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      Chronic Visual Stimulation with LED Light Flickering at 24, 40, or 80 Hz Failed to Reduce Amyloid β Load in the 5XFAD Alzheimer’s Disease Mouse Model

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          Abstract

          A single 1-h session (or 7 d of daily 1-h sessions) of noninvasive visual stimulation with LED light flickering at 40 Hz, but not at 20 or 80 Hz, was reported to increase microglial size and decrease amyloid β (Aβ) load in the 5xFAD mouse model of Alzheimer’s disease. To achieve better therapeutic benefits, we explored the effects of daily 1-h sessions of visual stimulation with continuous light or LED light flickering at 24, 40, or 80 Hz for a period of five weeks in 5xFAD mice. As expected, 33-week-old 5xFAD mice but not control wild-type mice of the same age exhibited an abundance of swollen microglia and Aβ plaques in the visual cortex and hippocampus. Unexpectedly, however, compared with similar session of stimulation with continuous light or a light flickering at 24 or 80 Hz, daily sessions of stimulation with LED light flickering at 40 Hz for five weeks failed to further increase the microglial size and could not noticeably decrease the Aβ load in the visual cortex and hippocampus of the 5xFAD mice. In conclusion, contrary to previous findings based on shorter treatment periods, our data showed that daily noninvasive exposure to a light flickering at 40 Hz for a period of five weeks is not effective in reducing Aβ load in the 5xFAD mouse model of Alzheimer’s disease.

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          Intraneuronal beta-amyloid aggregates, neurodegeneration, and neuron loss in transgenic mice with five familial Alzheimer's disease mutations: potential factors in amyloid plaque formation.

          Mutations in the genes for amyloid precursor protein (APP) and presenilins (PS1, PS2) increase production of beta-amyloid 42 (Abeta42) and cause familial Alzheimer's disease (FAD). Transgenic mice that express FAD mutant APP and PS1 overproduce Abeta42 and exhibit amyloid plaque pathology similar to that found in AD, but most transgenic models develop plaques slowly. To accelerate plaque development and investigate the effects of very high cerebral Abeta42 levels, we generated APP/PS1 double transgenic mice that coexpress five FAD mutations (5XFAD mice) and additively increase Abeta42 production. 5XFAD mice generate Abeta42 almost exclusively and rapidly accumulate massive cerebral Abeta42 levels. Amyloid deposition (and gliosis) begins at 2 months and reaches a very large burden, especially in subiculum and deep cortical layers. Intraneuronal Abeta42 accumulates in 5XFAD brain starting at 1.5 months of age (before plaques form), is aggregated (as determined by thioflavin S staining), and occurs within neuron soma and neurites. Some amyloid deposits originate within morphologically abnormal neuron soma that contain intraneuronal Abeta. Synaptic markers synaptophysin, syntaxin, and postsynaptic density-95 decrease with age in 5XFAD brain, and large pyramidal neurons in cortical layer 5 and subiculum are lost. In addition, levels of the activation subunit of cyclin-dependent kinase 5, p25, are elevated significantly at 9 months in 5XFAD brain, although an upward trend is observed by 3 months of age, before significant neurodegeneration or neuron loss. Finally, 5XFAD mice have impaired memory in the Y-maze. Thus, 5XFAD mice rapidly recapitulate major features of AD amyloid pathology and may be useful models of intraneuronal Abeta42-induced neurodegeneration and amyloid plaque formation.
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            Gamma frequency entrainment attenuates amyloid load and modifies microglia.

            Changes in gamma oscillations (20-50 Hz) have been observed in several neurological disorders. However, the relationship between gamma oscillations and cellular pathologies is unclear. Here we show reduced, behaviourally driven gamma oscillations before the onset of plaque formation or cognitive decline in a mouse model of Alzheimer's disease. Optogenetically driving fast-spiking parvalbumin-positive (FS-PV)-interneurons at gamma (40 Hz), but not other frequencies, reduces levels of amyloid-β (Aβ)1-40 and Aβ 1-42 isoforms. Gene expression profiling revealed induction of genes associated with morphological transformation of microglia, and histological analysis confirmed increased microglia co-localization with Aβ. Subsequently, we designed a non-invasive 40 Hz light-flickering regime that reduced Aβ1-40 and Aβ1-42 levels in the visual cortex of pre-depositing mice and mitigated plaque load in aged, depositing mice. Our findings uncover a previously unappreciated function of gamma rhythms in recruiting both neuronal and glial responses to attenuate Alzheimer's-disease-associated pathology.
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              Multi-sensory Gamma Stimulation Ameliorates Alzheimer’s-Associated Pathology and Improves Cognition

              We previously reported that inducing gamma oscillations with a non-invasive light flicker (Gamma ENtrainment Using Sensory stimulus, or GENUS) impacted pathology in the visual cortex of Alzheimer’s disease mouse models. Here, we designed auditory tone stimulation that drove gamma frequency neural activity in auditory cortex (AC) and hippocampal CA1. Seven days of auditory GENUS improved spatial and recognition memory and reduced amyloid in AC and hippocampus of 5XFAD mice. Changes in activation responses were evident in microglia, astrocytes, and vasculature. Auditory GENUS also reduced phosphorylated tau in the P301S tauopathy model. Furthermore, combined auditory and visual GENUS, but not either alone, entrained gamma oscillations, produced microglial-clustering responses, and decreased amyloid in medial prefrontal cortex. Whole brain analysis using SHIELD revealed widespread reduction of amyloid plaques throughout neocortex after multi-sensory GENUS. Thus, GENUS can be achieved through multiple sensory modalities with wide-ranging effects across multiple brain areas to improve cognitive function.
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                Author and article information

                Journal
                eNeuro
                eNeuro
                eneuro
                eNeuro
                eNeuro
                Society for Neuroscience
                2373-2822
                4 August 2023
                August 2023
                : 10
                : 8
                : ENEURO.0189-23.2023
                Affiliations
                [1 ]Graduate Institute of Biomedical Sciences, China Medical University , Taichung 404333, Taiwan
                [2 ]Neuroscience and Brain Disease Center, China Medical University , Taichung 404333, Taiwan
                [3 ]Drug Development Center, China Medical University , Taichung 404333, Taiwan
                [4 ]Translational Medicine Research Center, China Medical University Hospital , Taichung 404327, Taiwan
                Author notes

                The authors declare no competing financial interests.

                Author contributions: T.W.L. designed research; Y.L.Y. performed research; T.W.L. contributed unpublished reagents/analytic tools; Y.L.Y. and T.W.L. analyzed data; T.W.L. wrote the paper.

                This work was supported by the Healthy Longevity Global Grand Challenge Research Grant AS-HLGC-110-05, the China Medical University Research Grant CMU102-NSC-S1, the Ministry of Science and Technology Research Grant MOST109-2320-B-039-010, the National Health Research Institutes Research Grant NHRI-EX112-10803NI, and the “Drug Development Center, China Medical University” from The Featured Areas Research Center Program within the framework of the Higher Education Sprout Project by the Ministry of Education in Taiwan.

                Correspondence should be addressed to Ted Weita Lai at ted.weita@ 123456me.com .
                Article
                eN-NRS-0189-23
                10.1523/ENEURO.0189-23.2023
                10408781
                37550065
                1b6f611b-ce70-4f77-a4f6-bacd00fbb077
                Copyright © 2023 Yang and Lai

                This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license, which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

                History
                : 1 June 2023
                : 13 July 2023
                : 23 July 2023
                Page count
                Figures: 4, Tables: 1, Equations: 0, References: 11, Pages: 7, Words: 00
                Funding
                Funded by: Healthy Longevity Global Grand Challenge
                Award ID: AS-HLGC-110-05
                Funded by: China Medical University (CMU), doi 10.13039/501100007300;
                Award ID: CMU102-NSC-S1
                Funded by: Ministry of Science and Technology, Taiwan (MOST), doi 10.13039/501100004663;
                Award ID: MOST109-2320-B-039-010
                Funded by: National Health Research Institutes (NHRI), doi 10.13039/501100004737;
                Award ID: NHRI-EX112-10803NI
                Categories
                3
                Research Article: Negative Results
                Disorders of the Nervous System
                Custom metadata
                August 2023

                40-hz flickering light,5xfad mouse model,alzheimer’s disease,amyloid β,γ entrainment,microglia

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