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M6A RNA methylation profiling in hippocampus of POCD mice induced by sevoflurane

ABSTRACT: m6A RNA methylation profiling in hippocampus of POCD mice induced by sevoflurane

PROVIDER: PRJEB45220 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR5956172.fastq.gz	Fastqsanger.gz
	ERR5956173.fastq.gz	Fastqsanger.gz
	ERR5956174.fastq.gz	Fastqsanger.gz
	ERR5956175.fastq.gz	Fastqsanger.gz
	ERR5956176.fastq.gz	Fastqsanger.gz

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m6A RNA methylation profiling in hippocampus of POCD mice induced by sevoflurane

Project description:Elderly patients are apt to cognitive impairment and memory loss after surgical operations. This perioperative cerebral damage named postoperative cognitive dysfunction (POCD) is profoundly affected by anesthesia. N6-methyladenosine (m6A) RNA methylation as a widely-studied epigenetic modification to regulate gene expression, however, is never studied in POCD. Initially, fifty 40-week-old outbred female C57BL/6 mice were conducted Morris water maze test by EthoVision XT working system (Noldus, Netherlands) as manufacturer’s instructions. The escaping latent period of each mouse were recorded. Then Thirty mice were randomly selected and given 2% sevoflurane for 4 h in an automatic anesthetic chamber with size of 24 cm*12 cm*18 cm. After natural resuscitation, POCD and non-POCD mice were picked up based on the dynamics of escaping latent period. The other twenty mice received normal air were treated as negative control. Hippocampus were conducted RIP-seq for investigating m6A RNA methylation.

2021-05-31 | E-MTAB-10510 | biostudies-arrayexpress

RNA sequencing of young mice hippocampus after anesthesia

Project description:We performed RNA sequencing for hippocampus of neonatal mice that were exposed to sevoflurane. We treated a group of neonatal mice with 2.5% sevoflurane for 2 hours on day 6, 7, 8, 9 and treated another group on day 6, 7. On day 36, we collected RNA (both total RNA and polysome-associated RNA) from the hippocampus in the aforementioned two groups and the control group (i.e., without sevoflurane treatment). We prepared the sequencing libraries with the collected RNA and sent them to Illumina sequencing platform.

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m6A RNA methylation promotes XIST-mediated transcriptional repression

Project description:XIST is a long non-coding RNA (lncRNA) that mediates transcriptional silencing of X chromosome genes. Here we show that XIST is highly methylated with at least 78 N6-methyladenosine (m6A) residues, a reversible base modification whose function in lncRNAs is unknown. We show that m6A formation in XIST, as well as cellular mRNAs, is mediated by RBM15 and its paralog RBM15B, which bind the m6A-methylation complex and recruit it to specific sites in RNA. This results in methylation of adenosines in adjacent m6A consensus motifs. Furthermore, knockdown of RBM15 and RBM15B, or knockdown of the m6A methyltransferase METTL3 impairs XIST-mediated gene silencing. A systematic comparison of m6A-binding proteins shows that YTHDC1 preferentially recognizes m6A in XIST and is required for XIST function. Additionally, artificial tethering of YTHDC1 to XIST rescues XIST-mediated silencing upon loss of m6A. These data reveal a pathway of m6A formation and recognition required for XIST-mediated transcriptional repression. Three to four biological HEK293T replicates were used to perform iCLIP of endogenous YTH proteins, RBM15, and RBM15B. Crosslinking induced truncations were identified using CIMS-CITS pipeline.

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Single cell discovery of m6A RNA modifications in the hippocampus

Project description:N6-methyladenosine (m6A) is a prevalent and highly regulated RNA modification essential for RNA metabolism and normal brain function. It is particularly important in the hippocampus, where m6A is implicated in neurogenesis and learning. Although extensively studied, its presence and impact in specific cell types remain poorly understood. We investigated m6A in the hippocampus at the single-cell level, revealing a comprehensive landscape of m6A modifications within individual cells.

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Expression data from sevoflurane anesthesia treated rat

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Project description:By performing m6A-seq analysis on dHL-60 human neutrophils infected with Escherichia coli, we want to identify the m6A methylation pattern in neutrophils after bacteria challenge. m6A-seq analysis revealed that the consensus m6A motifs were most significantly enriched within the m6A peaks with typical m6A peak distribution features, and m6A methylation sites were located primarily in the protein coding sequence region and 3′ untranslated region of mRNA transcripts, in neutrophils with bacterial infection.

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Hakai is required for stabilization of core components of the m6A mRNA methylation machinery

Project description:N6-methyladenosine (m6A) is the most abundant internal modification on mRNA which influences most steps of mRNA metabolism and is involved in several biological functions. The E3 ubiquitin ligase Hakai was previously found in complex with components of the m6A methylation machinery in plants and mammalian cells but its precise function remained to be investigated. Here we show that Hakai is a conserved component of the methyltransferase complex in Drosophila and human cells. In Drosophila, its depletion results in reduced m6A levels and altered m6A-dependent functions including sex determination. We show that its ubiquitination domain is required for dimerisation and interaction with other members of the m6A machinery, while its catalytic activity is dispensable. Finally, we demonstrate that the loss of Hakai destabilizes several subunits of the methyltransferase complex, resulting in impaired m6A deposition. Our work adds new functional and molecular insights into the mechanism of the m6A mRNA writer complex.

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Sevoflurane Exposure Alters Long Non-coding RNA and Messenger RNA Profiles in the Neonatal Mouse Hippocampus

Project description:The focus of current study is to investigate the effect of developing sevoflurane exposure on lncRNA profiles and molecular signaling involvement in neurotoxicity. An Expression Microarray assay (V3.0) and data analysis services were provided by Arraystar Inc. (Rockville, MD) to assess and compare the global abundance of 27,427 lncRNAs and 18,855 mRNAs in the hippocampi of mice exposed to either sevoflurane or normal air controls.

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