Building a More Perfect Beast: APP Transgenic Mice with Neuronal Loss

Abstract

While there have been a number of successful attempts to develop transgenic mice that express mutant amyloid precursor protein (APP) and develop amyloid deposits that have features similar to those in aging and Alzheimer's disease (AD),1McGowan E Pickford F Dickson DW Alzheimer animal models: models of Aβ deposition in transgenic mice.in: Dickson DW Neurodegeneration: The Molecular Pathology of Dementia and Movement Disorders. ISN Neuropath Press, Basel2003: 74-79Google Scholar until now there has been either no neuronal loss or less than convincing evidence in favor of neuronal loss. In this issue of The American Journal of Pathology, Schmitz and colleagues2Schmitz C Rutten BPF Pielen A Schäfer S Wirths O Tremp G Czech C Blanchard V Multhaup G Rezaie P Korr H Steinbusch HWM Pradier L Bayer TA Hippocampal neuronal loss exceeds amyloid plaque load in a transgenic mouse model of Alzheimer's disease.Am J Pathol. 2004; 164: 1495-1502Abstract Full Text Full Text PDF PubMed Scopus (221) Google Scholar provide compelling evidence for age-dependent neuronal loss in the hippocampus of bigenic transgenic mice expressing a double mutation in APP (Swedish APP mutation APPK670N, M671L (APPsw) and APPV717I, under the mouse Thy1 promoter) and mutant presenilin-1 (PS-1 M146L under the pHMG promoter). The neuronal loss was ascertained by unbiased stereologic methods on Nissl-stained sections. The animals showed significant (30 to 35%) neuronal loss in the hippocampal pyramidal layer compared to controls, which was disproportionate to the degree of amyloid deposition. No neuronal loss was detected in the dentate fascia of the hippocampus, the only other region assessed for neuronal loss. While the regions analyzed are of interest, if the model is to have relevance to AD, it will be important in future studies to extend neuronal counts to limbic and association cortical regions, which are especially vulnerable to amyloid deposition in humans. The behavioral consequences, if any, of neuronal loss in this model are not described in this report or other descriptions of this model3Blanchard V Moussaoui S Czech C Touchet N Bonici B Planche M Canton T Jedidi I Gohin M Wirths O Bayer TA Langui D Duyckaerts C Tremp G Pradier L Time sequence of maturation of dystrophic neurites associated with Aβ deposits in APP/PS1 transgenic mice.Exp Neurol. 2003; 184: 247-263Crossref PubMed Scopus (248) Google Scholar and are left to future work. The mechanism of neuronal loss remains to be determined, but previous studies of this model demonstrated intra-neuronal Aβ as well as neuronal expression of oxidative stress and pro-apoptotic markers.3Blanchard V Moussaoui S Czech C Touchet N Bonici B Planche M Canton T Jedidi I Gohin M Wirths O Bayer TA Langui D Duyckaerts C Tremp G Pradier L Time sequence of maturation of dystrophic neurites associated with Aβ deposits in APP/PS1 transgenic mice.Exp Neurol. 2003; 184: 247-263Crossref PubMed Scopus (248) Google Scholar This would suggest that mechanisms other than those associated with extracellular amyloid deposits are involved. The poor correlation of neuronal loss with extracellular amyloid deposits fits with previous studies of humans and other animal models of AD. Amyloid burden, as measured by image analysis or point counting methods as in this study, has not proven to be the most robust correlate with cognitive impairment and neuronal loss in humans4Gomez-Isla T Hollister R West H Mui S Growdon JH Petersen RC Parisi JE Hyman BT Neuronal loss correlates with but exceeds neurofibrillary tangles in Alzheimer's disease.Ann Neurol. 1997; 41: 17-24Crossref PubMed Scopus (1093) Google Scholar or with behavioral abnormalities in transgenic models of AD.5Holcomb LA Gordon MN Jantzen P Hsiao K Duff K Morgan D Behavioral changes in transgenic mice expressing both amyloid precursor protein and presenilin-1 mutations: lack of association with amyloid deposits.Behav Genet. 1999; 29: 177-185Crossref PubMed Scopus (306) Google Scholar In contrast, cognitive dysfunction correlates better with markers of neuronal degeneration, such a synaptic loss6Terry RD Masliah E Salmon DP Butters N DeTeresa R Hill R Hansen LA Katzman R Physical basis of cognitive alterations in Alzheimer's disease: synapse loss is the major correlate of cognitive impairment.Ann Neurol. 1991; 30: 572-580Crossref PubMed Scopus (3287) Google Scholar or neurofibrillary pathology.7Bierer LM Hof PR Purohit DP Carlin L Schmeidler J Davis KL Perl DP Neocortical neurofibrillary tangles correlate with dementia severity in Alzheimer's disease.Arch Neurol. 1995; 52: 81-88Crossref PubMed Scopus (396) Google Scholar Given the popular belief that amyloid is the cause of neuronal degeneration in AD, these findings have prompted modification of the original “amyloid cascade” hypothesis for AD.8Hardy J Selkoe DJ The amyloid hypothesis of Alzheimer's disease: progress and problems on the road to therapeutics.Science. 2002; 297: 353-356Crossref PubMed Scopus (10646) Google Scholar Recent research has changed its focus to possible toxic effects of protofibrillar forms of Aβ or intra-neuronal Aβ. The latter is a feature described in this model as well as other AD models that have shown cognitive and synaptic dysfunction.9Oddo S Caccamo A Shepherd JD Murphy MP Golde TE Kayed R Metherate R Mattson MP Akbari Y LaFerla FM Triple-transgenic model of Alzheimer's disease with plaques and tangles: intracellular Aβ and synaptic dysfunction.Neuron. 2003; 39: 409-421Abstract Full Text Full Text PDF PubMed Scopus (3059) Google Scholar Unfortunately, the anatomical relationship between neurons with cytoplasmic Aβ and neuronal loss is not described, albeit the neurons with cytoplasmic Aβ are described as being restricted in number and distribution.3Blanchard V Moussaoui S Czech C Touchet N Bonici B Planche M Canton T Jedidi I Gohin M Wirths O Bayer TA Langui D Duyckaerts C Tremp G Pradier L Time sequence of maturation of dystrophic neurites associated with Aβ deposits in APP/PS1 transgenic mice.Exp Neurol. 2003; 184: 247-263Crossref PubMed Scopus (248) Google Scholar One would hope that if intra-neuronal Aβ is detrimental to long-term neuronal survival, it should antedate and parallel the distribution of neuron loss. In addition to those mentioned, a number of other areas of investigation remain to establish this bigenic mouse as a valuable model for AD. Given the evidence that synaptic loss is an important structural correlate of cognitive impairment in AD, it is important that synaptic integrity be assessed in future studies. The time course of neuronal and potential synaptic loss may shed light on the mechanism of synaptic degeneration in AD. The role of neuro-inflammation in neuronal loss also is an active area of investigation, and the anatomical and temporal relationships of inflammatory reaction associated with amyloid deposition and neuronal loss will be important to determine. Finally, a detailed time course of changes in the levels of various Aβ species may also shed light on the mechanism of neuronal loss, as previous studies have shown a predictable sequence of changes in Aβ solubility that antedate visible extracellular deposits.10Kawarabayashi T Younkin LH Saido TC Shoji M Ashe KH Younkin SG Age-dependent changes in brain CSF, and plasma amyloid (β) protein in the Tg2576 transgenic mouse model of Alzheimer's disease.J Neurosci. 2001; 21: 372-381Crossref PubMed Google Scholar The question that arises from this report is why this model is associated with neuronal loss, while previous APP models are not. There are a number of factors to consider. The genetic constructs used to produce the mice are the most obvious factor. The precise combination of transgenes and promoters in this animal has not been used in any of the other reported APP models of AD. The APP mutation is a double mutation (APPsw and APPV717I) on APP751. APP751 is a form of APP that is found in neurons, but also in glial cells.11de Silva RHA Jen A Wickenden C Jen LS Wilkinson SL Patel AJ Cell-specific expression of β-amyloid precursor protein isoform mRNAs and proteins in neurons and astrocytes.Brain Res Mol Brain Res. 1997; 47: 147-156Crossref PubMed Scopus (93) Google Scholar Most APP models have used APP695.1McGowan E Pickford F Dickson DW Alzheimer animal models: models of Aβ deposition in transgenic mice.in: Dickson DW Neurodegeneration: The Molecular Pathology of Dementia and Movement Disorders. ISN Neuropath Press, Basel2003: 74-79Google Scholar The first APP model with convincing amyloid pathology was PDAPP mice,12Games D Adams D Alessandrini R Barbour R Berthelette P Blackwell C Carr T Clemens J Donaldson T Gillespie F Guido T Hagopian S Johnson-Wood K Khan K Lee M Leibowitz P Lieberburg I Little S Masliah E McConlogue L Montoya-Zavala M Mucke L Paganini L Penniman E Power M Schenk D Seubert P Snyder B Soriano F Tan H Vitale J Wadsworth S Wolozin B Zhao J Alzheimer-type neuropathology in transgenic mice overexpressing V717F β-amyloid precursor protein.Nature. 1995; 373: 523-527Crossref PubMed Scopus (2217) Google Scholar which has a different APP mutation, APPV717F; a different promoter, PDGF; and a construct that leads to expression of APP751, APP770, and APP695 rather than just APP751. The PDAPP mice develop amyloid deposits as early as 6 months of age, and the plaques have many features in common with those seen in aging and Alzheimer's disease, including dystrophic neurites and reactive glia. Initial studies failed to find significant neuronal loss.13Irizarry MC Soriano F McNamara M Page KJ Schenk D Games D Hyman BT Aβ deposition is associated with neuropil changes, but not with overt neuronal loss in the human amyloid precursor protein V717F (PDAPP) transgenic mouse.J Neurosci. 1997; 17: 7053-7059Crossref PubMed Google Scholar In contrast, the model reported by Schmitz and co-workers2Schmitz C Rutten BPF Pielen A Schäfer S Wirths O Tremp G Czech C Blanchard V Multhaup G Rezaie P Korr H Steinbusch HWM Pradier L Bayer TA Hippocampal neuronal loss exceeds amyloid plaque load in a transgenic mouse model of Alzheimer's disease.Am J Pathol. 2004; 164: 1495-1502Abstract Full Text Full Text PDF PubMed Scopus (221) Google Scholar, 3Blanchard V Moussaoui S Czech C Touchet N Bonici B Planche M Canton T Jedidi I Gohin M Wirths O Bayer TA Langui D Duyckaerts C Tremp G Pradier L Time sequence of maturation of dystrophic neurites associated with Aβ deposits in APP/PS1 transgenic mice.Exp Neurol. 2003; 184: 247-263Crossref PubMed Scopus (248) Google Scholar has amyloid deposits as early as 2.5 months of age. The PDAPP mice have evidence of perturbation of neuronal populations in the vicinity of amyloid deposits,14Le R Cruz L Urbanc B Knowles RB Hsiao-Ashe K Duff K Irizarry MC Stanley HE Hyman BT Plaque-induced abnormalities in neurite geometry in transgenic models of Alzheimer disease: implications for neural system disruption.J Neuropathol Exp Neurol. 2001; 60: 753-758Crossref PubMed Scopus (83) Google Scholar which has also been noted in the APP23 mice, a model based on APP751 with APPsw mutation driven by the Thy-1 promoter.15Calhoun ME Wiederhold KH Abramowski D Phinney AL Probst A Sturchler-Pierrat C Staufenbiel M Sommer B Jucker M Neuron loss in APP transgenic mice.Nature. 1998; 395: 755-756Crossref PubMed Scopus (411) Google Scholar A problem that has plagued assessments of neuronal loss in brain regions that are also susceptible to amyloid deposition is the fact that amyloid deposits, especially in transgenic mice, tend to be dense structures that exclude other tissue elements from the central region of the deposit. They are, in essence, space-occupying lesions that, at the very least, displace neurons and other parenchymal elements that would normally occupy that volume. This certainly complicates the interpretation of neuronal counts in the APP23 model, even with unbiased stereologic methods.15Calhoun ME Wiederhold KH Abramowski D Phinney AL Probst A Sturchler-Pierrat C Staufenbiel M Sommer B Jucker M Neuron loss in APP transgenic mice.Nature. 1998; 395: 755-756Crossref PubMed Scopus (411) Google Scholar To address this issue, Schmitz and co-workers2Schmitz C Rutten BPF Pielen A Schäfer S Wirths O Tremp G Czech C Blanchard V Multhaup G Rezaie P Korr H Steinbusch HWM Pradier L Bayer TA Hippocampal neuronal loss exceeds amyloid plaque load in a transgenic mouse model of Alzheimer's disease.Am J Pathol. 2004; 164: 1495-1502Abstract Full Text Full Text PDF PubMed Scopus (221) Google Scholar performed an analysis in which they assumed similar density of neurons within the amyloid deposits as in the immediate surrounding tissue and mathematically added back the missing neurons in this area. With this “reconstructed” measurement, neuronal loss was not as marked, but still significantly different from controls. Thus, neuronal loss was not merely a counting artifact due to local perturbation of cell populations by dense amyloid deposits, which is a methodologic innovation that future quantitative studies of neuronal counts in mice with amyloid deposits should also consider implementing. In a recent report neuronal loss has been reported in the CA3 region of the hippocampus in older PDAPP mice,16Hsia AY Masliah E McConlogue L Yu GQ Tatsuno G Hu K Kholodenko D Malenka RC Nicoll RA Mucke L Plaque-independent disruption of neural circuits in Alzheimer's disease mouse models.Proc Natl Acad Sci USA. 1999; 96: 3228-3233Crossref PubMed Scopus (983) Google Scholar which, as mentioned above, was not detected in previous studies of neuronal integrity in PDAPP mice.13Irizarry MC Soriano F McNamara M Page KJ Schenk D Games D Hyman BT Aβ deposition is associated with neuropil changes, but not with overt neuronal loss in the human amyloid precursor protein V717F (PDAPP) transgenic mouse.J Neurosci. 1997; 17: 7053-7059Crossref PubMed Google Scholar The authors attributed the inconsistency to genetic background differences. The line with no neuronal loss was on an outbred background (C57BL/6 × DBA/2 × Swiss-Webster) whereas the line with hippocampal neuronal loss was maintained on a hybrid background (C57BL/6 × DBA/2). Another difference was the methodology used to count neurons. The negative study was based on unbiased stereology with Nissl-stained sections, while the study that showed neuronal loss used “stereologic counting principles” to count MAP-2 immunostained neurons with confocal microscopy. The possibility of a phenotype change (ie, loss of MAP-2 immunoreactivity), rather than actual neuronal loss, cannot be completely excluded with the latter method. An APP construct more similar to that used by Schmitz and co-workers2Schmitz C Rutten BPF Pielen A Schäfer S Wirths O Tremp G Czech C Blanchard V Multhaup G Rezaie P Korr H Steinbusch HWM Pradier L Bayer TA Hippocampal neuronal loss exceeds amyloid plaque load in a transgenic mouse model of Alzheimer's disease.Am J Pathol. 2004; 164: 1495-1502Abstract Full Text Full Text PDF PubMed Scopus (221) Google Scholar was used to generate the TgCRND8 mice as reported by Chishti and co-workers.17Chishti MA Yang DS Janus C Phinney AL Horne P Pearson J Strome R Zuker N Loukides J French J Turner S Lozza G Grilli M Kunicki S Morissette C Paquette J Gervais F Bergeron C Fraser PE Carlson GA George-Hyslop PS Westaway D Early-onset amyloid deposition and cognitive deficits in transgenic mice expressing a double mutant form of amyloid precursor protein 695.J Biol Chem. 2001; 276: 21562-21570Crossref PubMed Scopus (760) Google Scholar The transgene in TgCRND8 is APPsw and APPV717I on APP695, but under a different promoter, the hamster prion promoter (hPrP). This promoter has been successfully exploited in making a number of transgenic animal models;18Borchelt DR Davis J Fischer M Lee MK Slunt HH Ratovitsky T Regard J Copeland NG Jenkins NA Sisodia SS Price DL A vector for expressing foreign genes in the brains and hearts of transgenic mice.Genet Anal. 1996; 13: 159-163Crossref PubMed Scopus (297) Google Scholar it is widely expressed in the CNS and in peripheral tissues, but is not specific to neurons. Expression is high in the spinal cord, and spinal cord pathology complicates a number of animal models that use the PrP promoter. Nevertheless, the TgCRND8 mouse also has very early amyloid deposition (as early as 3 months) with neuritic dystrophy, reactive glial changes, and behavioral deficits. Evidence for neuronal loss in this model is lacking. At least three other bigenic PS1/APP transgenic models have been produced19Borchelt DR Ratovitski T van Lare J Lee MK Gonzales V Jenkins NA Copeland NG Price DL Sisodia SS Accelerated amyloid deposition in the brains of transgenic mice coexpressing mutant presenilin 1 and amyloid precursor proteins.Neuron. 1997; 19: 939-945Abstract Full Text Full Text PDF PubMed Scopus (880) Google Scholar, 20Holcomb L Gordon MN McGowan E Yu X Benkovic S Jantzen P Wright K Saad I Mueller R Morgan D Sanders S Zehr C O'Campo K Hardy J Prada CM Eckman C Younkin S Hsiao K Duff K Accelerated Alzheimer-type phenotype in transgenic mice carrying both mutant amyloid precursor protein and presenilin 1 transgenes.Nat Med. 1998; 4: 97-100Crossref PubMed Scopus (1145) Google Scholar, 21Dineley KT Xia X Bui D Sweatt JD Zheng H Accelerated plaque accumulation, associative learning deficits, and up-regulation of α 7 nicotinic receptor protein in transgenic mice co-expressing mutant human presenilin 1 and amyloid precursor proteins.J Biol Chem. 2002; 277: 22768-22780Crossref PubMed Scopus (200) Google Scholar (Table 1). As in the model reported by Schmitz and co-workers,2Schmitz C Rutten BPF Pielen A Schäfer S Wirths O Tremp G Czech C Blanchard V Multhaup G Rezaie P Korr H Steinbusch HWM Pradier L Bayer TA Hippocampal neuronal loss exceeds amyloid plaque load in a transgenic mouse model of Alzheimer's disease.Am J Pathol. 2004; 164: 1495-1502Abstract Full Text Full Text PDF PubMed Scopus (221) Google Scholar all have had early amyloid deposition, with neuritic dystrophy and reactive gliosis, as well as behavioral deficits. The latter have been the focus of a number of recent studies addressing the role of amyloid deposits on behavior.5Holcomb LA Gordon MN Jantzen P Hsiao K Duff K Morgan D Behavioral changes in transgenic mice expressing both amyloid precursor protein and presenilin-1 mutations: lack of association with amyloid deposits.Behav Genet. 1999; 29: 177-185Crossref PubMed Scopus (306) Google Scholar All three bigenic mice were generated by crossing APP transgenic mice with PS-1 transgenic mice, but no two are identical in terms of mutations, promoter, or background strain. In the other models APPsw, rather than a double APP mutation, is used and most use the PrP promoter rather than the Thy-1 promoter for both the APP and PS-1 parental transgenic mice. The choice of PS1 mutation also varies in the reported bigenic mice; however, the presence of the additional mutated PS-1 transgene in every case leads to accelerated amyloid deposition compared to the transgenic mice with only APP. Given that many of the transgenic mice reported to date have had robust amyloid pathology at relatively early ages, it would seem unlikely that the severity of amyloid deposition or time course of amyloid deposits is a major explanation for differences between the models.Table 1APP + PS-1 Bigenic Transgenic MiceAPP mutation (promoter)APP genetic backgroundPS-1 mutation/promoterPS-1 genetic backgroundReferenceAPP751sw+APPV717I (mo Thy-1)CBA× C57B6M146L (pHMG)C57B62Schmitz C Rutten BPF Pielen A Schäfer S Wirths O Tremp G Czech C Blanchard V Multhaup G Rezaie P Korr H Steinbusch HWM Pradier L Bayer TA Hippocampal neuronal loss exceeds amyloid plaque load in a transgenic mouse model of Alzheimer's disease.Am J Pathol. 2004; 164: 1495-1502Abstract Full Text Full Text PDF PubMed Scopus (221) Google ScholarAPP695sw (hPrP)C57B6/SJL× C57B6M146L (hPrP)SW/B6D2× B6D25Holcomb LA Gordon MN Jantzen P Hsiao K Duff K Morgan D Behavioral changes in transgenic mice expressing both amyloid precursor protein and presenilin-1 mutations: lack of association with amyloid deposits.Behav Genet. 1999; 29: 177-185Crossref PubMed Scopus (306) Google Scholar, 20Holcomb L Gordon MN McGowan E Yu X Benkovic S Jantzen P Wright K Saad I Mueller R Morgan D Sanders S Zehr C O'Campo K Hardy J Prada CM Eckman C Younkin S Hsiao K Duff K Accelerated Alzheimer-type phenotype in transgenic mice carrying both mutant amyloid precursor protein and presenilin 1 transgenes.Nat Med. 1998; 4: 97-100Crossref PubMed Scopus (1145) Google ScholarAPP695sw (hPrP)C57B6/SJLA246E (mo Thy-1)C57B6/SJL21Dineley KT Xia X Bui D Sweatt JD Zheng H Accelerated plaque accumulation, associative learning deficits, and up-regulation of α 7 nicotinic receptor protein in transgenic mice co-expressing mutant human presenilin 1 and amyloid precursor proteins.J Biol Chem. 2002; 277: 22768-22780Crossref PubMed Scopus (200) Google ScholarAPP695sw (hPrP)C3H/HeJ× C57B6A246E (hPrP)C3H/HeJ× C57B619Borchelt DR Ratovitski T van Lare J Lee MK Gonzales V Jenkins NA Copeland NG Price DL Sisodia SS Accelerated amyloid deposition in the brains of transgenic mice coexpressing mutant presenilin 1 and amyloid precursor proteins.Neuron. 1997; 19: 939-945Abstract Full Text Full Text PDF PubMed Scopus (880) Google Scholar, 27Wang J Tanila H Puolivali J Kadish I van Groen T Gender differences in the amount and deposition of amyloid β in APPswe and PS1 double transgenic mice.Neurobiol Dis. 2003; 14: 318-327Crossref PubMed Scopus (230) Google Scholarmo, mouse; h, hamster. Open table in a new tab mo, mouse; h, hamster. At present, neuronal loss has been assessed in only one of three bigenic animals, and this study failed to show neuronal loss, either globally in the cerebrum or in the hippocampal pyramidal cell layer.22Takeuchi A Irizarry MC Duff K Saido TC Hsiao Ashe K Hasegawa M Mann DM Hyman BT Iwatsubo T Age-related amyloid β deposition in transgenic mice overexpressing both Alzheimer mutant presenilin 1 and amyloid β precursor protein Swedish mutant is not associated with global neuronal loss.Am J Pathol. 2000; 157: 331-339Abstract Full Text Full Text PDF PubMed Scopus (206) Google Scholar Variance in the data precluded detection of neuronal loss less than 20%, but neuronal loss comparable to that reported by Schmitz and co-workers2Schmitz C Rutten BPF Pielen A Schäfer S Wirths O Tremp G Czech C Blanchard V Multhaup G Rezaie P Korr H Steinbusch HWM Pradier L Bayer TA Hippocampal neuronal loss exceeds amyloid plaque load in a transgenic mouse model of Alzheimer's disease.Am J Pathol. 2004; 164: 1495-1502Abstract Full Text Full Text PDF PubMed Scopus (221) Google Scholar (30 to 35%) should have been detectable. In considering the role of experimental variance in the differences between the two reports, it is worth noting that both studies were based on a relatively small number of bigenic animals (N = 422Takeuchi A Irizarry MC Duff K Saido TC Hsiao Ashe K Hasegawa M Mann DM Hyman BT Iwatsubo T Age-related amyloid β deposition in transgenic mice overexpressing both Alzheimer mutant presenilin 1 and amyloid β precursor protein Swedish mutant is not associated with global neuronal loss.Am J Pathol. 2000; 157: 331-339Abstract Full Text Full Text PDF PubMed Scopus (206) Google Scholar and N = 62Schmitz C Rutten BPF Pielen A Schäfer S Wirths O Tremp G Czech C Blanchard V Multhaup G Rezaie P Korr H Steinbusch HWM Pradier L Bayer TA Hippocampal neuronal loss exceeds amyloid plaque load in a transgenic mouse model of Alzheimer's disease.Am J Pathol. 2004; 164: 1495-1502Abstract Full Text Full Text PDF PubMed Scopus (221) Google Scholar). A control used by Takeuchi and co-workers22Takeuchi A Irizarry MC Duff K Saido TC Hsiao Ashe K Hasegawa M Mann DM Hyman BT Iwatsubo T Age-related amyloid β deposition in transgenic mice overexpressing both Alzheimer mutant presenilin 1 and amyloid β precursor protein Swedish mutant is not associated with global neuronal loss.Am J Pathol. 2000; 157: 331-339Abstract Full Text Full Text PDF PubMed Scopus (206) Google Scholar was APPsw littermates, while this control was not included in the Schmitz study2Schmitz C Rutten BPF Pielen A Schäfer S Wirths O Tremp G Czech C Blanchard V Multhaup G Rezaie P Korr H Steinbusch HWM Pradier L Bayer TA Hippocampal neuronal loss exceeds amyloid plaque load in a transgenic mouse model of Alzheimer's disease.Am J Pathol. 2004; 164: 1495-1502Abstract Full Text Full Text PDF PubMed Scopus (221) Google Scholar due to the breeding strategy. The Schmitz study2Schmitz C Rutten BPF Pielen A Schäfer S Wirths O Tremp G Czech C Blanchard V Multhaup G Rezaie P Korr H Steinbusch HWM Pradier L Bayer TA Hippocampal neuronal loss exceeds amyloid plaque load in a transgenic mouse model of Alzheimer's disease.Am J Pathol. 2004; 164: 1495-1502Abstract Full Text Full Text PDF PubMed Scopus (221) Google Scholar did use PS-1 littermate controls, however. A complicated issue to consider in assessing differences between transgenic models is the role genetic background might have had on the observed outcomes. This is an area of increasing research,23Ingram DK Jucker M Developing mouse models of aging: a consideration of strain differences in age-related behavioral and neural parameters.Neurobiol Aging. 1999; 20: 137-145Abstract Full Text Full Text PDF PubMed Scopus (62) Google Scholar but often relatively neglected. In some reports of transgenic animals, the genetic background information is not even included in the description; however, it is the consensus that this information should be mandatory in all reports of genetically modified mice.24Silva AJ Simpson EM Takahashi JS Lipp H-P Nakanishi S Wehner JM Giese KP Tully T Abel T Chapman PF Fox K Grant S Itohara S Lathe R Mayford M McNamara JO Morris RJ Picciotto M Roder J Shin H-S Slesinger PA Storm DR Stryker MP Tonegawa S Wang Y Wolfer DP Banbury Conference on Genetic Background in Mice: mutant mice and neuroscience: recommendation concerning genetic background.Neuron. 1997; 19: 755-759Abstract Full Text Full Text PDF PubMed Scopus (393) Google Scholar The role of genetic background certainly makes use of wild-type mice bred separately or purchased from the supplier suspect. It appears that this may have been the type of control chosen in the study by Schmitz and co-workers.2Schmitz C Rutten BPF Pielen A Schäfer S Wirths O Tremp G Czech C Blanchard V Multhaup G Rezaie P Korr H Steinbusch HWM Pradier L Bayer TA Hippocampal neuronal loss exceeds amyloid plaque load in a transgenic mouse model of Alzheimer's disease.Am J Pathol. 2004; 164: 1495-1502Abstract Full Text Full Text PDF PubMed Scopus (221) Google Scholar A better control would have been non-transgenic littermates; however, given that the mating was between homozygous PS-1 mice and hemizygous APPsw/APPV717I mice, a non-transgenic control was not possible. Littermate controls are preferable since they control for genetic background. The controls used by Takeuchi and co-workers22Takeuchi A Irizarry MC Duff K Saido TC Hsiao Ashe K Hasegawa M Mann DM Hyman BT Iwatsubo T Age-related amyloid β deposition in transgenic mice overexpressing both Alzheimer mutant presenilin 1 and amyloid β precursor protein Swedish mutant is not associated with global neuronal loss.Am J Pathol. 2000; 157: 331-339Abstract Full Text Full Text PDF PubMed Scopus (206) Google Scholar included non-transgenic littermates. Despite this concern, it is unlikely that use of sub-optimal controls is the explanation for differences between the two studies; however, it is worth noting that the most robust statistical differences were between the wild-type controls and the bigenic mice rather than between the PS-1 littermates and the bigenic mice.2Schmitz C Rutten BPF Pielen A Schäfer S Wirths O Tremp G Czech C Blanchard V Multhaup G Rezaie P Korr H Steinbusch HWM Pradier L Bayer TA Hippocampal neuronal loss exceeds amyloid plaque load in a transgenic mouse model of Alzheimer's disease.Am J Pathol. 2004; 164: 1495-1502Abstract Full Text Full Text PDF PubMed Scopus (221) Google Scholar The effect of genetic background is a recognized factor in determining complex behavioral and neuronal phenotypes in genetically modified mice.23Ingram DK Jucker M Developing mouse models of aging: a consideration of strain differences in age-related behavioral and neural parameters.Neurobiol Aging. 1999; 20: 137-145Abstract Full Text Full Text PDF PubMed Scopus (62) Google Scholar Since there is no “normal” mouse strain, it is essential to control for genetic background using strategies that have been recommended by a recent consensus committee, including back-crossing onto standard laboratory strains, such as C57B6.24Silva AJ Simpson EM Takahashi JS Lipp H-P Nakanishi S Wehner JM Giese KP Tully T Abel T Chapman PF Fox K Grant S Itohara S Lathe R Mayford M McNamara JO Morris RJ Picciotto M Roder J Shin H-S Slesinger PA Storm DR Stryker MP Tonegawa S Wang Y Wolfer DP Banbury Conference on Genetic Background in Mice: mutant mice and neuroscience: recommendation concerning genetic background.Neuron. 1997; 19: 755-759Abstract Full Text Full Text PDF PubMed Scopus (393) Google Scholar An example of the importance of genetic background in transgenic models of AD is the recent report that behavioral deficits observed in outbred APPsw transgenic animals were not detected when the animals were back-crossed to produce congenic mice.25Savonenko AV Xu GM Price DL Borchelt DR Markowska AL Normal cognitive behavior in two distinct congenic lines of transgenic mice hyperexpressing mutant APP SWE.Neurobiol Dis. 2003; 12: 194-211Crossref PubMed Scopus (71) Google Scholar Another caveat of the study by Schmitz and co-workers2Schmitz C Rutten BPF Pielen A Schäfer S Wirths O Tremp G Czech C Blanchard V Multhaup G Rezaie P Korr H Steinbusch HWM Pradier L Bayer TA Hippocampal neuronal loss exceeds amyloid plaque load in a transgenic mouse model of Alzheimer's disease.Am J Pathol. 2004; 164: 1495-1502Abstract Full Text Full Text PDF PubMed Scopus (221) Google Scholar was that the analysis was restricted to female animals. The explanation for limiting the analysis to females is not given, but sex has been shown to influence the phenotype of several transgenic models of AD, most notably the descriptions of more severe amyloid deposition in female APP mice26Callahan MJ Lipinski WJ Bian F Durham RA Pack A Walker LC Augmented senile plaque load in aged female β-amyloid precursor protein-transgenic mice.Am J Pathol. 2001; 158: 1173-1177Abstract Full Text Full Text PDF PubMed Scopus (230) Google Scholar, 27Wang J Tanila H Puolivali J Kadish I van Groen T Gender differences in the amount and deposition of amyloid β in APPswe and PS1 double transgenic mice.Neurobiol Dis. 2003; 14: 318-327Crossref PubMed Scopus (230) Google Scholar and more marked neurofibrillary pathology in female tau mice.28Lewis J McGowan E Rockwood J Melrose H Nacharaju P Van Sleghtenhorst M Gwinn-Hardy K Murphy MP Baker M Yu X Duff K Hardy J Corral A Lin W Yen SH Dickson DW Davies P Hutton M Neurofibrillary tangles, amyotrophy, and progressive motor disturbance in tau mutant (P301L) transgenic mice.Nat Genet. 2000; 25: 402-405Crossref PubMed Scopus (1113) Google Scholar For some transgenic mice, most notably Tg2576,27Wang J Tanila H Puolivali J Kadish I van Groen T Gender differences in the amount and deposition of amyloid β in APPswe and PS1 double transgenic mice.Neurobiol Dis. 2003; 14: 318-327Crossref PubMed Scopus (230) Google Scholar male mice are unusually aggressive and need to be housed singly. This creates yet another difference when considering behavioral and pathological phenotypic differences between males and females, since the nature of housing has been shown to influence the phenotype. For example, Jankowsky and co-workers recently showed that environmental enrichment is associated with a greater amyloid burden in APP transgenic mice.29Jankowsky JL Xu G Fromholt D Gonzales V Borchelt DR Environmental enrichment exacerbates amyloid plaque formation in a transgenic mouse model of Alzheimer disease.J Neuropathol Exp Neurol. 2003; 62: 1220-1227Crossref PubMed Scopus (167) Google Scholar In summary, the model described by Schmitz and co-workers2Schmitz C Rutten BPF Pielen A Schäfer S Wirths O Tremp G Czech C Blanchard V Multhaup G Rezaie P Korr H Steinbusch HWM Pradier L Bayer TA Hippocampal neuronal loss exceeds amyloid plaque load in a transgenic mouse model of Alzheimer's disease.Am J Pathol. 2004; 164: 1495-1502Abstract Full Text Full Text PDF PubMed Scopus (221) Google Scholar has significant value as a model for neuronal loss associated with amyloid deposition if the results can be confirmed in a larger series of animals with appropriate non-transgenic littermate controls and if the neuronal loss can be shown to be anatomically relevant to that found in AD. The temporal sequence of biochemical and cellular pathology, as well as the behavioral correlates of amyloid-associated neuronal loss are important outcome variables that remain to be determined.