Next generation RNA sequencing of Aβ-treated primary cultured mouse astrocytes to study the effect of ODC1 inhibition on AD-like astrocytes
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ABSTRACT:
PROVIDER: PRJNA771360 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR16339857_1.fastq.gz | Fastqsanger.gz | |||
| SRR16339857_2.fastq.gz | Fastqsanger.gz | |||
| SRR16339858_1.fastq.gz | Fastqsanger.gz | |||
| SRR16339858_2.fastq.gz | Fastqsanger.gz | |||
| SRR16339859_1.fastq.gz | Fastqsanger.gz |
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Project description:Reactive astrogliosis is a well-known and recognised marker of Alzheimer's Disease (AD). The aim of this study is to functionally visualise reactive astrocyte-mediated neuronal hypometabolism in the brains that is associated with AD and neuroinflammation. To investigate alterations of acetate and glucose metabolism and the effect thereof in AD-like astrocytes, primary astrocyte cultures were treated with acetate in the presence or absence of amyloid beta (Aβ) oligomers. On comparison, we found that acetate treatment upregulated the expression of acetate transporter MCT1 (Slc16a1) as well as that of enzymes involved in the urea cycle and subsequent GABA synthesis. We could, therefore, conclude that MCT1-mediated uptake of acetate into astrocytes triggered GABA synthesis by upregulating enzymes involved in putrescine production and degradation.
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Project description:The major genetic risk factor for Alzheimer’s disease (AD), APOE4, accelerates beta-amyloid (Aβ) plaque formation, but whether this is caused by APOE expressed in microglia or astrocytes is debated. We express here the human APOE isoforms in astrocytes in an Apoe-deficient AD mouse model. This is not only sufficient to restore the amyloid plaque pathology but also induces the characteristic transcriptional pathological responses in Apoe-deficient microglia surrounding the plaques. We find that both APOE4 and the protective APOE2 from astrocytes increase fibrillar plaque deposition, but differentially affect soluble Aβ aggregates. Microglia and astrocytes show specific alterations in function of APOE genotype expressed in astrocytes. Our experiments indicate a central role of the astrocytes in APOE mediated amyloid plaque pathology and in the induction of associated microglia responses.
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Project description:Astrocytes, one of the most resilient cells in the brain, transform into reactive astrocytes in response to toxic proteins such as amyloid beta (Aβ) in Alzheimer’s disease (AD). However, reactive astrocyte-mediated non-cell autonomous neuropathological mechanism is not fully understood yet. We aimed our study to find out whether Aβ-induced proteotoxic stress affects the expression of autophagy genes and the modulation of autophagic flux in astrocytes, and if yes, how Aβ-induced autophagy-associated genes are involved Aβ clearance in astrocytes of animal model of AD. Whole RNA sequencing (RNA-seq) was performed to detect gene expression patterns in Aβ-treated human astrocytes in a time-dependent manner. To verify the role of astrocytic autophagy in an AD mouse model, we developed AAVs expressing shRNAs for MAP1LC3B/LC3B (LC3B) and Sequestosome1 (SQSTM1) based on AAV-R-CREon vector, which is a Cre recombinase-dependent gene-silencing system. Also, the effect of astrocyte-specific overexpression of LC3B on the neuropathology in AD (APP/PS1) mice was determined. Neuropathological alterations of AD mice with astrocytic autophagy dysfunction were observed by confocal microscopy and transmission electron microscope (TEM). Behavioral changes of mice were examined through novel object recognition test (NOR) and novel object place recognition test (NOPR). Here, we show that astrocytes, unlike neurons, undergo plastic changes in autophagic processes to remove Aβ. Aβ transiently induces expression of LC3B gene and turns on a prolonged transcription of SQSTM1 gene. The Aβ-induced astrocytic autophagy accelerates urea cycle and putrescine degradation pathway. Pharmacological inhibition of autophagy exacerbates mitochondrial dysfunction and oxidative stress in astrocytes. Astrocyte-specific knockdown of LC3B and SQSTM1 significantly increases Aβ plaque formation and hypertrophic reactive astrocytes in APP/PS1 mice, along with a significant reduction of neuronal marker and cognitive function. In contrast, astrocyte-specific overexpression of LC3B reduced Ab aggregates in the brain of APP/PS1 mice An increase of LC3B and SQSTM1 protein is found in astrocytes of the hippocampus in AD patients. Taken together, our data indicates that Aβ-induced astrocytic autophagic plasticity is an important cellular event to modulate Aβ clearance and maintain cognitive function in AD mice.
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Project description:Next-generation RNA sequencing of Acetate treated primary cultured mouse astrocytes in the presence or absence of Aβ
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Project description:To reveal the mechanisms that Ogt controls inflammatory response of astrocytes, we first performed RNA-seq with Ctrl and Ogt cKO astrocytes isolated from the brains of adult mice. Given the inflammation in the brain of Alzheimer’s disease, we next performed RNA-seq with astrocytes isolated from the brains of adult Ctrl and 5X APP Alzheimer’s mice model (AD), and Aβ treated astrocytes, respectively. Taken together, these findings suggest that differentially expressed genes shared between cKO astrocytes, AD astrocytes and Aβ-treated astrocytes are related with inflammation.
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