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Molecular mechanisms for Krüppel-like factor 13 actions in hippocampal neurons

ABSTRACT: Molecular mechanisms for Krüppel-like factor 13 actions in hippocampal neurons

PROVIDER: PRJNA624112 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR11518813.fastq.gz	Fastqsanger.gz
	SRR11518814.fastq.gz	Fastqsanger.gz
	SRR11518815.fastq.gz	Fastqsanger.gz
	SRR11518816.fastq.gz	Fastqsanger.gz
	SRR11518817.fastq.gz	Fastqsanger.gz

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Molecular mechanisms for Krüppel-like factor 13 actions in hippocampal neurons [ChSP-seq]

Project description:Molecular mechanisms for Krüppel-like factor 13 actions in hippocampal neurons [ChSP-seq]

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Molecular mechanisms for Krüppel-like factor 13 actions in hippocampal neurons [ChSP-seq]

Project description:Krüppel-like factors (KLFs) play key roles in nervous system development and function. Several KLFs are known to promote, and then maintain neural cell differentiation. Our previous work focused on the actions of KLF9 in mouse hippocampal neurons. Here we investigated genomic targets and functions of KLF9’s paralog KLF13, with the goal of understanding how these two closely related transcription factors influence hippocampal cell function, proliferation, survival and regeneration. We engineered the adult mouse hippocampus-derived cell line HT22 to control Klf13 expression with doxycycline. We also generated HT22 Klf13 knock out cells, and we analyzed primary hippocampal cells from wild type and Klf13-/- mice. RNA sequencing showed that KLF13, like KLF9, acts predominantly as a transcriptional repressor in hippocampal neurons and can regulate other Klf genes. Pathway analysis revealed that genes regulated by KLF13 are involved in cell cycle, cell survival, cytoarchitecture regulation, among others. Chromatin-streptavidin sequencing conducted on chromatin isolated from HT22 cells expressing biotinylated KLF13 identified 9506 genomic targets; 79% were located 1 kilobase upstream of transcription start sites. Transfection-reporter assays confirmed that KLF13 can directly regulate transcriptional activity of its target genes. Comparison of the target genes of KLF9 and KLF13 found that they share some functions that were likely present in their common ancestor, but have also acquired distinct functions during evolution. Flow cytometry showed that KLF13 promotes cell cycle progression, and it protects cells from glutamate-induced excitotoxic damage. Taken together, our findings establish novel roles and molecular mechanisms for KLF13 actions in mammalian hippocampal neurons.

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Molecular mechanisms for Krüppel-like factor 13 actions in hippocampal neurons

Project description:This SuperSeries is composed of the SubSeries listed below.

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Molecular mechanisms for Krüppel-like factor 13 actions in hippocampal neurons [RNA-seq]

Project description:Changes in gene expression in HT22 cells after forced expression of KLF13 for 8 hr were analyzed by RNA sequencing

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Cultured hippocampal neurons glial fibrillary acidic protein (GFAP)

Project description:Despite their widespread use as a research model, a comprehensive, quantitative proteomic profile of the cultured hippocampal neurons has remained unexplored. Here, we provide the first global proteomic characterization of primary murine hippocampal neurons cultured for 14 days under near-physiological glucose conditions (2.5 mM). Using label-free mass spectrometry and the Total Protein Approach (TPA), we quantified over 5,500 proteins, revealing a metabolic signature consistent with oxidative phosphorylation and a neuron-specific proteome largely devoid of astrocytic markers. Unexpectedly, glial fibrillary acidic protein (GFAP) was highly abundant in pure neuronal cultures, and its knockdown impaired neuronal viability and morphology. These findings suggest a compensatory role for GFAP in neurons cultured without astrocytic support. Additionally, our data highlight significant underexpression of key synaptic proteins compared to hippocampal tissue, underscoring the limitations of neuron-only culture systems. This study establishes a new molecular reference for hippocampal neurons and offers insights into neuronal adaptation in simplified environments.

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