Neuron loss in the 5XFAD mouse model of Alzheimer’s disease correlates with intraneuronal Aβ42 accumulation and Caspase-3 activation

Molecular Neurodegeneration

Robert Vassar¹  William A Eimer¹ 

[1] Department of Cell and Molecular Biology, Feinberg School of Medicine, Northwestern University, 303 East Chicago Avenue, Chicago, IL, 60611, USA

关键词: Apoptosis;    Neuron loss;    Caspase-3;    Amyloid-β;    Alzheimer’s disease;    5XFAD;    Intraneuronal Aβ₄₂;   

Others  :  863288
DOI  :  10.1186/1750-1326-8-2

 received in 2012-11-21, accepted in 2013-01-09,  发布年份 2013

【 摘 要 】

Background

Although the mechanism of neuron loss in Alzheimer’s disease (AD) is enigmatic, it is associated with cerebral accumulation of Aβ₄₂. The 5XFAD mouse model of amyloid deposition expresses five familial AD (FAD) mutations that are additive in driving Aβ₄₂ overproduction. 5XFAD mice exhibit intraneuronal Aβ₄₂ accumulation at 1.5 months, amyloid deposition at 2 months, and memory deficits by 4 months of age.

Results

Here, we demonstrate by unbiased stereology that statistically significant neuron loss occurs by 9 months of age in 5XFAD mice. We validated two Aβ₄₂-selective antibodies by immunostaining 5XFAD; BACE1^−/− bigenic brain sections and then used these antibodies to show that intraneuronal Aβ₄₂ and amyloid deposition develop in the same regions where neuron loss is observed in 5XFAD brain. In 5XFAD neuronal soma, intraneuronal Aβ₄₂ accumulates in puncta that co-label for Transferrin receptor and LAMP-1, indicating endosomal and lysosomal localization, respectively. In addition, in young 5XFAD brains, we observed activated Caspase-3 in the soma and proximal dendrites of intraneuronal Aβ₄₂-labeled neurons. In older 5XFAD brains, we found activated Caspase-3-positive punctate accumulations that co-localize with the neuronal marker class III β-tubulin, suggesting neuron loss by apoptosis.

Conclusions

Together, our results indicate a temporal sequence of intraneuronal Aβ₄₂ accumulation, Caspase-3 activation, and neuron loss that implies a potential apoptotic mechanism of neuron death in the 5XFAD mouse.

【 授权许可】

   
2013 Eimer and Vassar; licensee BioMed Central Ltd.

【 图 表 】

【 参考文献 】

• [1]Bertram L, Lill CM, Tanzi RE: The genetics of Alzheimer disease: back to the future. Neuron 2010, 68:270-281.
• [2]Sisodia SS: Alzheimer’s disease: perspectives for the new millennium. J Clin Invest 1999, 104:1169-1170.
• [3]Hyslop PA, Bender MH: Methods for sample preparation for direct immunoassay measurement of analytes in tissue homogenates: ELISA assay of amyloid beta-peptides. Curr Protoc Neurosci 2002, Chapter 7:Unit 7 20.
• [4]Selkoe D, Mandelkow E, Holtzman D: Deciphering Alzheimer disease. Cold Spring Harb Perspect Med 2012, 2:a011460.
• [5]Holtzman DM, Morris JC, Goate AM: Alzheimer’s disease: the challenge of the second century. Sci Transl Med 2011, 3:77sr71.
• [6]Bertram L, Tanzi RE: The genetics of Alzheimer’s disease. Prog Mol Biol Transl Sci 2012, 107:79-100.
• [7]Gouras GK, Tampellini D, Takahashi RH, Capetillo-Zarate E: Intraneuronal beta-amyloid accumulation and synapse pathology in Alzheimer’s disease. Acta Neuropathol 2010, 119:523-541.
• [8]Wirths O, Bayer TA: Intraneuronal Abeta accumulation and neurodegeneration: lessons from transgenic models. Life Sci 2012, 91:1148-1152.
• [9]Gouras GK, Willen K, Tampellini D: Critical role of intraneuronal Abeta in Alzheimer’s disease: technical challenges in studying intracellular Abeta. Life Sci 2012, 91:1153-1158.
• [10]Oakley H, Cole SL, Logan S, Maus E, Shao P, Craft J, Guillozet-Bongaarts A, Ohno M, Disterhoft J, Van Eldik L, et al.: Intraneuronal beta-amyloid aggregates, neurodegeneration, and neuron loss in transgenic mice with five familial Alzheimer’s disease mutations: potential factors in amyloid plaque formation. J Neurosci 2006, 26:10129-10140.
• [11]Casas C, Sergeant N, Itier JM, Blanchard V, Wirths O, van der Kolk N, Vingtdeux V, van de Steeg E, Ret G, Canton T, et al.: Massive CA1/2 neuronal loss with intraneuronal and N-terminal truncated Abeta42 accumulation in a novel Alzheimer transgenic model. Am J Pathol 2004, 165:1289-1300.
• [12]Mullan M, Crawford F, Axelman K, Houlden H, Lilius L, Winblad B, Lannfelt L: A pathogenic mutation for probable Alzheimer’s disease in the APP gene at the N-terminus of beta-amyloid. Nat Genet 1992, 1:345-347.
• [13]Eckman CB, Mehta ND, Crook R, Perez-tur J, Prihar G, Pfeiffer E, Graff-Radford N, Hinder P, Yager D, Zenk B, et al.: A new pathogenic mutation in the APP gene (I716V) increases the relative proportion of A beta 42(43). Hum Mol Genet 1997, 6:2087-2089.
• [14]Goate A, Chartier-Harlin MC, Mullan M, Brown J, Crawford F, Fidani L, Giuffra L, Haynes A, Irving N, James L, et al.: Segregation of a missense mutation in the amyloid precursor protein gene with familial Alzheimer’s disease. Nature 1991, 349:704-706.
• [15]Citron M, Eckman CB, Diehl TS, Corcoran C, Ostaszewski BL, Xia W, Levesque G, St George Hyslop P, Younkin SG, Selkoe DJ: Additive effects of PS1 and APP mutations on secretion of the 42-residue amyloid beta-protein. Neurobiol Dis 1998, 5:107-116.
• [16]Jawhar S, Trawicka A, Jenneckens C, Bayer TA, Wirths O: Motor deficits, neuron loss, and reduced anxiety coinciding with axonal degeneration and intraneuronal Abeta aggregation in the 5XFAD mouse model of Alzheimer’s disease. Neurobiol Aging 2012, 33:196. e129-140
• [17]Luo Y, Bolon B, Damore MA, Fitzpatrick D, Liu H, Zhang J, Yan Q, Vassar R, Citron M: BACE1 (beta-secretase) knockout mice do not acquire compensatory gene expression changes or develop neural lesions over time. Neurobiol Dis 2003, 14:81-88.
• [18]Ohno M, Cole SL, Yasvoina M, Zhao J, Citron M, Berry R, Disterhoft JF, Vassar R: BACE1 gene deletion prevents neuron loss and memory deficits in 5XFAD APP/PS1 transgenic mice. Neurobiol Dis 2007, 26:134-145.
• [19]Laird FM, Cai H, Savonenko AV, Farah MH, He K, Melnikova T, Wen H, Chiang HC, Xu G, Koliatsos VE, et al.: BACE1, a major determinant of selective vulnerability of the brain to amyloid-beta amyloidogenesis, is essential for cognitive, emotional, and synaptic functions. J Neurosci 2005, 25:11693-11709.
• [20]McConlogue L, Buttini M, Anderson JP, Brigham EF, Chen KS, Freedman SB, Games D, Johnson-Wood K, Lee M, Zeller M, et al.: Partial reduction of BACE1 has dramatic effects on Alzheimer plaque and synaptic pathology in APP Transgenic Mice. J Biol Chem 2007, 282:26326-26334.
• [21]Gouras GK, Almeida CG, Takahashi RH: Intraneuronal Abeta accumulation and origin of plaques in Alzheimer’s disease. Neurobiol Aging 2005, 26:1235-1244.
• [22]LaFerla FM, Green KN, Oddo S: Intracellular amyloid-beta in Alzheimer’s disease. Nat Rev Neurosci 2007, 8:499-509.
• [23]Wirths O, Erck C, Martens H, Harmeier A, Geumann C, Jawhar S, Kumar S, Multhaup G, Walter J, Ingelsson M, et al.: Identification of low molecular weight pyroglutamate A{beta} oligomers in Alzheimer disease: a novel tool for therapy and diagnosis. J Biol Chem 2010, 285:41517-41524.
• [24]Youmans KL, Tai LM, Nwabuisi-Heath E, Jungbauer L, Kanekiyo T, Gan M, Kim J, Eimer WA, Estus S, Rebeck GW, et al.: APOE4-specific changes in Abeta accumulation in a new transgenic model of Alzheimer’s disease. J Biol Chem 2012, 287:41774-41786.
• [25]Youmans KL, Tai LM, Kanekiyo T, Stine WB Jr, Michon SC, Nwabuisi-Heath E, Manelli AM, Fu Y, Riordan S, Eimer WA, et al.: Intraneuronal Abeta detection in 5xFAD mice by a new Abeta-specific antibody. Mol Neurodegener 2012, 7:8. BioMed Central Full Text
• [26]Moon M, Hong HS, Nam DW, Baik SH, Song H, Kook SY, Kim YS, Lee J, Mook-Jung I: Intracellular amyloid-beta accumulation in calcium-binding protein-deficient neurons leads to amyloid-beta plaque formation in animal model of Alzheimer’s disease. J Alzheimers Dis 2012, 29:615-628.
• [27]Oddo S, Caccamo A, Smith IF, Green KN, LaFerla FM: A dynamic relationship between intracellular and extracellular pools of Abeta. Am J Pathol 2006, 168:184-194.
• [28]Winton MJ, Lee EB, Sun E, Wong MM, Leight S, Zhang B, Trojanowski JQ, Lee VM: Intraneuronal APP, not free Abeta peptides in 3xTg-AD mice: implications for tau versus Abeta-mediated Alzheimer neurodegeneration. J Neurosci 2011, 31:7691-7699.
• [29]Turner RS, Suzuki N, Chyung AS, Younkin SG, Lee VM: Amyloids beta40 and beta42 are generated intracellularly in cultured human neurons and their secretion increases with maturation. J Biol Chem 1996, 271:8966-8970.
• [30]Friedrich RP, Tepper K, Ronicke R, Soom M, Westermann M, Reymann K, Kaether C, Fandrich M: Mechanism of amyloid plaque formation suggests an intracellular basis of Abeta pathogenicity. Proc Natl Acad Sci USA 2010, 107:1942-1947.
• [31]Langui D, Girardot N, El Hachimi KH, Allinquant B, Blanchard V, Pradier L, Duyckaerts C: Subcellular topography of neuronal Abeta peptide in APPxPS1 transgenic mice. Am J Pathol 2004, 165:1465-1477.
• [32]Takahashi RH, Milner TA, Li F, Nam EE, Edgar MA, Yamaguchi H, Beal MF, Xu H, Greengard P, Gouras GK: Intraneuronal Alzheimer abeta42 accumulates in multivesicular bodies and is associated with synaptic pathology. Am J Pathol 2002, 161:1869-1879.
• [33]Zhang Y, McLaughlin R, Goodyer C, LeBlanc A: Selective cytotoxicity of intracellular amyloid beta peptide1-42 through p53 and Bax in cultured primary human neurons. J Cell Biol 2002, 156:519-529.
• [34]Rohn TT, Head E: Caspase activation in Alzheimer’s disease: early to rise and late to bed. Rev Neurosci 2008, 19:383-393.
• [35]LeBlanc AC: The role of apoptotic pathways in Alzheimer’s disease neurodegeneration and cell death. Curr Alzheimer Res 2005, 2:389-402.
• [36]Behl C: Apoptosis and Alzheimer’s disease. J Neural Transm 2000, 107:1325-1344.
• [37]Christensen DZ, Kraus SL, Flohr A, Cotel MC, Wirths O, Bayer TA: Transient intraneuronal A beta rather than extracellular plaque pathology correlates with neuron loss in the frontal cortex of APP/PS1KI mice. Acta Neuropathol 2008, 116:647-655.
• [38]Schmitz C, Rutten BP, Pielen A, Schafer S, Wirths O, Tremp G, Czech C, Blanchard V, Multhaup G, Rezaie P, et al.: Hippocampal neuron loss exceeds amyloid plaque load in a transgenic mouse model of Alzheimer’s disease. Am J Pathol 2004, 164:1495-1502.
• [39]Almeida CG, Takahashi RH, Gouras GK: Beta-amyloid accumulation impairs multivesicular body sorting by inhibiting the ubiquitin-proteasome system. J Neurosci 2006, 26:4277-4288.
• [40]Tseng BP, Green KN, Chan JL, Blurton-Jones M, LaFerla FM: Abeta inhibits the proteasome and enhances amyloid and tau accumulation. Neurobiol Aging 2008, 29:1607-1618.
• [41]Kasa P Sr, Papp H, Kasa P Jr, Pakaski M, Balaspiri L: Effects of amyloid-beta on cholinergic and acetylcholinesterase-positive cells in cultured basal forebrain neurons of embryonic rat brain. Brain Res 2004, 998:73-82.
• [42]Gastard MC, Troncoso JC, Koliatsos VE: Caspase activation in the limbic cortex of subjects with early Alzheimer’s disease. Ann Neurol 2003, 54:393-398.
• [43]Hwang DY, Chae KR, Kang TS, Hwang JH, Lim CH, Kang HK, Goo JS, Lee MR, Lim HJ, Min SH, et al.: Alterations in behavior, amyloid beta-42, caspase-3, and Cox-2 in mutant PS2 transgenic mouse model of Alzheimer’s disease. FASEB J 2002, 16:805-813.
• [44]Uetsuki T, Takemoto K, Nishimura I, Okamoto M, Niinobe M, Momoi T, Miura M, Yoshikawa K: Activation of neuronal caspase-3 by intracellular accumulation of wild-type Alzheimer amyloid precursor protein. J Neurosci 1999, 19:6955-6964.
• [45]Kim HS, Lee JH, Lee JP, Kim EM, Chang KA, Park CH, Jeong SJ, Wittendorp MC, Seo JH, Choi SH, Suh YH: Amyloid beta peptide induces cytochrome C release from isolated mitochondria. Neuroreport 2002, 13:1989-1993.
• [46]Stadelmann C, Deckwerth TL, Srinivasan A, Bancher C, Bruck W, Jellinger K, Lassmann H: Activation of caspase-3 in single neurons and autophagic granules of granulovacuolar degeneration in Alzheimer’s disease. Evidence for apoptotic cell death. Am J Pathol 1999, 155:1459-1466.
• [47]Su JH, Zhao M, Anderson AJ, Srinivasan A, Cotman CW: Activated caspase-3 expression in Alzheimer’s and aged control brain: correlation with Alzheimer pathology. Brain Res 2001, 898:350-357.
• [48]Yang DS, Kumar A, Stavrides P, Peterson J, Peterhoff CM, Pawlik M, Levy E, Cataldo AM, Nixon RA: Neuronal apoptosis and autophagy cross talk in aging PS/APP mice, a model of Alzheimer’s disease. Am J Pathol 2008, 173:665-681.
• [49]D’Amelio M, Cavallucci V, Middei S, Marchetti C, Pacioni S, Ferri A, Diamantini A, De Zio D, Carrara P, Battistini L, et al.: Caspase-3 triggers early synaptic dysfunction in a mouse model of Alzheimer’s disease. Nat Neurosci 2011, 14:69-76.