Pyroglutamate-3 Amyloid-β Deposition in the Brains of Humans, Non-Human Primates, Canines, and Alzheimer Disease–Like Transgenic Mouse Models

• DOI: 10.1016/j.ajpath.2013.05.005
• Publication Date: 2013-06-07T06:48:00Z
• Authors: Jeffrey L. Frost; Kevin X. Le; Holger Cynis; Elizabeth Ekpo; Martin Kleinschmidt; Roberta M. Palmour; Frank R. Ervin; Shikha Snigdha; Carl W. Cotman; Takaomi C. Saido; Robert J. Vassar; Peter St. George-...; Tsuneya Ikezu; Stephan Schilling; Hans-Ulrich Demuth; Cynthia A. Lemere
• Keywords: Amyloid (mycology); Human disease

Amyloid-β (Aβ) peptides, starting with pyroglutamate at the third residue (pyroGlu-3 Aβ), are a major species deposited in brain of Alzheimer disease (AD) patients. Recent studies suggest that this isoform shows higher toxicity and amyloidogenecity when compared to full-length Aβ peptides. Here, we report first comprehensive comparative IHC evaluation pyroGlu-3 deposition humans animal models. PyroGlu-3 immunoreactivity (IR) is abundant plaques cerebral amyloid angiopathy AD Down syndrome patients, colocalizing general IR. further present two nontransgenic mammalian models amyloidosis, Caribbean vervets, beagle canines. In addition, was analyzed 12 different AD-like transgenic mouse contrast humans, all showed preceding deposition. 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Raju Greenberg Bacskai Frosch Characterization APPswe/PS1dE9 24: 516-524Crossref (559) Specific mutations used generate illustrated 2.Table 2Semiquantitative (0 4+) Scoring General Immunoreactive Plaques Aβ–Positive AD-Like Mouse Models Examined modelReferencePromoterAge (months)R1282 IR6E10 IRJ20 C57BL/6 APPK670N/M671L,V717F29Mucke ScholarPDGF8 (n = 2)2+1+2+12 6)2.5+2.67+3+17 4)2.75+2.25+2.75+30 2)4+3+3+J20 B6D2F1 APPK670N/M671L,V717F30Shankar ScholarPDGF6 4)2.5+1.75+1.75+9 4)2+0.75+1.5+16 5)4+3+3+24 2)4+3+3+J20;C3−/− APPK670N/M671L,V717F factor deficient31Maier 6)2.17+1.5+2.17+12 10)2.8+1.9+3+17 5)4+3+3+hAPP751 APPK670N/M671L,V717I32Rockenstein ScholarmThy-114 2)4+4+3+Tg2576 Tg2576/CCL2 APPK670N/M671L,V717I33Hsiao 34Yamamoto ScholarHamster PrP hGFAP (CCL2)14 4)2+1.25+2+14 4)2.75+1.75+2.5+TgSwDI APPK670N/M671L E693Q, D694N35Davis ScholarmThy-13 4)1.5+0.75+2.25+6 2)3+2+1.5+12 4)3+1.5+3+24 4)4+3+3+TgCRND8 APPK670N/M671L/V717F36Chishti PrP83 days 3)2+0.5+2+<300 2)4+3+3+3xTg-AD APPK670N/M671L, PS1M146V ,TauP301L37Oddo ScholarmThy-1 (APP, Tau) PS1 KI≥14 8)1+0.5+1+22 4)2+1+2+≤25 3)3+1.5+3+5xFAD I716V, V717I PS1M146L, L286V38Oakley ScholarmThy-11.5 1)1+0NE2 2)2.5+0NE4 1)4+2+NE6 2)4+3+NE9 1)4+3+NE12 12)4+4+NEPSAPP APPK670N/M671Lx PS1M146L39Holcomb PDGF (PS1)4 6)2+0NE18 4)4+4+3+APP/PS1ΔE9 APPK595N/M596L deletion exon 940Garcia-Alloza ScholarmPrP PS1)6 2)2+1+2+14 4)4+4+3+General confirmed anti-Aβ mAb 6E10. Plaque scoring each group given mean value calculated sum individual scores divided number animals per group. key: 1+ 5 plaques; 2+ 6 plaques, 3+ 100 4+ 101 more plaques.NE, not examined. plaques. NE, blocks 10% neutral buffered hours (brief fixation) exception one years (summarized 1). Archived, formalin-fixed primate brains blocked nine regions (rostrocaudal). Tissue processed paraffin embedding then sectioned μm analysis. Canines euthanized Irvine, sodium pentobarbital overdose, removed, left hemisphere 4% paraformaldehyde months, blocked, embedding, μm. Mice anesthetized CO2 inhalation, perfused 20 mL ice-cold saline followed removal. Hemibrains 24 before being All use Harvard Medical Area Standing Animals compliance state federal regulations. Aβ–specific clone characterized surface plasmon resonance. Briefly, performed Biacore 3000 (GE Healthcare, Chalfont St Giles, UK). CM5 chip covalently conjugated response units (RU) test flow cells (Fc) 2, 3, 4 (Supplemental S1). Fc prepared blank subtraction. Clone diluted running buffer HEPES-buffered saline, mmol/L EDTA, 0.005% v/v surfactant P20 (HBS-EP) concentration 25 μg/mL. First, basal