. Electrophoresis 20:3551–3567PubMedCrossRef

Poll-The BT, Aicardi J, Girot R, Rosa R (1985) Neurological findings in triosephosphate isomerase deficiency. Ann Neurol 17:439–443PubMedCrossRef

Polymeropoulos MH, Lavedan C, Leroy E, Ide SE, Dehejia A, Dutra A, Pike B, Root H, Rubenstein J, Boyer R, Stenroos ES, Chandrasekharappa S, Athanassiadou A, Papapetropoulos T, Johnson WG, Lazzarini AM, Duvoisin RC, Di Iorio G, Golbe LI, Nussbaum RL (1997) Mutation in the alpha-synuclein gene identified in families with Parkinson’s disease. Science 276:2045–2047PubMedCrossRef

Poon HF, Frasier M, Shreve N, Calabrese V, Wolozin B, Butterfield DA (2005) Mitochondrial associated metabolic proteins are selectively oxidized in A30P alpha-synuclein transgenic mice–a model of familial Parkinson’s disease. Neurobiol Dis 18:492–498PubMedCrossRef

Reynolds AD, Stone DK, Mosley RL, Gendelman HE (2009) Proteomic studies of nitrated alpha-synuclein microglia regulation by CD4+ CD25+ T cells. J Proteome Res 8:3497–3511PubMedCrossRef

Rudenko IN, Cookson MR (2010) 14–3-3 proteins are promising LRRK2 interactors. Biochem J 430:e5–e6PubMedCrossRef

Schaefer H, Marcus K, Sickmann A, Herrmann M, Klose J, Meyer HE (2003) Identification of phosphorylation and acetylation sites in alphaA-crystallin of the eye lens (mus musculus) after two-dimensional gel electrophoresis. Anal Bioanal Chem 376:966–972PubMedCrossRef

Schaefer H, Chervet JP, Bunse C, Joppich C, Meyer HE, Marcus K (2004) A peptide preconcentration approach for nano-high-performance liquid chromatography to diminish memory effects. Proteomics 4:2541–2544PubMedCrossRef

Schaefer H, Chamrad DC, Herrmann M, Stuwe J, Becker G, Klose J, Blueggel M, Meyer HE, Marcus K (2006) Study of posttranslational modifications in lenticular alphaA-Crystallin of mice using proteomic analysis techniques. Biochim Biophys Acta 1764:1948–1962PubMed

Schapira AH, Tolosa E (2010) Molecular and clinical prodrome of Parkinson disease: implications for treatment. Nat Rev Neurol 6:309–317PubMedCrossRef

Setsuie R, Wada K (2007) The functions of UCH-L1 and its relation to neurodegenerative diseases. Neurochem Int 51:105–111PubMedCrossRef

Shirakashi Y, Kawamoto Y, Tomimoto H, Takahashi R, Ihara M (2006) alpha-Synuclein is colocalized with 14-3-3 and synphilin-1 in A53T transgenic mice. Acta Neuropathol 112:681–689PubMedCrossRef

Singleton AB, Farrer M, Johnson J, Singleton A, Hague S, Kachergus J, Hulihan M, Peuralinna T, Dutra A, Nussbaum R, Lincoln S, Crawley A, Hanson M, Maraganore D, Adler C, Cookson MR, Muenter M, Baptista M, Miller D, Blancato J, Hardy J, Gwinn-Hardy K (2003) Alpha-synuclein locus triplication causes Parkinson’s disease. Science 302:841PubMedCrossRef

Sitek B, Apostolov O, Stuhler K, Pfeiffer K, Meyer HE, Eggert A, Schramm A (2005) Identification of dynamic proteome changes upon ligand activation of Trk-receptors using two-dimensional fluorescence difference gel electrophoresis and mass spectrometry. Mol Cell Proteom 4:291–299CrossRef

Spillantini MG, Schmidt ML, Lee VM, Trojanowski JQ, Jakes R, Goedert M (1997) Alpha-synuclein in Lewy bodies. Nature 388:839–840PubMedCrossRef

Stephan C, Reidegeld KA, Hamacher M, van Hall A, Marcus K, Taylor C, Jones P, Muller M, Apweiler R, Martens L, Korting G, Chamrad DC, Thiele H, Bluggel M, Parkinson D, Binz PA, Lyall A, Meyer HE (2006) Automated reprocessing pipeline for searching heterogeneous mass spectrometric data of the HUPO Brain Proteome Project pilot phase. Proteomics 6:5015–5029PubMedCrossRef

Stuhler K, Pfeiffer K, Joppich C, Stephan C, Jung K, Muller M, Schmidt O, van Hall A, Hamacher M, Urfer W, Meyer HE, Marcus K (2006) Pilot study of the Human Proteome Organisation Brain Proteome Project: applying different 2-DE techniques to monitor proteomic changes during murine brain development. Proteomics 6:4899–4913PubMedCrossRef

Titz B, Low T, Komisopoulou E, Chen SS, Rubbi L, Graeber TG (2010) The proximal signaling network of the BCR-ABL1 oncogene shows a modular organization. Oncogene 29:5895–5910PubMedCrossRef

Volles MJ, Lansbury PT Jr (2003) Zeroing in on the pathogenic form of alpha-synuclein and its mechanism of neurotoxicity in Parkinson’s disease. Biochemistry 42:7871–7878PubMedCrossRef

Wiesinger H, Hamprecht B, Dringen R (1997) Metabolic pathways for glucose in astrocytes. Glia 21:22–34PubMedCrossRef

Xu J, Kao SY, Lee FJ, Song W, Jin LW, Yankner BA (2002) Dopamine-dependent neurotoxicity of alpha-synuclein: a mechanism for selective neurodegeneration in Parkinson disease. Nat Med 8:600–606PubMedCrossRef

Xun Z, Sowell RA, Kaufman TC, Clemmer DE (2007a) Lifetime proteomic profiling of an A30P alpha-synuclein Drosophila model of Parkinson’s disease. J Proteome Res 6:3729–3738PubMedCrossRef

Xun Z, Sowell RA, Kaufman TC, Clemmer DE (2007b) Protein expression in a Drosophila model of Parkinson’s disease. J Proteome Res 6:348–357PubMedCrossRef

Xun Z, Sowell RA, Kaufman TC, Clemmer DE (2008) Quantitative proteomics of a presymptomatic A53T alpha-synuclein Drosophila model of Parkinson disease. Mol Cell Proteom 7:1191–1203CrossRef

Yacoubian TA, Slone SR, Harrington AJ, Hamamichi S, Schieltz JM, Caldwell KA, Caldwell GA, Standaert DG (2011) Differential neuroprotective effects of 14-3-3 proteins in models of Parkinson’s disease. Cell Death Dis 1

Yates JR 3rd, Eng JK, McCormack AL, Schieltz D (1995) Method to correlate tandem mass spectra of modified peptides to amino acid sequences in the protein database. Anal Chem 67:1426–1436PubMedCrossRef

Zhang W, Chait BT (2000) ProFound: an expert system for protein identification using mass spectrometric peptide mapping information. Anal Chem 72:2482–2489PubMedCrossRef

Zhou Y, Gu G, Goodlett DR, Zhang T, Pan C, Montine TJ, Montine KS, Aebersold RH, Zhang J (2004) Analysis of alpha-synuclein-associated proteins by quantitative proteomics. J Biol Chem 279:39155–39164PubMedCrossRef

Title: A53T-alpha-synuclein-overexpression in the mouse nigrostriatal pathway leads to early increase of 14-3-3 epsilon and late increase of GFAP
Authors: Alexander Kurz
Caroline May
Oliver Schmidt
Thorsten Müller
Christian Stephan
Helmut E. Meyer
Suzana Gispert
Georg Auburger
Katrin Marcus
Publication date: 01-03-2012
Publisher: Springer Vienna
Published in: Journal of Neural Transmission / Issue 3/2012
Print ISSN: 0300-9564
Electronic ISSN: 1435-1463
DOI: https://doi.org/10.1007/s00702-011-0717-3

At a glance: The STEP trials

Springer Medicine

A53T-alpha-synuclein-overexpression in the mouse nigrostriatal pathway leads to early increase of 14-3-3 epsilon and late increase of GFAP

Abstract

At a glance: The STEP trials

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 3/2012

Rescue of social behavior impairment by clozapine and alterations in the expression of neuronal receptors in a rat model of neurodevelopmental impairment induced by GRPR blockade

The effect of intrathecal administration of glial activation inhibitors on dorsal horn BDNF overexpression and hind paw mechanical allodynia in spinal nerve ligated rats

Brain tissue echogenicity—implications for substantia nigra studies in parkinsonian patients

Administration of a selective glucocorticoid antagonist attenuates electroconvulsive shock-induced retrograde amnesia

A 14 bp indel variation in the NCX1 gene modulates the age at onset in late-onset Alzheimer’s disease

ApoE gene polymorphism and vascular dementia in Chinese population: a meta-analysis