Systematic phenotyping and characterization of the 5xFAD mouse model of Alzheimer’s disease.
Open accessAbstract
Mouse models of human diseases are invaluable tools for studying pathogenic mechanisms and testing interventions and therapeutics. For disorders such as Alzheimer's disease in which numerous models are being generated, a challenging first step is to identify the most appropriate model and age to effectively evaluate new therapeutic approaches. Here we conducted a detailed phenotypic characterization of the 5xFAD model on a congenic C57BL/6 J strain background, across its lifespan - including a seldomly analyzed 18-month old time point to provide temporally correlated phenotyping of this model and a template for characterization of new models of LOAD as they are generated. This comprehensive analysis included quantification of plaque burden, Aβ biochemical levels, and neuropathology, neurophysiological measurements and behavioral and cognitive assessments, and evaluation of microglia, astrocytes, and neurons. Analysis of transcriptional changes was conducted using bulk-tissue generated RNA-seq data from microdissected cortices and hippocampi as a function of aging, which can be explored at the MODEL-AD Explorer and AD Knowledge Portal. This deep-phenotyping pipeline identified novel aspects of age-related pathology in the 5xFAD model.
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Female microglia exhibit heightened plaque pathology compared to age-matched male microglia in the 5xFAD model.
“female microglia exhibited heightened plaque pathology compared to age-matched male microglia in the 5xFAD model.70,71”
From Amyloid-β induces lipid droplet-mediated microglial dysfunction via the enzyme DGAT2 in Alzheimer’s disease (2025) · cites this paper as [71]
In Technical Validation (Fig. 4), the paper reports more cortical plaques in female than male 5xFAD mice at 4, 12, and 18 months. Its microgliosis section (Fig. 6) says female mice tend to have higher microglial densities, but it does not establish that female microglia themselves exhibit heightened plaque pathology. (LLM, conf=0.86)
Experimental — accuracy not yet measuredThe possibility that rostral-to-caudal effects are downstream of regional differences in promoter activity can probably be ruled out because many teams have shown strong expression of the transgene and high levels of amyloid plaque deposition in thalamus and midbrain.
“This raises the possibility that rostral-to-caudal effects we have detected are downstream of regional differences in promoter activity. This can probably be ruled out because many teams have shown strong expression of the transgene and high levels of amyloid plaque deposition in thalamus and midbrain81—regions in which we have demonstrated highly significant reductions in volume (Ref20, their”
From Magnetic resonance microscopy maps widespread effects of Alzheimer’s disease on brain structures and behavior in mice (2026) · cites this paper as [20]
Split into 2 atoms — 0 confirmed, 2 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.
Experimental — accuracy not yet measuredAmyloid pathology is more accelerated and consistent in female 5xFAD mice compared with male 5xFAD mice.
“amyloid pathology is more accelerated and consistent in female 5xFAD mice compared with male 5xFAD mice (Forner et al., 2021; Sasmita et al., 2025).”
From Alzheimer’s genetic risk factor CASS4 controls microglial focal adhesion biology and amyloidosis (2026) · cites this paper as Forner et al., 2021
The Usage Notes state that female 5xFAD mice develop pathology before males, and Technical Validation reports higher cortical plaque numbers in females at several ages. The Usage Notes call plaque pathology reproducible overall, but do not say it is more consistent in females than in males. No page number is available in our copy. (LLM, conf=0.88)
Experimental — accuracy not yet measuredOne representative brain slice from each mouse within identical experimental cohort was subjected to simultaneous staining in the same container.
“One representative brain slice from each mouse within the identical experimental cohort (comprising the same genotype, age, and sex) was subjected to simultaneous staining in the same container as described [25, 27].”
From APOE Christchurch enhances a disease-associated microglial response to plaque but suppresses response to tau pathology (2025) · cites this paper as [25]
The Methods section on Imaris quantitative analysis says comparable sections from each animal were processed at the same time using the same batches of solutions. It does not say that exactly one representative slice per mouse was stained, that mice were grouped by identical genotype, age, and sex, or that slices shared a container. Page numbers are unavailable in our copy. (LLM, conf=0.85)
Experimental — accuracy not yet measuredMice from both sexes were used unless otherwise specified; when both male and female mice were used, an analysis for sex differences was performed; unless otherwise noted, no sex differences were observed.
“Given the reported sex differences in some disease pathology phenotypes46,47,62, mice from both sexes were used unless otherwise specified. When both male and female mice were used, an analysis for sex differences was performed; unless otherwise noted, no sex differences were observed.”
From Lymphoid gene expression supports neuroprotective microglia function (2025) · cites this paper as [46]
The 5xFAD mouse expresses human APP with three FAD mutations and human PSEN1 with two FAD mutations, and is commonly used to study the mechanisms of AD.
“The transgenic mouse model, 5xFAD, expresses human APP with three FAD mutations and human PSEN1 with two FAD mutations, and is commonly used to study the mechanisms of AD (Oakley et al., 2006; Forner et al., 2021).”
From Age-dependent microstructure alterations in 5xFAD mice by high-resolution diffusion tensor imaging (2022) · cites this paper as Forner et al., 2021
Sample sizes were not predetermined using statistical methods.
“The sample sizes are similar to those found in prior studies conducted by MODEL-AD and were not predetermined using statistical methods [25].”
From APOE Christchurch enhances a disease-associated microglial response to plaque but suppresses response to tau pathology (2025) · cites this paper as [25]
The 5xFAD mouse model has been reported to exhibit sex-specific differences in phenotype.
“As the 5xFAD mouse model has been reported to exhibit sex-specific differences in phenotype, data separated by sex of significant differences are included in Supp. Figure 3, 4i-x, 6 g-l [25, 49].”
From APOE Christchurch enhances a disease-associated microglial response to plaque but suppresses response to tau pathology (2025) · cites this paper as [25]
All data were analyzed by two-way ANOVA to adjust for sex differences.
“All data were analyzed by two-way analysis of variance (ANOVA) to adjust for sex differences given prior reports of sex-dependent effects on pathology in the 5xFAD model [13], with a Tukey’s post-hoc test using GraphPad Prism 10.”
From Astrocytic APOE3-Christchurch expression ameliorates brain amyloid-β pathology in 5xFAD mice (2026) · cites this paper as [13]
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- Authors
- Forner, S., Kawauchi, S., Balderrama-Gutierrez, G., Kramár, E. A., Matheos, D. P., Phan, J., Javonillo, D. I., Tran, K. M., Hingco, E., da Cunha, C., Rezaie, N., Alcantara, J. A., Baglietto-Vargas, D., Jansen, C., Neumann, J., Wood, M. A., MacGregor, G. R., Mortazavi, A., Tenner, A. J., LaFerla, F. M., & Green, K. N.
- Affiliations
- University of California, Irvine (US)
- Corresponding author
- Green, Kim N.
- ORCID
- Venue
- Scientific Data
- Published
- 2021-10-15
- Volume
- 8
- Issue
- 1
- Pages
- 270-270
- ISSN
- 2052-4463 (ISSN-L)
- PMC
- PMC8519958
- OpenAlex
- W3207056385
- Cited by
- 501
- References
- 41
- Citations by year
- 2026: 120 · 2025: 144 · 2024: 96 · 2023: 90 · 2022: 49 · 2021: 2
- Type
- data paper
- Language
- en
- Open access
- gold
- doi.org
- Locations
- Scientific Data (publishedVersion, cc-by)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
- Europe PMC (PubMed Central) (publishedVersion, other-oa)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- eScholarship (California Digital Library) (submittedVersion, cc-by)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- University of California, Irvine
- National Institute on Aging (AG054345, RF1AG055104, U54 AG054349 S1-9, AG055104, RF1AG056768, U54AG054345, U54AG054349)
- Topics
- Health Sciences › Medicine › Physiology › Alzheimer's disease research and treatments
- Health Sciences › Medicine › Psychiatry and Mental health › Dementia and Cognitive Impairment Research
- Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
- Keywords
- 5xFAD mice, Alzheimer's disease, amyloid-beta, amyloid plaques, neuropathology, cognitive assessment, behavioral assessment, microglia, astrocytes, neurons, transcriptomics, aging, C57BL/6J mice
- MeSH terms
- RNA-Seq, Alzheimer Disease, Animals, Behavior, Animal, Disease Models, Animal, Hippocampus, Mice, Inbred C57BL, Synaptic Transmission, Phenotype, Long-Term Potentiation, Mice
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