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Single cell transcriptional profiling of cultured developing neurons upon inflammatory challenge [in vitro]

ABSTRACT: Single cell transcriptional profiling of cultured developing neurons upon inflammatory challenge [in vitro]

PROVIDER: PRJNA541482 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR9023020.fastq.gz	Fastqsanger.gz
	SRR9023030.fastq.gz	Fastqsanger.gz
	SRR9023021.fastq.gz	Fastqsanger.gz
	SRR9023022.fastq.gz	Fastqsanger.gz
	SRR9023023.fastq.gz	Fastqsanger.gz

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Single cell transcriptional profiling of cultured developing neurons upon inflammatory challenge

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Project description:The communication between immune and central nervous system represents an emerging field in neuroscience with many potential consequences both in basic and translational medicine. Among the plethora of mechanisms which might be involved, it is now clear that that immune molecules released upon inflammatory state can modulate different physiological aspects of brain development, resulting in long-lasting pathological consequences which may interfere with normal behavior during adulthood. A common aspect of this immune molecule is to promote deep transcriptional changes responsible for long-term effects. Single-cell RNA sequencing is now becoming a cutting-edge technology for studying the genomic rearrangement occurring at single cell level in different experimental models, thus opening a new frontier in medicine. Here, by using primary cultured neurons as a simplified model for studying neuronal development, we study the long-term consequences of the proinflammatory cytokine interleukine-6 (IL-6) on developing neurons through a single-cell sequencing approach. Based on the immuneprofiling of the analyzed cells, we identified different neuronal and non-neuronal cell clusters whose transcriptional profile changed specifically upon the inflammatory stimulation.

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Single cell transcriptional profiling of cultured developing neurons upon inflammatory challenge

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Cytoplasmic accumulation and aggregation of TDP-43 upon proteasome inhibition in cultured neurons.

Project description:Amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD) are characterized by intraneuronal deposition of the nuclear TAR DNA-binding protein 43 (TDP-43) caused by unknown mechanisms. Here, we studied TDP-43 in primary neurons under different stress conditions and found that only proteasome inhibition by MG-132 or lactacystin could induce significant cytoplasmic accumulation of TDP-43, a histopathological hallmark in disease. This cytoplasmic accumulation was accompanied by phosphorylation, ubiquitination and aggregation of TDP-43, recapitulating major features of disease. Proteasome inhibition produced similar effects in both hippocampal and cortical neurons, as well as in immortalized motor neurons. To determine the contribution of TDP-43 to cell death, we reduced TDP-43 expression using small interfering RNA (siRNA), and found that reduced levels of TDP-43 dose-dependently rendered neurons more vulnerable to MG-132. Taken together, our data suggests a role for the proteasome in subcellular localization of TDP-43, and possibly in disease.

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