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Cross-Platform single cell analysis of kidney development shows stromal cells express Gdnf

ABSTRACT: Cross-Platform single cell analysis of kidney development shows stromal cells express Gdnf

PROVIDER: PRJNA412523 | ENA |

REPOSITORIES: ENA

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			Action	DRS
		Other
	SRR6114440_1.fastq.gz	Fastqsanger.gz
	SRR6114440_2.fastq.gz	Fastqsanger.gz
	SRR6114450.fastq.gz	Fastqsanger.gz
	SRR6114460.fastq.gz	Fastqsanger.gz

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Cross-platform single cell analysis of kidney development shows stromal cells express Gdnf.

Project description:The developing kidney provides a useful model for study of the principles of organogenesis. In this report we use three independent platforms, Drop-Seq, Chromium 10x Genomics and Fluidigm C1, to carry out single cell RNA-Seq (scRNA-Seq) analysis of the E14.5 mouse kidney. Using the software AltAnalyze, in conjunction with the unsupervised approach ICGS, we were unable to identify and confirm the presence of 16 distinct cell populations during this stage of active nephrogenesis. Using a novel integrative supervised computational strategy, we were able to successfully harmonize and compare the cell profiles across all three technological platforms. Analysis of possible cross compartment receptor/ligand interactions identified the nephrogenic zone stroma as a source of GDNF. This was unexpected because the cap mesenchyme nephron progenitors had been thought to be the sole source of GDNF, which is a key driver of branching morphogenesis of the collecting duct system. The expression of Gdnf by stromal cells was validated in several ways, including Gdnf in situ hybridization combined with immunohistochemistry for SIX2, and marker of nephron progenitors, and MEIS1, a marker of stromal cells. Finally, the single cell gene expression profiles generated in this study confirmed and extended previous work showing the presence of multilineage priming during kidney development. Nephron progenitors showed stochastic expression of genes associated with multiple potential differentiation lineages.

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Cross-Platform single cell analysis of kidney development shows stromal cells express Gdnf

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Project description:Expression of GDNF-regulated genes was studied in cultures of self-renewing rat spermatogonial stem cells established from 8-10 day old rat pups maintained in a defined serum free medium. GDNF is the primary regulator of spermatogonial stem cell self renewal in the rat. GDNF regulated genes were identified using microarray profiling rat spermatogonial stem cells in the presence and absence of GDNF.

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