Transcriptome regulation in the brain of a transgenic Alzheimer mouse model treated by lactate
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PROVIDER: PRJNA1157431 | ENA |
REPOSITORIES: ENA
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Project description:To investigate the potential effect of lactate in Alzheimer disease (AD) therapy, we established a transgenic Alzheimer disease mouse model, and analyse the transcriptome regulation induced by lactate compared to vehicle treatment in AD mice. In parallel, the lactate induced transcriptome regulaltion also were analyzed in wild type mice compared to vehicle treatment.
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Project description:We report the application of RNA sequencing technology for high-throughput profiling of brain transcriptomes in ischemic stroke mice. We find that changes in gene expression were detected in vehicle-treated ischemic brains when compared with the sham groups, with 1159 upregulated genes and 529 downregulated genes. While, the lactate preconditioning further enlarged the changes in gene expression of ischemic brains, with 2071 upregulated genes and 1003 downregulated genes. Finally, we show that the ischemic brains are much more immunologically activated by the lactate treatment. This study shows that the contributing roles of lactate in the brain injury after cerebral ischemia are complex and immunological mechanisms may be the major effects that exacerbate the brain injury of mice with cerebral ischemia.
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Project description:L. plantarum is known to possess an L-lactate inducible lactate racemase activity (Goffin et al. 2005. J. Bacteriol. 187:6750). In the present study, microarrays were used in order to identify all genes that are up-regulated by L-lactate, but not by a racemic mixture of D- and L-lactate. A mutant of L. plantarum NCIMB8826 deficient for NAD-dependent L-lactate activity (TF101; Ferain et al. 1994. 176:596), and thus producing no L-lactate, was grown in MRS medium at 28°C until mid-exponential phase (OD600nm 0.75). The culture was then divided into 3 sub-cultures. Optically pure sodium L-lactate (200 mM) was added to the first sub-culture (TF101 + L-lac 200 mM). An equimolar mixture of sodium D- and L-lactate (100 mM each) was added to the second sub-culture (TF101 + L/D-lac 200 mM). The third sub-culture was not treated (TF101; reference sample). The three sub-cultures were further incubated at 28°C for 1h30 (a time known to be sufficient for induction of lactate racemase activity by L-lactate). Cells were harvested by centrifugation. Microarray data were used ot identify genes that are specifically induced by L-lactate (comparison of TF101 with TF101 + L-lac 200 mM), but not by DL-lactate (comparison of TF101 with T101 + L/D-lac 200 mM). There are no biological replicates.
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Project description:Synaptic dysfunction occurs early in Alzheimer´s disease. However, efforts to protect against these detriments have not been possible due to the inadequate understanding of synaptic signalling pathways involved in the neuropathological disease process. The aim of this study was to elucidate the molecular mechanisms behind Alzheimer´s and synaptic plasticity-related signalling using a multi-omics and in situ imaging approach. We used an Alzheimer´s mouse model (APPswe/PSN1dE9, age of 12 month) (APP/PS1) and compared the changes on the proteome including global phosphorylation and N-linked glycosylation pattern, pathway-focused transcriptome and neurological disease-associated miRNAome with age-matched control mice in the neocortex, hippocampus, olfactory bulb and brainstem. Our analysis showed that signalling pathways related to synaptic functions associated to dendritic spine morphology, neurite outgrowth, long-term potentiaiton, CREB signalling and cytoskeletal dynamics were altered in the APP/PS1 transgenic mice, particular in the neocortex and olfactory bulb. This was associated with Aβ plaques and neurofibrillary tangle formation as well as microglial clustering in all brain regions except brainstem. The responses may be epigenetically modulated by the interaction with a number of miRNAs. We suggest that the alterations in synaptic plasticity-related signalling are associated to neurocognitive dysfunctions that resemble the situtation in human AD patients.
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Project description:To investigat the role of AARS1 in HGC27 cells treated with lactate, we transfected HGC27 cells with AARS1 siRNA and then treated the cells with 25mM lactate for 24h
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Project description:CD8+ T cells infiltrate virtually every tissue to find and destroy infected or mutated cells. They often traverse varying oxygen levels and nutrient-deprived microenvironments. High glycolytic activity in tissues can result in extended exposure of cytotoxic T cells to the metabolite lactate. Lactate can be immunosuppressive, at least in part due to its association with tissue acidosis. We show here that the lactate anion is well tolerated by CD8+ T cells in pH neutral conditions. We describe how lactate is taken up by activated CD8+ T cells and is capable of displacing glucose as a carbon source. Activation in the presence of a pH neutral form of lactate significantly alters the CD8+ T cell transcriptome, including the expression of key effector differentiation markers such as granzyme B and interferon-gamma. Our studies reveal the novel metabolic features of lactate utilization by activated CD8+ T cells, and highlight the importance of lactate in shaping the differentiation and activity of cytotoxic T cells. Method: CD8+ T cells were purified from mouse splenocytes and activated for 72h with anti-CD3/CD28 dynabeads (Thermo Fisher, 11456D) and 10 U/ml recombinant human IL-2, in either plain media (RPMI 1640 supplemented with 10% Fetal Bovine Serum, 50 µM 2-mercaptoethanol, and 100 U/ml penicillin-streptomycin), or in the presence of 40 mM sodium lactate. After activation, cells were washed twice with PBS and cell pellets were snap-frozen in RLT Plus lysis buffer (Qiagen, #1053393). Total RNA was extracted with the Qiagen’s RNeasy kit according to the manufacturer's instructions. All samples were quality checked with Agilent Tapestation RNA screen tape. To construct libraries suitable for Illumina sequencing, the Illumina TruSeq Stranded mRNA Sample preparation protocol which includes cDNA synthesis, ligation of adapters, and amplification of indexed libraries was used. The yield and quality of the amplified libraries were analysed using Qubit by Thermo Fisher and the Agilent Tapestation. The indexed cDNA libraries were normalised and combined, and the pools were sequenced on the Nextseq 550 for a 50-cycle v2.5 sequencing run, generating 71 bp single-end reads (and 2*10 bp index reads). Fastq files were generated by demultiplexing with bcl2fastq (v2.20.0.422). STAR 2.7.5b was used to map the fastq files to the mouse reference genome (mm10/GRCm38) and to remove PCR duplicates. Uniquely mapped reads were counted in annotated exons using featureCounts v1.5.1. The gene annotations (Mus_musculus.GRCm38.99.gtf) and reference genome were obtained from Ensembl. The count table from featureCounts was imported into R/Bioconductor and differential gene expression was performed using the EdgeR package and its general linear models pipeline.
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Project description:Experiment Description RNA sequencing was performed on Candida albicans wild type cells (SC5314) grown to exponential phase on YNB Lactate, YNB Glucose, or YNB Glucose plus Lactate, and compared to exponential Candida albicans crz1 cells grown on YNB Glucose or YNB Glucose plus Lactate. Three independent experiments were performed.
2016-10-03 | E-MTAB-4579 | biostudies-arrayexpress