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Single-cell transcriptome profiling reveals the suppressive role of retinal neurons in microglia activation under diabetes mellitus

ABSTRACT: Single-cell transcriptome profiling reveals the suppressive role of retinal neurons in microglia activation under diabetes mellitus

PROVIDER: PRJNA714553 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR15671383_1.fastq.gz	Fastqsanger.gz
	SRR15671383_2.fastq.gz	Fastqsanger.gz
	SRR15671594_1.fastq.gz	Fastqsanger.gz
	SRR15671594_2.fastq.gz	Fastqsanger.gz

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Single-cell transcriptome profiling reveals the suppressive role of retinal neurons in microglia activation under diabetes mellitus

Project description:Diabetic retinopathy, as one of the common complications of diabetes mellitus, is the leading cause of blindness in working-age population. The disease is characterized by damage to retinal vasculature, which is associated with the activation of retina microglial and chronic neurodegeneration. Previous studies have identified the effects of activated microglial on the retinal neurons, but how the microglial activation is regulated by the others is largely unknown. Here, we performed scRNA-seq on the retina of non-human primates with diabetes mellitus, and identified cell-type-specific molecular changes of the six major cell types. By identifying the ligand-receptor expression patterns among different cells, we established the interactome of the whole retina. The data showed that TNF-α signal mediated the activation of microglial through an autocrine manner. We found Muller and amacrine acted through SPP1 signaling and cone acted through TGFβ signaling, to inhibit the genes related to microglial activation. Our study is the first to profile cell-specific molecular changes and the cell-cell interactome of retina under diabetes mellitus, paving a way to dissect the cellular and molecular mechanisms underlying early-stage diabetic retinopathy.

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Project description:We report the use of RNAseq to identify gene changes in isolated retinal microglia after 4 weeks of STZ-induced diabetes. At this early time point in diabetes induction, single retinal cell suspensions were sorted via FACS using a Cd11b-FITC conjugate. RNA was isolated and preamplified using the SMARTSeq v4 low RNA input kit. After library preparation a 50bp single end read was performed at a depth of 19-34 million reads per sample using the Illumina HiSeq. Constructs were mapped to the rat genome and differential expression calculated between the STZ-treated and control groups.

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Project description:Diabetes mellitus (DM) after transplantation remains a crucial clinical problem in kidney transplantation. To obtain insights into molecular mechanisms underlying the development of post-transplant diabetes mellitus (PTDM) and its early impact on glomerular structures, here we comparatively analyze the proteome of histologically normal appearing glomeruli from patients with PTDM from normoglycemic (NG) transplant recipients, and from recipients with pre-existing type 2 DM (PTDM)

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Project description:In order to evaluate the gene expression profile of retinal microglia cells in different age, we purified CD11b-positive microglia from the retinas of wild type C57BL/6 mice at 3, 12, 18, and 24 months age using cell sorting method with flow cytometry. Age-related genes from isolated retinal microglia were performed using 16 Affymetrix GeneChips of Mouse Exon 1.0ST Arrays. Gene expression level between consecutive age groups (i.e. between 3 and 12 months, 12 and 18 months, and 18 and 24 months) was examined to identify microglia relevant aging genes that demonstrated significant changes. We identified a total 719 genes that showed increasing or decreasing more than 1.5-fold change (p<0.05, one-way ANOVA) for at least one of the three inter age-group comparisons. These identified genes were subjected to a hierarchical cluster analysis to visualize trends in differential expression across individual biological repeats in the 4 age groups. The microglia cells were isolated from wild type C57BL/6 mice with microglia cell specific marker CD11b conjugated with FITC using flowcytometry sorting. The aging time point was designed as 4 groups: 3 moth, 12 month, 18 month and 24 month; each group includes 4 repeats. The total RNA was extracted from isolated retinal microglia cells and reverse transcripted to cDNA after amplification and labeling. The gene expression profile was detected with Affymetrix GeneChip of Mouse Exon 1.0ST Arrays

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