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5XFAD

5XFAD

 Poccessing five transgenic mutation, 5XFAD have drastically different gene expression (1300 differentially expressed gene at 4 months) compared to wild type. Three mutation are on APP and two mutations are on PS1, together they grant 5XFAD huge production rate of Aβ42, which is a highly amyloidogenic and thus can lead to severe amyloid pathology in various regions in CNS, ranging from subiculum to cortical layer V, from hippocampus to thalamus, etc. 

   Amyloid pathology in 5XFAD is associated with astrogliosis and microgliosis, and the later is in turn linked with microvascular damage, which is another angiopathy linked with amyloidosis beside the progressive cerebral amyloid angiopathy (CAA).

   Worse still, amyloidosis in 5XFAD is also linked with neurites dystrophy, ranging from axonal abnormality to progressive myelin abnormalities, which might be the underlying cause for the synaptic dysfunction in 5XFAD, ranging from decrement in excitability to even synapse loss.

   The synaptic dysfunction might in turn be the reason for the neurodegeneration, especially in cortical layer V, which experience up to about 40% neuronal loss.

   Reasonably, the severe neurodegeneration in 5XFAD might be the cause of its cognitive dysfunction, including progressive working memory deficits and decrement in anxiety, and behavioral dysfunction, including sensorimotor impairments and aberrant reflexes.

   Tauopathy, on the other hand, is not known in 5XFAD.

   Therefore, all in all, 5XFAD might be a great AD model to isolate its amyloid pathology out, amplify and then investigate into its effects within a short time span

Add Animal Model Information


Genetics

  • Familiar Alzheimer’s Disease (FAD) Mutation:
    -On human Amyloid beta (A4) Precursor Protein (APP)(695) (mouse/human chimera)
    Swedish(K670N, M671L)
    London(V717I)
    Florida(I716V)
    -On Presenilin1 (human)
    M146L (A>C)
    L286V
    (The Jackson Laboratory)
    (ALZFORUM)
  • Regulation of Mutation Overexpression: Neural-specific elements in mouse Thy1 promoter(The Jackson Laboratory)
  • Mouse hemizygous for the transgene are fertile and viable,(The Jackson Laboratory)
  • While homozygous animal show more serious amyloid pathology and behavioral dysfunction, which also onset earlier.(ALZFORUM)
  • The pathological phenotype of this strain is less robust than its cogenic transgenic model C57BL/6(The Jackson Laboratory)
  • Yet, the retinal degeneration allele Pde6brd1 is absent in this strain(The Jackson Laboratory)
  • Suggested Control (Genetic Background): Noncarrier C57BL/6J(The Jackson Laboratory)

Amyloid Pathology

  • This strain possess:
    Three mutations on APP, which can increase the APP cleavage by β-secretase and thus increase the production of Aβ40 and Aβ42.
    Two mutations on PS1, which is a component of γ-secretase and thus can affect its cleavage activity to increase the production ratio of Aβ42(a more toxic form) to Aβ40 (5XFAD generate Aβ almost exclusively in the form of Aβ42).(The Jackson Laboratory)
  • Therefore, 5XFAD readily recapitulate amyloid pathology of AD (homozygous is even more severe than hemizygous) and thus is potentially a suitable AD model for amyloid plaque formation and intraneuronal Aβ-42 induced neurodegeneration(The Jackson Laboratory)
  • Subiculum and Cortical layer V are regions having the amyloid pathology with the earliest onset and highest severity.(ALZFORUM)
  • Subicular and layer V pyramidal neurons contain detectable (with immunohistochemistry) human APP as early as p16, and observable intraneuronal Aβ as early as 6 weeks.(ALZFORUM)
  • Cortex, hippocampus and thalamus contain detectable(with immunohistochemistry) extracellular amyloid plaques as early as 2-months.(ALZFORUM)
  • Spinal cord contain observable extracellular plaques and intraneuronal Aβ by 3 months.(ALZFORUM)
  • Frontal, entorhinal, parietal cortices and dentate gyrus exhibit Thioflavin-S-positive plaques between 2 and 4 months.(ALZFORUM)
  • Superficial, leptomeningeal vessel exhibit progressive cerebral amyloid angiopathy (CAA) start from about 3 months.(ALZFORUM)
  • Progressive microcapillary leakage is more serious in transgenic animals,(ALZFORUM)
  • And microvascular damage is linked with microgliosis,(ALZFORUM)
  • And microgliosis, together with astrogliosis and dystrophic neurites are linked with amyloid plaques in this model.(ALZFORUM)
  • Thus, amyloid pathology might have some association with both angiopathy.(ALZFORUM)
  • (although the amyloid rarely deposit abut microvessels.)(ALZFORUM)

Neurodegeneration

  • This strain might potentially be a suitable AD model for Aβ-42 induced neurodegeneration.(The Jackson Laboratory)
  • Layer V pyramidal neurons was observed to experience roughly 40% loss in one year-old mice.(ALZFORUM)
  • Hippocampal CA1 and cortex overall, nonetheless, do not show significant difference from wild-type mice in term of neuronal population size.(ALZFORUM)

Synaptic Dysfunction

  • The synaptic deficits in 5XFAD can be evident electrophysiologically from 2 to 3 months.(ALZFORUM)
  • 5XFAD mice exhibit decrement in the level of synaptic marker protein(The Jackson Laboratory)
  • Loss of specific synapses are also observable in 5XFAD mice by crossing them with mice expressing Yellow Fluorescent Protein(YFP), and thereby generating the 5xFAD-YFP strain.(ALZFORUM)
  • Pyramidal neurons in prefrontal and somatosensory cortices, for instance, exhibit decrement in spine density at 6 months.(ALZFORUM)
  • Pyramidal neurons in hippocampi, nonetheless, does not exhibit this decrement.(ALZFORUM)
  • Layer V neurons exhibit decrement in frequencies and amplitudes of miniature excitatory postsynaptic currents, which infer pre- and post-synaptic dysfunction, respectively, which might be the cause of their lower excitability.(ALZFORUM)
  • The lower excitability of layer V neruons might in turn be the cause of their abnormality in synaptic plasticity as well. For instance, the stimulation protocol, which induce spike-timing-dependent Long Term Potentiation in wild-type, induce Long Term Depression in 5XFAD instead.(ALZFORUM)
  • Layers II/III neurons, nevertheless, do not have different property between genotypes.(ALZFORUM)
  • Region-specific and age-dependent differences in neural activity are observed between 5XFAD and wild type with manganese-enhanced magnetic resonance imaging (MEMRI) regional activity mapping, for instance,(ALZFORUM)
  • Activity in amygdala and hippocampus are consistently increased in 5XFAD between 1 and 5 months or perhaps even longer.(ALZFORUM)

Neurite Dystrophy

  • Progressive myelin abnormalities onset as early as 1 month(ALZFORUM)
  • Thinner myelin sheaths and decrement in myelin basic protein level are observed in various brain regions of 5XFAD at 1 month.(ALZFORUM)
  • Progressive decrement in the thickness of myelin and other myelin morphological abnormality, ranging from myelin outfolding to ballooned myelin, from split myelin to double myelin sheaths on axons, have higher frequence and earlier onset in 5XFAD(ALZFORUM)
  • Axon calibers drop in retrosplenial cortex and CA1 at 2 to 3 months(ALZFORUM)
  • Axonal swellings, which is an indicator of axon degeneration, are observed in various brain regions as early as 3 months, but it is independent of the plaques.(ALZFORUM)

Cognitive Deficits

  • Memory impairment in Y-maze test is exhibited.(The Jackson Laboratory)
  • Progressive working memory deficits (demonstrated in elevated plus-maze) and progressive decrement in anxiety (evaluated via elevated plus-maze) onset between 3 and 6 months(ALZFORUM)
  • Decrement in anxiety is observed in open field, but differences between 5XFAD and wild type are not observed until 9 to 12 months(ALZFORUM)
  • In Morris water maze test, 5XFAD show clear preference(more than 25% retention time) for the target quadrant in a probe test as normal at least until 7 months. Nonetheless, their retention time is still a bit less compared to wild type.(ALZFORUM)
  • There is also statistically significant, although small, difference between 5XFAD and wild type in term of the number of platform crossings.(ALZFORUM)

Behavioral Dysfunction

  • Compared to hemizygous, homozygous 5XFAD have higher severity and earlier onset of behavioral deficits(ALZFORUM)
  • Exploratory behavior, measured by the arm entries number, does not differ much between 5XFAD and wild type at least until 1 year.(ALZFORUM)
  • Sensorimotor impairments in string-suspension and balance-beam are observable at 9 months(ALZFORUM)
  • Aberrant reflexes, particularly hind and forelimb clasping in tail-suspension test, are observable as early as 5 months(ALZFORUM)

Genomics

  • Number of differentially expressed gene between 5XFAD and wild type in hippocampi increase drastically from 42 transcripts at 2 months to over 1300 at 4 months.(ALZFORUM)
  • Those upregulated genes in 4 months show an enrichment for terms linked with immune activation in gene ontology analysis.(ALZFORUM)
  • The number of differentially expressed gene exhibits female-biased sexual dimorphism.(ALZFORUM)
  • For instance, amidst the 5XFAD mice at 4 months, females have slightly higher levels of human APP and PSEN1 mRNAs.(ALZFORUM)

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