Abstract
Amyloid-β (Aβ) plays a critical role as a biomarker in Alzheimer’s disease (AD) diagnosis. In addition to its diagnostic potential in the brain, recent studies have suggested that changes of Aβ level in the plasma can possibly indicate AD onset. In this study, we found that plasma Aβ(1–42) concentration increases with age, while the concentration of Aβ(1–42) in the cerebrospinal fluid (CSF) decreases in APPswe, PS1M146V and TauP301L transgenic (3xTg-AD) mice, if measurements were made before formation of ThS-positive plaques in the brain. Our data suggests that there is an inverse correlations between the plasma and CSF Aβ(1–42) levels until plaques form in transgenic mice’s brains and that the plasma Aβ concentration possesses the diagnostic potential as a biomarker for diagnosis of early AD stages.
Similar content being viewed by others
Introduction
The diagnostic potential of plasma amyloid-β (Aβ) in Alzheimer’s disease (AD) has been receiving attention because previous clinical studies indicated a possible relationship between increased risk of AD and lowered Aβ(1–42)/Aβ(1–40) ratio due to the decrease in plasma Aβ(1–42) concentrations1,2,3,4,5. A meta-analysis of 13 studies that assessed the potential of plasma Aβ as a diagnostic tool also reported a possibility of increased plasma Aβ(1–40) leading to subsequent cognitive decline6. If the Aβ concentration changes in the plasma can reflect the progression of AD in patients, it would enable the diagnosis of AD using less costly and less invasive methods. However, because there are studies that found either decreased or no meaningful changes in the plasma Aβ level of AD patients7,8,9, the potential of using the plasma Aβ measurements for AD diagnosis has been continuously debated. At this point, there is not enough evidence to acknowledge the level of plasma Aβ as a reliable biomarker for AD diagnosis yet.
When human AD studies build their AD patient pool, the diagnosis of probable AD is made based on the patient’s cognitive impairments described by the criteria of the National Institute of Neurological Disorders and Stroke and the AD and Related Disorders Association10,11. These human studies select patients with clinical diagnosis and study their Aβ abnormalities because the Aβ plaque deposition proceeds cognitive impairments in AD patients12,13. Current AD diagnosis can be confirmed with Aβ abnormalities detected by neuroimaging. Alternatively, measurements of Aβ in the cerebrospinal fluid (CSF) is known to reflect AD pathology in the brain14,15,16. If there were a method to diagnose AD prior to the Aβ plaque deposition in the brain, diagnosis and treatment of patients in early AD stages would be possible, even before at risk individuals develop cognitive impairments. This possibility led us to search for plasma biomarkers with diagnostic potentials for early AD stage.
A significant correlation between Aβ concentrations in the CSF and plasma were identified when we measured the Aβ levels prior to the plaque formation in the brain of transgenic mice. However, such correlations gradually disappeared as our mice aged and developed Aβ plaques in their brains. We hypothesized that the concentration changes in plasma Aβ can be utilized as a reliable biomarker for early diagnosis of AD prior to the plaque formation. In order to evaluate our hypothesis, we selected APPswe, PS1M146V and TauP301L transgenic (3xTg-AD) mice, which imitate human AD pathophysiologies including age-dependent behavioral and cognitive alterations. The 3xTg-AD mice show cognitive deficits in the Morris water maze at 6 months of age and present visible Aβ plaque deposition in the brain at 6 months of age17,18,19,20,21. They are also known to develop Aβ plaques more slowly compared with other transgenic mice, making the 3xTg-AD model fit for studying the relationship between AD and the plasma and CSF Aβ level alterations before the plaque formation.
In this study, we first confirmed the absence of ThS positive amyloid plaques, dense-core plaques, in the brains of young 3xTG-AD mice with immunohistochemical staining. We then measured the changes in concentration of Aβ(1–42) and Aβ(1–40) in the CSF and plasma of 5-, 7-, 9- and 12-month-old female transgenic mice using sandwich-ELISA to define a relationship between the plasma Aβ and AD progression and, also, to confirm the diagnostic potential of the plasma Aβ prior to the plaque formation. We previously found that soluble monomeric Aβ(1–42) in the brain could pass the blood-brain barrier and can be found in the plasma22. To further confirm our hypothesis regarding the impact of plaque formation, we also assessed the permeability of the blood-brain barrier to insoluble fibrillary Aβ(1–42) in 7-week-old ICR mice and studied whether plasma Aβ(1–42) can reflect the condition of brain Aβ(1–42) even after the plaque deposition.
Results
Age-dependent accumulation of Aβ plaques in 3xTg-AD mouse brains
Here, female 3xTg-AD mice aged 5, 7, 9 and 12 months were selected to study changes in the plasma and CSF Aβ levels since ThS-positive Aβ plaques become detectable in the brain as early as 12 months of age (female; 5-month, n = 32; 7-, 9- and 12-month, n = 33)20. To confirm the absence of ThS-positive Aβ plaque in the brains of transgenic mice prior to Aβ measurements, brain samples from each age group were cyrosectioned and fixed with 4% paraformaldehyde for 72 hours, which is different from widely used protocols in other studies21,23. Then, they were stained with ThS for β-sheet-rich Aβ dense-core plaques, 6E10 antibody for both Aβ diffuse and dense-core plaques, DAPI for overall brain visualization and pS199 for hyperphosphorylated tau. We did not observe neither ThS- nor 6E10-positive plaques in the brain of 5-month-old mice (Fig. 1a). 6E10 immunostaining showed diffuse plaques in the hippocampal regions of 7-, 9- and 12-month-old female 3xTg-AD mice (Fig. 1a,b). In addition to the extracellular 6E10-positive plaques, we observed 6E10-stained neural cells from 7-month-old 3xTg-AD mice (Fig. 1a). Of the ThS stained brain slices, only the 12 month-old 3xTg-AD mice showed ThS-positive, dense-core, Aβ plaque depositions in the brain (Fig. 1b). In addition, we observed accumulations of hyperphosphorylated tau tangles in the brain of 12-month-old 3xTg-AD mice.
Age-dependent Aβ(1–42) concentration changes in the CSF of 3xTg-AD mice
Decreased Aβ levels in CSF is a clinical indication of AD progression. To confirm the alterations of CSF Aβ levels in 3xTg-AD mice mimics those in AD patients, the levels of Aβ(1–42) in the CSF of 5-, 7-, 9- and 12-month-old 3xTg-AD mice were measured with sandwich-ELISA utilizing two anti-Aβ antibodies with different epitopes. The CSF was collected using laboratory-produced capillary tubes with tapered tips as previously described24 and then we measured the levels of Aβ(1–42) and Aβ(1–40). The concentration of Aβ(1–42) in the CSF of 3xTg-AD mice showed an age-dependent decrease with a statistical significance before the age of 12 months (female; 5-month, n = 19; 7-month, n = 31; 9-month, n = 31; 12-month, n = 6, P < 0.0001 for 5-month-old vs. 7-month-old and 7-month-old vs. 9-month-old, Fig. 2a)(13 out of 32 brains from 5-month-old and 27 out of 33 brains from 12-month-old groups were excluded from the results due to saturated values during ELISA readings). After the formation of ThS-positive plaques, the CSF Aβ(1–42) level no longer showed a decrease in the trend. On the contrary, we did not observe distinguishable trend in the age-dependent alterations of CSF Aβ(1–40) levels (P = 0.0049 for 5-month-old vs. 9-month-old, P < 0.0001 for 7-month-old vs. 9-month-old, Fig. 2b). Our results agree with previous studies on the age-dependent decline of CSF Aβ(1–42) levels in both AD patients and animal models.
Age-dependent Aβ(1–42) concentration changes in the plasma of 3xTg-AD mice
In order to study if the plasma Aβ level changes in an age-dependent manner in the 3xTg-AD mice, we measured the concentration of plasma Aβ(1–42) in the aforementioned mice used in the CSF Aβ measurements. The blood was first transferred directly from the vena cava to EDTA-containing tubes and the plasma was isolated using centrifugation. Then we measured the levels of Aβ(1–42) and Aβ(1–40). In contrast to the CSF measurements, we found that plasma Aβ(1–42) levels increased from 5- to 9-month-old 3xTg-AD mice (female; 5-month, n = 32; 7-month, n = 32; 9-month, n = 32, Fig. 3a). Notably, the plasma Aβ(1–42) levels decrease in 12-month-old mice, of which brains only developed ThS-positive dense core plaques and hyperphosphoryated tau tangles (12-month, n = 31, Fig. 3a). All the results were statistically significant by one-way ANOVA followed by Bonferroni’s post-hoc comparisons (P = 0.003 for 5-month-old vs. 7-month-old, P < 0.0001 for 7-month-old vs. 9-month-old, P < 0.0001 for 9-month-old vs. 12-month-old). We observed identical trends in the alterations of plasma Aβ(1-40) to those of plasma Aβ(1–42). Between 5-, 7- and 9-month-old groups, 3xTg-AD mice showed significantly increasing levels of Aβ(1–40) in the plasma, whereas the 12-month-old group showed substantial decline (P < 0.0001 for 5-month-old vs. 9-month-old, P < 0.0001 for 7-month-old vs. 9-month-old, Fig. 3b).
To directly compare the age-dependent alterations in Aβ levels, the Aβ(1–42) and Aβ(1–40) concentrations in the plasma and CSF were plotted onto the same graph. The graph shows that the Aβ(1–42) concentrations in the plasma increases with age while the Aβ(1–42) concentrations in the CSF decreases, indicating inverse correlations between the Aβ(1–42) levels in the plasma and CSF (Fig. 4a). However, as there was no trend in CSF Aβ(1–40) levels, it was difficult to conclude any correlations between the Aβ(1–40) levels in the plasma and CSF (Fig. 4b). Collectively, our study concludes that the plasma Aβ(1–42) increases in age-dependent manner in 3xTg-AD mice and has the diagnostic potential for detecting AD until ThS-positive Aβ plaques form in the brain.
Limited blood-brain barrier transport of fibrillary Aβ
Because the blood-brain barrier is known to be permeable to soluble monomeric Aβ(1–42), plasma Aβ(1–42) is a viable candidate for early AD diagnosis. However, if the changes in plasma Aβ(1–42) levels can serve as a biomarker for AD only prior to the plaque formation in the brain, then the plasma should not be able to reflect the condition of brain Aβ(1–42), once the Aβ peptides become insoluble aggregates. Direct fibrillary Aβ injection to tissue enabled us to study the blood-brain barrier’s permeability to insoluble Aβ by measuring the levels of fibrillary Aβ in the plasma. As we observed ThS-negative diffuse plaques did not have an effect on the plasma levels of Aβ in 3xTg-AD mice, it is important to inject ThS-postive Aβ aggregates. Thus, we performed ThS fluorescence assays to confirm formation of ThS-positive aggregates before the intracerebral (IC) injections (Fig. 5a). Then, we injected ThS-positive fibrillary Aβ(1–42) to the cortex of the brains of 7-week-old ICR mice (male, n = 5) by IC injection and assessed the permeability of the blood-brain barrier to fibrillary Aβ. We selected non-transgenic ICR mice for this study, since Aβ in the CSF of transgenic mice can potentially behave as a confounding factor. Additional 5 male ICR mice received IC injection with vehicle as controls. The blood was collected from the vena cava of the mice 30 minutes after the IC injection to allow sufficient time for fibrillary Aβ to pass the blood-brain barrier. After analyzing the plasma with sandwich-ELISA, no statistically significant differences in the plasma soluble Aβ concentrations of fibrillary Aβ-injected mice and vehicle-injected mice were found (Unpaired student’s t-test, P = 0.629, Fig. 5b). Therefore, the brain-blood barrier was not freely permeable to fibrillary Aβ(1–42) and the plasma Aβ(1–42) could not reflect the condition of brain Aβ(1–42) once they become insoluble plaques.
Discussion
In this study, we found that the plasma Aβ(1–42) concentration increases in an age-dependent manner, while the level of CSF Aβ(1–42) decreases, indicating an inverse correlation between the plasma and CSF Aβ(1–42) levels in 3xTg-AD mice before dense-core Aβ plaque depositions appear in their brains. On the contrary, we did not observe such correlation in the alterations of Aβ(1-40) concentration between CSF and plasma. These results suggest that measuring the plasma Aβ(1–42) levels can function as an early diagnostic marker of AD. One previous comparison study involving the APP23 transgenic mice and the TG2576 mice also reported increasing plasma Aβ concentrations in age-dependent manner25 and another comparison study reported that in the amyloid precursor protein transgenic mouse models, Aβ concentrations in CSF decreases when Aβ deposition starts to appear15, further supporting our inverse correlation of Aβ levels. Although other additional studies have investigated Aβ levels in the CNS, they did not directly compare the Aβ levels in the CSF and plasma before and after plaque formation. Our study has further determined the aforementioned issue.
Our results indicate that the plasma Aβ(1–42) concentration possesses a diagnostic potential as a biomarker for early diagnosis of AD when there is no ThS positive Aβ plaque depositions in the brain. This finding is important because it could provide a possible explanation for controversial results from previous studies on the Aβ measurements in blood. Although the plasma Aβ was thought to be related to AD progression and was suggested as a potential target for AD diagnosis, previous controversial results made it difficult to draw a meaningful conclusion. Our study shows that the plasma Aβ(1–42) levels increases with age and, therefore, it can be used as an early marker for AD progression. However, the plasma Aβ may not be an ideal biomarker for AD diagnosis in later developing stages of AD, since amyloid plaque formation makes the plasma Aβ measurements unreliable. The plasma Aβ(1–42) levels may become unable to reflect the AD prognosis after the plaque deposition because insoluble Aβ(1–42) cannot pass the blood-brain barrier as seen in this study. However, one study reported possible endothelial damages to the blood-brain barrier by Aβ(1–42) peptides, which will impair the barrier’s function and increase its permeability26. Therefore, further study is recommended to warrant the diagnostic potential of plasma Aβ(1–42) levels after the plaque deposition in the brain.
It is not clear yet how the formation of amyloid plaques in the brain affects the age-dependent correlations between the plasma and CSF Aβ levels that we observed. Moreover, we only observed such correlations in female transgenic mice, which are known to develop amyloid plaques earlier than males27,28. Thus, the underlying mechanism causing the inverse correlation between the plasma and CSF Aβ(1–42) levels needs to be further studied and investigations with the Dominantly Inherited Alzheimer Network are desired for more clinical data29. Nonetheless, this study opens up more therapeutic strategies for AD patients and could potentially lead to development of more convenient and economic diagnosis of AD in its early stages.
Methods
Materials
Zoletil® (Virbac, France) and Rompun® (Bayer Pharma, Germany) were purchased from SMP animal medicine (Korea). Protease inhibitor cocktail was purchased from Roche Diagnostics (USA). EDTA treated BD vacutainer® was purchased from Becton, Dickinson and Company (USA). Human Aβ42 Ultrasensitive and Human Aβ40 ELISA Kit was purchased from Invitrogen (USA).
Animals
B6;129-Psen1tm1Mpm Tg (APPswe, TauP301L) 1Lfa/Mmjax mice (3xTg-AD) were obtained from Jackson Laboratory (USA) and then bred in a laboratory animal breeding room at the Korea Institute of Science and Technology. The mice were housed in groups of four per cage and maintained at constant temperature with an alternating 12-hour light-dark cycle. Food and water were available ad libitum. 129 mice were assessed in this study; 5-month-old 3xTg-AD (n = 32), 7-month-old 3xTg-AD (n = 33), 9-month-old 3xTg-AD (n = 33) and 12-month-old 3xTg-AD (n = 33). All animal experiments were performed in accordance with the National Institutes of Health guide for the care and use of laboratory animals (NIH Publications No. 8023, revised 1978). The animal studies were approved by the Institutional Animal Care and Use Committee of Korea Institute of Science and Technology (AP-2011L1015).
CSF and plasma collection
CSF collection was performed according to the method described previously24. Mice were anesthetized with a blend of tiletamine.HCl, zolazepam.HCl (80 mg/kg, IP, Zoletil 50®, Virbac, France) and xylazine (20 mg/kg, IP, Rompun®, Bayer Pharma, Germany). The anesthetized mouse was placed horizontally and its cisterna magna was surgically open. The opened meninges were stabbed with laboratory-produced capillary tube that had a tapered tip and achieved CSF. After CSF collection, the mouse was placed supinely and its abdominal cavity was opened. Blood sampling from the vena cava was injected to EDTA tube with protease inhibitor cocktail (Roche Diagnostics, Switzerland, cat# 11836170001) and shaken gently. Plasma was separated from the blood. After centrifugation (3000 rpm, 15 minutes, 4 °C), CSF and plasma samples were stored at −80 °C freezer until use.
ThS staining and immunohistochemistry
Animal were perfused with 0.9% NaCl and the brains were fixed with 4% paraformaldehyde for 3 days and then were immersed in 30% sucrose solution for 3 days23,30. The brain samples were sliced at 35 μm using Cryostat (Microm HM 525, Thermo Scientific, USA) and mounted onto glass slides. Aβ plaques in cryo-sectioned brains were visualized by ThS staining. ThS was suspended in 50% of ethanol at 500 μM and brain sections were stained for 7 minutes. Then, to remove non-specific binding of ThS dye, the sections were rinsed through 100, 95 and 90% of ethanol for 10 seconds each and moved into PBS in succession.
To detect diffused plaque, brain slices were incubated in 0.3% PBS-T for permeabilization. After washing with PBS, brain slices underwent blocking step by 5% BSA for 1 hour and then incubated with primary antibody against Aβ (1:200, 6E10 clone, Covance, USA) overnight at 4 °C. Next day, brain slices were stained with Cy3 conjugated secondary antibody (1:400, Jackson ImmunoResearch, USA). To detect endogenous tau phosphorylation, brain slices were stained with primary antibody against pSer199 (1:200, Abcam, UK). Alexa fluor 633 congugated secondary antibody (1:500, Abcam, UK) was used for fluorocente detection. All slices were stained with Hoechst for counter staining. The images were taken with Leica DM2500 fluorescence microscope and the Cari Zeiss LSM700 confocal microscope.
Aβ(1–42) and Aβ(1–40) analyzed by sandwich-ELISA in mice CSF and plasma
Aβ(1–42) levels in the CSF and plasma were quantified by Aβ42-ultra-sandwich-ELISA kit (Invitrogen, USA, cat# KHB3544) and Aβ(1–40) levels in the CSF and plasma were quantified by Aβ40-Human-ELISA kit (Invitrogen, USA, cat# KHB3482). The procedure was performed according to the manufacturer’s instructions. For the measurement of both Aβ(1–42) and Aβ(1–40) in the plasma samples were diluted 10-fold respectively. To measure Aβ(1–42) and Aβ(1–40) levels in CSF, CSF samples were diluted 2000-fold for Aβ(1–42) and 60-fold for Aβ(1–40), respectively. For all sample dilutions during the analysis, Standard Diluent Buffer was used. Diluted CSF and plasma samples were pipetted into each kit that NH2-terminus of human Aβ specific monoclonal antibody coated wells and co-incubated with each Aβ specific monoclonal antibody. The intensity of this color was directly proportional to the concentration of human Aβ present in the CSF and plasma. All procedures were performed according to the manufacturer’s instructions same as both ELISA kits.
Intracerebral injection of fibrillary Aβ(1–42)
ICR mice were obtained from ORIENT BIO Incorporated (Korea). Fibrillary Aβ(1–42) were prepared from the laboratory-produced synthetic Aβ(1–42) with 10% dimethyl sulfoxide (DMSO) that allow aggregation for 2 weeks at 37 °C and then centrifuged at 15,000 rpm for 5 minutes. After centrifugation, we discarded the supernatant and dissociated the pellet in 10% DMSO as the initial volume. Five 7-week-old ICR mice were IC injected with 5.85 μg fibrillary Aβ(1–42) in 1.3 μL according to previously published procedures31,32. 30 minutes after the IC injection, blood samples from the vena cava were collected and the plasma was separated from the blood. Soluble Aβ(1–42) levels in the plasma were quantified by Aβ42-ultra-sandwich-ELISA kit (Invitrogen, cat# KHB3544). The procedure was performed according to the manufacturer’s instructions as described previously.
Fibrillary Aβ(1–42) detection
Quantitative aggregation of fibrillary Aβ(1–42) was monitored by ThS fluorescence assay. Mixtures of aggregated fibrillary Aβ(1–42) in PBS containing 2 μM ThS (Sigma, USA) were transferred to a black 384-well plate. ThS signal was measured in a Flexstation3 spectrophotometer (Molecular Devices, USA) with an excitation wavelength of 430 nm.
Statistical analysis
Graphs were obtained with GraphPad Prism 5 and statistical analyses were performed with one-way ANOVA followed by Bonferroni’s post-hoc comparisons and Student’s t-test (*P < 0.05, **P < 0.01, ***P < 0.001; other comparisons were not significant). The error bars represent the SEMs. Results from statistical analyses are included in the (supporting information Table S1, S2).
Additional Information
How to cite this article: Cho, S. M. et al. Age-dependent inverse correlations in CSF and plasma amyloid-β(1–42) concentrations prior to amyloid plaque deposition in the brain of 3xTg-AD mice. Sci. Rep. 6, 20185; doi: 10.1038/srep20185 (2016).
References
Graff-Radford, N. R. et al. Association of low plasma Abeta42/Abeta40 ratios with increased imminent risk for mild cognitive impairment and Alzheimer disease. Arch Neurol 64, 354–362, doi: 10.1001/archneur.64.3.354 (2007).
Schupf, N. et al. Peripheral Abeta subspecies as risk biomarkers of Alzheimer’s disease. Proc Natl Acad Sci USA 105, 14052–14057, doi: 10.1073/pnas.0805902105 (2008).
Schupf, N. et al. Change in plasma Ass peptides and onset of dementia in adults with Down syndrome. Neurology 75, 1639–1644, doi: 10.1212/WNL.0b013e3181fb448b (2010).
Seppala, T. T. et al. Plasma Abeta42 and Abeta40 as markers of cognitive change in follow-up: a prospective, longitudinal, population-based cohort study. J Neurol Neurosurg Psychiatry 81, 1123–1127, doi: 10.1136/jnnp.2010.205757 (2010).
Yaffe, K. et al. Association of plasma beta-amyloid level and cognitive reserve with subsequent cognitive decline. JAMA 305, 261–266, doi: 10.1001/jama.2010.1995 (2011).
Koyama, A. et al. Plasma amyloid-beta as a predictor of dementia and cognitive decline: a systematic review and meta-analysis. Arch Neurol 69, 824–831, doi: 10.1001/archneurol.2011.1841 (2012).
Locascio, J. J. et al. Plasma amyloid beta-protein and C-reactive protein in relation to the rate of progression of Alzheimer disease. Arch Neurol 65, 776–785, doi: 10.1001/archneur.65.6.776 (2008).
Sundelof, J. et al. Plasma beta amyloid and the risk of Alzheimer disease and dementia in elderly men: a prospective, population-based cohort study. Arch Neurol 65, 256–263, doi: 10.1001/archneurol.2007.57 (2008).
van Oijen, M. et al. Plasma Abeta(1-40) and Abeta(1-42) and the risk of dementia: a prospective case-cohort study. Lancet Neurol 5, 655–660, doi: 10.1016/S1474-4422(06)70501-4 (2006).
Mehta, P. D. et al. Plasma and cerebrospinal fluid levels of amyloid beta proteins 1-40 and 1-42 in Alzheimer disease. Arch Neurol 57, 100–105, doi: 10.1001/archneur.57.1.100. (2000).
McKhann, G. et al. Clinical diagnosis of Alzheimer’s disease: report of the NINCDS-ADRDA Work Group under the auspices of Department of Health and Human Services Task Force on Alzheimer’s Disease. Neurology 34, 939–944, doi: http://dx.doi.org/10.1212/WNL.34.7.939 (1984).
Jack, C. R., Jr. et al. Serial PIB and MRI in normal, mild cognitive impairment and Alzheimer’s disease: implications for sequence of pathological events in Alzheimer’s disease. Brain 132, 1355–1365, doi: 10.1093/brain/awp062 (2009).
Jack, C. R., Jr. et al. Hypothetical model of dynamic biomarkers of the Alzheimer’s pathological cascade. Lancet Neurol 9, 119–128, doi: 10.1016/S1474-4422(09)70299-6 (2010).
Maia, L. F. et al. Changes in amyloid-beta and Tau in the cerebrospinal fluid of transgenic mice overexpressing amyloid precursor protein. Sci Transl Med 5, 194re192, doi: 10.1126/scitranslmed.3006446 (2013).
Maia, L. F. et al. Increased CSF Abeta during the very early phase of cerebral Abeta deposition in mouse models. EMBO Mol Med 7, 895–903, doi: 10.15252/emmm.201505026 (2015).
Fagan, A. M. et al. Longitudinal change in CSF biomarkers in autosomal-dominant Alzheimer’s disease. Sci Transl Med 6, 226ra230, doi: 10.1126/scitranslmed.3007901 (2014).
Billings, L. M. et al. Learning decreases A beta*56 and tau pathology and ameliorates behavioral decline in 3xTg-AD mice. J Neurosci 27, 751–761, doi: 10.1523/JNEUROSCI.4800-06.2007 (2007).
Billings, L. M. et al. Intraneuronal Abeta causes the onset of early Alzheimer’s disease-related cognitive deficits in transgenic mice. Neuron 45, 675–688, doi: 10.1016/j.neuron.2005.01.040 (2005).
Gimenez-Llort, L. et al. Modeling behavioral and neuronal symptoms of Alzheimer’s disease in mice: a role for intraneuronal amyloid. Neurosci Biobehav Rev 31, 125–147, doi: 10.1016/j.neubiorev.2006.07.007 (2007).
Oddo, S. et al. Triple-transgenic model of Alzheimer’s disease with plaques and tangles: intracellular Abeta and synaptic dysfunction. Neuron 39, 409–421, doi: 10.1016/S0896-6273(03)00434-3 (2003).
Oh, K. J. et al. Staging of Alzheimer’s pathology in triple transgenic mice: a light and electron microscopic analysis. Int J Alzheimers Dis 2010, 1–24, doi: 10.4061/2010/780102 (2010).
Cho, S. M. et al. Correlations of amyloid-beta concentrations between CSF and plasma in acute Alzheimer mouse model. Sci Rep 4, 6777, doi: 10.1038/srep06777 (2014).
Kim, H. Y. et al. EPPS rescues hippocampus-dependent cognitive deficits in APP/PS1 mice by disaggregation of amyloid-beta oligomers and plaques. Nat Commun 6, 8997, doi: 10.1038/ncomms9997 (2015).
Liu, L. & Duff, K. A technique for serial collection of cerebrospinal fluid from the cisterna magna in mouse. J Vis Exp, doi: 10.3791/960 (2008).
Kuo, Y. M. et al. The evolution of A beta peptide burden in the APP23 transgenic mice: implications for A beta deposition in Alzheimer disease. Mol Med 7, 609–618 (2001).
Jancso, G. et al. Beta-amyloid (1-42) peptide impairs blood-brain barrier function after intracarotid infusion in rats. Neurosci Lett 253, 139–141, doi: 10.1016/S0304-3940(98)00622-3 (1998).
Carroll, J. C. et al. Sex differences in beta-amyloid accumulation in 3xTg-AD mice: role of neonatal sex steroid hormone exposure. Brain Res 1366, 233–245, doi: 10.1016/j.brainres.2010.10.009 (2010).
McCullough, L. D. et al. NIH initiative to balance sex of animals in preclinical studies: generative questions to guide policy, implementation and metrics. Biol Sex Differ 5, 15, doi: 10.1186/s13293-014-0015-5 (2014).
Morris, J. C. et al. Developing an international network for Alzheimer research: The Dominantly Inherited Alzheimer Network. Clin Investig (Lond) 2, 975–984, doi: 10.4155/cli.12.93 (2012).
Kim, H. Y. et al. Taurine in drinking water recovers learning and memory in the adult APP/PS1 mouse model of Alzheimer’s disease. Sci Rep 4, 7467, doi: 10.1038/srep07467 (2014).
Eisele, Y. S. et al. Induction of cerebral beta-amyloidosis: intracerebral versus systemic Abeta inoculation. Proc Natl Acad Sci USA 106, 12926–12931, doi: 10.1073/pnas.0903200106 (2009).
Wilcock, D. M. et al. Intracranially administered anti-Abeta antibodies reduce beta-amyloid deposition by mechanisms both independent of and associated with microglial activation. J Neurosci 23, 3745–3751 (2003).
Acknowledgements
This work was supported by KHIDI (HI14C3319) and NST (2N41690, Dementia DTC Project). The authors thank Ms. BoRam Kang (KIST, UST) for IHC staining and imaging advice.
Author information
Authors and Affiliations
Contributions
S.M.C., H.Y.K., T.S.K. and Y.K. designed the experiments. H.V.K., Y.C. and J.W. performed animal preparation. S.L. and S.M.C. prepared brain, C.S.F. and plasma samples. S.M.C., H.Y.K., S.L. and S.H.Y. performed sandwich-ELISA. S.M.C., S.H.Y. and J.K. performed staining and data analysis. S.B. prepared synthetic Aβ. D.K. and Y.K.K. performed I.C. injection. M.J.L., S.M.C., J.Y., S.L., S.H.Y. and Y.K. wrote the manuscript.
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Electronic supplementary material
Rights and permissions
This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
About this article
Cite this article
Cho, S., Lee, S., Yang, SH. et al. Age-dependent inverse correlations in CSF and plasma amyloid-β(1–42) concentrations prior to amyloid plaque deposition in the brain of 3xTg-AD mice. Sci Rep 6, 20185 (2016). https://doi.org/10.1038/srep20185
Received:
Accepted:
Published:
DOI: https://doi.org/10.1038/srep20185
This article is cited by
-
Natural aging and Alzheimer’s disease pathology increase susceptibility to focused ultrasound-induced blood–brain barrier opening
Scientific Reports (2023)
-
Differential effects of chronic immunosuppression on behavioral, epigenetic, and Alzheimer’s disease-associated markers in 3xTg-AD mice
Alzheimer's Research & Therapy (2021)
-
Peripheral adaptive immunity of the triple transgenic mouse model of Alzheimer’s disease
Journal of Neuroinflammation (2019)
-
Beneficial effects of curtailing immune susceptibility in an Alzheimer’s disease model
Journal of Neuroinflammation (2019)
-
Elevated emotional contagion in a mouse model of Alzheimer’s disease is associated with increased synchronization in the insula and amygdala
Scientific Reports (2017)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.