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. 2017 Jan 19;12(1):e0170450.
doi: 10.1371/journal.pone.0170450. eCollection 2017.

Perinatal Choline Supplementation Reduces Amyloidosis and Increases Choline Acetyltransferase Expression in the Hippocampus of the APPswePS1dE9 Alzheimer's Disease Model Mice

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Perinatal Choline Supplementation Reduces Amyloidosis and Increases Choline Acetyltransferase Expression in the Hippocampus of the APPswePS1dE9 Alzheimer's Disease Model Mice

Tiffany J Mellott et al. PLoS One. .
Free PMC article

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Abstract

Prevention of Alzheimer's disease (AD) is a major goal of biomedical sciences. In previous studies we showed that high intake of the essential nutrient, choline, during gestation prevented age-related memory decline in a rat model. In this study we investigated the effects of a similar treatment on AD-related phenotypes in a mouse model of AD. We crossed wild type (WT) female mice with hemizygous APPswe/PS1dE9 (APP.PS1) AD model male mice and maintained the pregnant and lactating dams on a control AIN76A diet containing 1.1 g/kg of choline or a choline-supplemented (5 g/kg) diet. After weaning all offspring consumed the control diet. As compared to APP.PS1 mice reared on the control diet, the hippocampus of the perinatally choline-supplemented APP.PS1 mice exhibited: 1) altered levels of amyloid precursor protein (APP) metabolites-specifically elevated amounts of β-C-terminal fragment (β-CTF) and reduced levels of solubilized amyloid Aβ40 and Aβ42 peptides; 2) reduced number and total area of amyloid plaques; 3) preserved levels of choline acetyltransferase protein (CHAT) and insulin-like growth factor II (IGF2) and 4) absence of astrogliosis. The data suggest that dietary supplementation of choline during fetal development and early postnatal life may constitute a preventive strategy for AD.

Conflict of interest statement

The authors have declared that no competing interests exist.

Figures

Fig 1
Fig 1. Aβ levels in the hippocampus of APP.PS1 mice.
Hippocampal lysates from females and males were used to measure solubilized Aβ40 (A, B) and Aβ42 (C, D) levels by ELISA, and APP and soluble Aβ (E, F) levels by Western blot analysis using the anti-APP 6E10 antibody. For each sex, the lysates from both 9- and 12-month-old mice were loaded on the same SDS-page gel and immunoblotted together, and therefore, the data were analyzed together and presented as percentages of the 9-month control values. As determined by 2-way ANOVA for genotype and diet and Tukey test per age: * represents p<0.05 compared to control diet APP.PS1 mice at 9-months; †, p<0.05 compared to choline-supplemented diet APP.PS1 mice at 9-months; and #, p<0.05 compared to control diet APP.PS1 mice at the same age.
Fig 2
Fig 2. APP metabolite levels in the hippocampus of APP.PS1 mice.
Hippocampal lysates from 12-month-old female and male APP.PS1 mice were used to measure the α- and β-CTFs using an anti-C-terminal APP antibody. As determined by Student T-test, * represents p<0.05 compared to control diet APP.PS1 mice of the same sex.
Fig 3
Fig 3. Aβ40 plaques in the hippocampus of 9-, and 12-month old WT and APP.PS1 mice.
Anterior (A) and posterior (B) hippocampal sections from representative 9-month females stained with anti-Aβ40. The average number of Aβ40 plaques per animal (C, D) and the total Aβ40 plaque area (E, F) were quantified using ImageJ64 software in both females and males. As determined by 2-way ANOVA for hippocampal region and diet and Tukey test per age, # represents p<0.05 compared to control diet APP.PS1 mice at the same age. There was a significant overall effect of choline supplementation on the average number and total plaque area in both females (P270: average number p<0.001 and total plaque area p<0.0005; P360: p<0.0001 and p<0.0001) and males (P270: p<0.05 and p<0.05; P360: p<0.0005 and p<0.01).
Fig 4
Fig 4. Aβ42 plaques in the hippocampus of 9-, and 12-month old WT and APP.PS1 mice.
Anterior (A) and posterior (B) hippocampal sections from representative 9-month females stained with anti-Aβ42. The average number of Aβ42 plaques per animal (C, D) and the total Aβ42 plaque area (E, F) were quantified using ImageJ64 software in both females and males. As determined by 2-way ANOVA for hippocampal region and diet and Tukey test per age, # represents p<0.05 compared to control diet APP.PS1 mice at the same age. There was a significant overall effect of choline supplementation on the average number and total plaque area in both females (P270: average number p<0.005 and total plaque area p<0.01; P360: p<0.01 and p<0.05) and males (P270: p<0.05 and p<0.05; P360: p<0.005 and p<0.0005).
Fig 5
Fig 5. CHAT protein levels in the hippocampus of 9- and 12-month old WT and APP.PS1 mice.
Hippocampal lysates were used to measure CHAT protein levels by Western blot analysis in females (A) and males (B). As determined by 2-way ANOVA for genotype and diet and Tukey test per age: * represents p<0.05 compared to control diet WT mice at the same age; and #, p<0.05 compared to control diet APP.PS1 mice at the same age.
Fig 6
Fig 6. Neurogenesis in the hippocampus WT and APP.PS1 mice.
DCX immunofluorescence staining of anterior hippocampal sections (A) from 9-month old female mice visualized using confocal microscopy. Bar represents 50 μm. Hippocampal lysates of 9- and 12-month old females (B) and males (C) were used to measure DCX protein levels by Western blot analysis. As determined by 2-way ANOVA for genotype and diet and Tukey test per age: * represents p<0.05 compared to control diet WT mice at the same age; and #, p<0.05 compared to control diet APP.PS1 mice at the same age. There was a significant overall effect of perinatal choline supplementation on DCX protein levels in both 9- and 12-month females (p<0.05 and p<0.0005, respectively) and males (p<0.001 and p<0.01, respectively).
Fig 7
Fig 7. Astrogliosis levels in the hippocampus WT and APP.PS1 mice.
GFAP immunofluorescence staining of anterior hippocampal sections (A) from 9-month old female mice visualized using confocal microscopy. Bar represents 50 μm. Hippocampal lysates of 9- and 12-month old females (B) and males (C) were used to measure GFAP protein levels by Western blot analysis. As determined by 2-way ANOVA for genotype and diet and Tukey test per age: * represents p<0.05 compared to control diet WT mice at the same age; †, p<0.05 compared to choline-supplemented diet WT mice at the same age; and #, p<0.05 compared to control diet APP.PS1 mice at the same age.
Fig 8
Fig 8. Hippocampal IGF2 protein levels in WT and APP.PS1 mice.
Hippocampal lysates of 9-old females and males were used to measure IGF2 protein levels by Western blot analysis. In the males, there was a significant effect of choline supplementation, regardless of genotype, using a 2-way ANOVA (p<0.005). As determined by 2-way ANOVA for genotype and diet and Tukey test per age: * represents p<0.05 compared to control diet WT mice of the same sex; and #, p<0.05 compared to control diet APP.PS1 mice of the same sex.

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