A Novel Mouse Model of Alzheimer's Disease with Chronic Estrogen Deficiency Leads to Glial Cell Activation and Hypertrophy

Figure 1

Generation of chimeric mice. A construct used to generate chimeric mice expressing human APP with Swedish mutation and hypoestrogenism due to the absence of FSH receptor. A murine prion protein (PrP) genomic fragment was modified to obtain the vector phgPrP [21]. A region containing 6 kb of promoter, exon 1 (noncoding), intron 1, and exon 2 (non-coding) was fused to the third and last exon (2008 bp) and 2.2 kb of downstream sequence. The subsequent removal of the PrP open reading frame (ORF) generated the vector MoPrP.Xho, in which the cDNA encoding PS1 lacking the exon 9 (amino acids 290–319) was subcloned [44]. Finally, the region encoding residues 592 to 622 from the mouse APP-695 cDNA was substituted by the corresponding human one (hatched area), which contains the Swedish KM to NL double mutation and the Aβ42 coding sequence. The resulting hybrid mouse/human cDNA was subcloned in MoPrP.Xho [11]. Half arrows indicate the position of the primers used for genotyping the mice. For complete inactivation of the FSH receptor gene, a 638 bp fragment containing part of the 5′ untranslated region, the coding region of exon 1 (101 bp), and about 129 bp of intron 1 was substituted by a 1.7 kb neomycin expression cassette (dotted box) [19]. A multiplex PCR using primers indicated by half arrows was used to genotype the offspring [16]. Black boxes: non-coding PrP exons 1 and 2; grey box: coding exon; ORF are boxed.