, Volume 29, Issue 2–3, pp 87–96

Neuropathological quantification of dtg APP/PS1: neuroimaging, stereology, and biochemistry

  • Kebreten F. Manaye
  • Paul C. Wang
  • Jahn N. O’Neil
  • Sophia Y. Huang
  • Tao Xu
  • De-Liang Lei
  • Yousef Tizabi
  • Mary Ann Ottinger
  • Donald K. Ingram
  • Peter R. Mouton


Murine models that mimic the neuropathology of Alzheimer’s disease (AD) have the potential to provide insight into the pathogenesis of the disease and lead to new strategies for the therapeutic management of afflicted patients. We used magnetic resonance imaging (MRI), design-based stereology, and high performance liquid chromatography (HPLC) to assess the age-related neuropathology in double transgenic mice that overexpress two AD-related proteins—amyloid precursor protein (APP) and presenilin 1 (PS1)—and age- and gender-matched wild-type (WT) controls. In mice ranging in age from 4–28 months, total volumes of the hippocampal formation (VHF) and whole brain (Vbrain) were quantified by the Cavalieri-point counting method on a systematic-random sample of coronal T2-weighted MRI images; the same stereological methods were used to quantify VHF and Vbrain after perfusion and histological processing. To assess changes in AD-type beta-amyloid (Aβ) plaques, sections from the hippocampal formation and amylgdaloid complex of mice aged 5, 12, and 15 months were stained by Congo Red histochemistry. In aged mice with large numbers of amyloid plaques, systematic-random samples of sections were stained by GFAP immunocytochemistry to assess gender and genotype effects on total numbers of astrocytes. In addition, levels of norepinephrine (NE), dopamine (DA), serotonin (5-HT) and 5-HT metabolites were assayed by HPLC in fresh-frozen samples from neocortex, striatum, hippocampus, and brainstem. We confirmed age-related increases in amyloid plaques, beginning with a few plaques at 5 months of age and increasing densities by 12 and 15 months. At 15 months of age, there were robust genotype effects, but no gender effects, on GFAP-immunopositive astrocytes in the amygdaloid complex and hippocampus. There were no effects on monoamine levels in all brain regions examined, and no volume changes in hippocampal formation or whole brain as quantified on either neuroimages or tissue sections. Strong correlations were present between volume estimates from MRI images and histological sections, with about 85% reduction in mean VHF or mean Vbrain between MRI and processed histological sections. In summary, these findings show that the double transgenic expression of AD-type mutations is associated with age-related increases in amyloid plaques and astrocytosis; however, this model does not recapitulate the cortical atrophy or neurochemical changes that are characteristic of AD.


MRI Alzheimer’s disease Hippocampal formation Amygdala Unbiased stereology 


  1. Aletrino MA, Vogels OJ, Van Domburg PH, Ten Donkelaar HJ (1992) Cell loss in the nucleus raphe dorsalis in Alzheimer’s disease. Neurobiol Aging 13(4):461–468PubMedCrossRefGoogle Scholar
  2. Alzheimer A (1907) Ueber eine eigenartige Erkrankung der Hirnrinde. Allgemeine Zeitschrift fur Psychiatrie 64:146–148Google Scholar
  3. Benveniste H, Einstein G, Kim KR, Hulette C, Johnson GA (1999) Detection of neuritic plaques in Alzheimer’s disease by magnetic resonance microscopy. Proc Natl Acad Sci USA 96(24):14079–14084PubMedCrossRefGoogle Scholar
  4. Borthakur A, Urya K, Chively SB, Poptani H, Corbo M, Charagundla SR, Trojanoqski JQ, Lee VM, Reddy R (2003) In Vivo T1 weighted MRI of amyloid transgenic mouse model of Alzheimer’s disease. Proc Intl Soc Mag Reson Med 11:2039Google Scholar
  5. Cavalieri B (1635) Geometria indivibilibus continuorum. Bononi: Typis Clementis Ferronij. Reprint from (1966) Geometria degli indivisibili. Torino: Unione Tipografico-Editrice TorineseGoogle Scholar
  6. Convit A, de Leon MJ, Golomb J, George AE, Tarshish CY, Bobinski M, Tsui W, De Santi S, Wegiel J, Wisniewski H (1993) Hippocampal atrophy in early Alzheimer’s disease: anatomic specificity and validation. Psychiatr Q 64:371–387PubMedCrossRefGoogle Scholar
  7. Dedeoglu A, Choi JK, Cormier K, Kowall NW, Jenkins BG (2004) Magnetic resonance spectroscopic analysis of Alzheimer’s disease mouse brain that express mutant APP shows altered neurochemical profile. Brain Res 1012(1–2):60–65PubMedCrossRefGoogle Scholar
  8. DeKosky ST, Scheff SW (1990) Synapse loss in frontal cortex biopsies in Alzheimer’s disease: correlation with cognitive severity. Ann Neurol 27:457–464PubMedCrossRefGoogle Scholar
  9. de la Monte SM (1989) Quantitation of cerebral atrophy in preclinical and end-stage Alzheimer’s disease. Ann Neurol 25:450–459PubMedCrossRefGoogle Scholar
  10. de Leon MJ, DeSanti S, Zinkowski R, Mehta PD, Pratico D, Segal S, Clark C, Kerkman D, DeBernardis J, Li J, Lair L, Reisberg B, Tsui W, Rusinek H (2004) MRI and CSF studies in the early diagnosis of Alzheimer’s disease. J Intern Med 256:205–223PubMedCrossRefGoogle Scholar
  11. Gunderson HJ, Jensen EB (1987) The efficiency of systematic sampling in stereology and its prediction. J Microsc 147(Pt 3):229–263Google Scholar
  12. Gundersen HJG, Jensen EV, Kieu K, Nielsen J (1999) The efficiency of systematic sampling in stereology - revisited. J Microsc 193:199–211PubMedCrossRefGoogle Scholar
  13. Gundersen HJG, Østerby R (1981) Optimizing sampling efficiency of stereological studies in biology: or “Do more less well.” J Microsc 121:65–73PubMedGoogle Scholar
  14. Helpern JA, Lee S-P, Falangol MF, Dyakin VV, Bogart A, Ardekani B, Duff K, Branc C, Wisniewski T, de Leon MJ, Wolf O, O’Shea J, Nixon RA (2004) MRI assessment of neuropathology in a transgenic mouse model of Alzheimer’s disease. Magn Reson Med 51:794–798PubMedCrossRefGoogle Scholar
  15. Jankowsky J, Fadale DJ, Andersen JK, Xu G, Gonzales V, Jenkins NA, Copeland NG, Lee MK, Younkin LH, Wagner SL, Younkin SG, Borchelt DR (2003) 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):159–170PubMedCrossRefGoogle Scholar
  16. Jobst KA, Smith AD, Szatmari M, Esiri MM, Jaskowski A, Hindley N, McDonald B, Molyneux AJ (1994) Rapidly progressing atrophy of medial temporal lobe in Alzheimer’s disease. Lancet 343:829–830PubMedCrossRefGoogle Scholar
  17. Lee GD, Aruna JH, Barrett JM, Lei D-L, Ingram DK, Mouton PR (2005) Stereological analysis of microvascular parameters in a double transgenic model of Alzheimer’s disease. Brain Res Bull 65:317–322PubMedCrossRefGoogle Scholar
  18. Long JM, Kalehua AN, Muth NJ, Calhoun ME, Jucker M, Hengemihle JM, Ingram DK, Mouton PR (1998) Stereological analysis of astrocyte and microglia in aging mouse hippocampus. Neurobiol Aging 19:497–503PubMedCrossRefGoogle Scholar
  19. McGowan E, Pickford F, Dickson DW (2003) Alzheimer animal models: models of Aβ deposition in transgenic mice. In: Dickson DW (ed) Neurodegeneration: the molecular pathology of dementia and movement disorders. ISN Neuropath Press, Basel, pp 74–79Google Scholar
  20. McKeel DW, Price JL, Miller JP, Grant EA, Xiong C, Berg L, Morris JC (2004) Neuropathologic criteria for diagnosing Alzheimer disease in persons with pure dementia of Alzheimer type. J Neuropathol Exp Neurol 63(10):1028–1037PubMedGoogle Scholar
  21. Mirra S, Hart MH, Terry RD (1993) Making the diagnosis of Alzheimer’s disease. A primer for practicing pathologists. Arch Pathol Lab Med 117:132–144PubMedGoogle Scholar
  22. Mouton PR (2002) Principles and practices of unbiased stereology: an introduction for bioscientists. Johns Hopkins University Press, BaltimoreGoogle Scholar
  23. Mouton PR, Long JM, Lei DL, Howard V, Jucker M, Calhoun ME, Ingram DK (2002) Age and gender effects on microglia and astrocyte numbers in brains of mice. Brain Res 956(1):30–35PubMedCrossRefGoogle Scholar
  24. Mouton PR, Martin LJ, Calhoun ME, Dal Forno G, Price DL (1998) Cognitive decline strongly correlates with cortical atrophy in Alzheimer’s dementia. Neurobiol Aging 19:371–377PubMedCrossRefGoogle Scholar
  25. Mouton PR, Pakkenberg B, Gundersen HJG, Price DL (1994) Absolute number and size of pigmented locus coeruleus neurons in the brains of young and aged individuals. J Chem Neuroanat 7:185–190PubMedCrossRefGoogle Scholar
  26. Mouton PR, Price DL, Walker LC (1997) Empirical assessment of synapse numbers in primate neocortex. J Neurosci Methods 75:119–126PubMedCrossRefGoogle Scholar
  27. Ohno M, Chang L, Tseng W, Oakley H, Citron M, Klein WL, Vassar R, Disterhoft JF (2006) Temporal memory deficits in Alzheimer’s mouse models: rescue by genetic deletion of BACE1. Eur J Neurosci 23(1):251–260PubMedCrossRefGoogle Scholar
  28. O’Neil JN, et al. Catecholaminergic neuronal loss in locus coeruleus of aged female dtg APP/PS1 mice. J Chem Neuroanat (in press)Google Scholar
  29. Poduslo JF, Wengenack TM, Curran GL, Wisniewski T, Sigurdsson EM, Macura SI, Borowski BJ, Jack CR Jr (2002) Molecular targeting of Alzheimer’s amyloid plaques for contrast-enhanced magnetic resonance imaging. Neurobiol Dis 11(2):315–329PubMedCrossRefGoogle Scholar
  30. Roberts N, Puddephat MJ, McNulty V (2000) The benefit of stereology for quantitative radiology. Br J Radiol 73(871):679–697PubMedGoogle Scholar
  31. Savenenko A, Xu GM, Melnikova T, Morton J, Gonzales V, Wong M, Price DL, Tang F, Markowska AL, Borchelt DR (2005) Episodic-like memory deficits in the APPswe/PS1dE9 mouse model of Alzheimer’s disease: relationship to β-amyloid deposition and neurotransmitter abnormalities. Neurobiol Dis 18(3):602–617CrossRefGoogle Scholar
  32. Storga D, Vrecko K, Birkmayer JG, Reibneggar G, Cavalieri B (1996) Monoaminergic neurotransmitters, their precursors and metabolites in brains of Alzheimer patients. Neurosci Lett 203(1):29–32PubMedCrossRefGoogle Scholar
  33. Stout JC, Jernigan TL, Archibald SL, Salmon DP (1996) Association of dementia severity with cortical gray matter and abnormal white matter volumes in dementia of the Alzheimer type. Arch Neurol 53:742–749PubMedGoogle Scholar
  34. Subbiah P, Mouton PR, Fedor H, McArthur JC, Glass JD (1996) Stereological analysis of cerebral atrophy in human immunodeficiency virus-associated dementia. Exp Neurol 55(10):1032–1037Google Scholar
  35. Szapacs ME, Numis AL, Andrews AM (2004) Late onset loss of hippocampal 5-HT and NE is accompanied by increases in BDNF protein expression in mice co-expressing mutant APP and PS1. Neurobiol Dis 16(3):572–580PubMedCrossRefGoogle Scholar
  36. Sze CI, Troncoso JC, Kawas C, Mouton PR, Price DL, Martin LJ (1997) Loss of the presynaptic vesicle protein synaptophysin in hippocampus correlates with cognitive decline in Alzheimer disease. J Neuropathol Exp Neurol 56(8):933–944PubMedCrossRefGoogle Scholar
  37. Terry RD, Masliah E, Salmon DP, Butters N, DeTeresa R, Hill R, Hansen LA, Katzman R (1991) Physical basis of cognitive alterations in Alzheimer’s disease: synapse loss is the major correlate of cognitive impairment. Ann Neurol 30:572–580PubMedCrossRefGoogle Scholar
  38. Tiraboschi P, Hansen LA, Thal LJ, Corey-Bloom J (2004) The importance of neuritic plaques and tangles to the development and evolution of AD. Neurology 64:1984–1989Google Scholar
  39. Tuppo EE, Arias HR (2005) The role of inflammation in Alzheimer’s disease. Int J Biochem Cell Biol 37(2):289–305PubMedCrossRefGoogle Scholar
  40. Wadghirii YZ, Sigurdsson EM, Sadowski M, Elliott JI, Li Y, Scholtzova H, Tang Cy, Aguinaldo G, Pappolla M, Duff K, Wisniewski T, Turnbull DH (2003) Detection of Alzheimer’s amyloid in transgenic mice using magnetic resonance microimaging. Magn Reson Med 50(2):293–302CrossRefGoogle Scholar
  41. West MJ (1993) Regionally specific loss of neurons in the aging human hippocampus. Neurobiol Aging 14(4):287–293PubMedCrossRefGoogle Scholar
  42. Zarow C, Lynness SA, Mortimer JA, Chui HC (2003) Neuronal loss is greater in the locus coerulus than nucleus basalis and substantia nigra in Alzheimer and Parkinson diseases. Arch Neurol 60(3):337–341PubMedCrossRefGoogle Scholar

Copyright information

© American Aging Association, Media, PA, USA 2007

Authors and Affiliations

  • Kebreten F. Manaye
    • 1
  • Paul C. Wang
    • 2
  • Jahn N. O’Neil
    • 1
    • 8
  • Sophia Y. Huang
    • 2
  • Tao Xu
    • 1
  • De-Liang Lei
    • 1
  • Yousef Tizabi
    • 3
  • Mary Ann Ottinger
    • 4
    • 5
  • Donald K. Ingram
    • 5
    • 6
  • Peter R. Mouton
    • 7
  1. 1.Department of Physiology and BiophysicsHoward University College of MedicineWashingtonUSA
  2. 2.Department of RadiologyHoward University College of MedicineWashingtonUSA
  3. 3.Department of PharmacologyHoward University College of MedicineWashingtonUSA
  4. 4.University of MarylandCollege ParkUSA
  5. 5.Laboratory of Experimental GerontologyNational Institute on Aging, NIHBaltimoreUSA
  6. 6.Nutritional Neuroscience and Aging LaboratoryLouisiana State UniversityBaton RougeUSA
  7. 7.Stereology Resource CenterChesterUSA
  8. 8.School of MedicineJohns Hopkins UniversityBaltimoreUSA

Personalised recommendations