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Alternative splicing in mechanically stretched podocytes as a model of glomerular hypertension

ABSTRACT: Alternative splicing in mechanically stretched podocytes as a model of glomerular hypertension

PROVIDER: PRJNA1156188 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR30534563_1.fastq.gz	Fastqsanger.gz
	SRR30534563_2.fastq.gz	Fastqsanger.gz
	SRR30534564_1.fastq.gz	Fastqsanger.gz
	SRR30534564_2.fastq.gz	Fastqsanger.gz
	SRR30534565_1.fastq.gz	Fastqsanger.gz

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Comparative protein profiling of stretched (high) and non-stretched podocytes

Project description:Podocyte detachment due to mechanical stress is a common issue in hypertension-induced kidney disease. The present study investigated the proteome alterations in mechanically stretched podocytes, a model for glomerular hypertension in comparison to non-stretched cells. Conditionally immortalized differentiated podocytes were seeded on flexible silicon membranes of a six well plate, which was mounted on a manifold connected to a custom-built stretch apparatus (NIPOKA GmbH, Greifswald, Germany), which induced cyclic pressure variations resulting in upward and downward motion of the silicone membranes. Pressure amplitude was chosen to give a maximum up- and downward deflection of the membrane centre of 8 mm (high stretch).

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Alternative splicing in mechanically stretched podocytes as a model of glomerular hypertension

Project description:Alterations in pre-mRNA splicing play an important role in disease pathophysiology. However, the role of alternative splicing (AS) for podocytes in hypertensive nephropathy (HN) has not been investigated. Here we identifiedAS events that play a role in the development and progression of HN. For this, murine podocytes were exposed to mechanical stretch to model HN, after which proteins and mRNA were measured by proteomics, RNA-Seq, and analyzed by several AS detection tools. We found two Shroom3 isoforms that showed significant changes in expression upon mechanical stretch, which was verified by qRT-PCR and in-situ hybridization. Furthermore, we observed an expression switch of two Myl6 isoforms after mechanical stretch. This switch is accompanied by a change in a C-terminally located amino acid sequence. In summary, mechanical stretch of cultured podocytes is an excellent model to simulate hypertensive nephropathy. In depth RNA-Seq analysis disclosed alternative splicing events and differential gene expression, which together with protein regulation improve the understanding of the pathophysiology of hypertension-induced nephropathy.

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