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ID4 levels dictate the stem cell state in mouse spermatogonia

ABSTRACT: ID4 levels dictate the stem cell state in mouse spermatogonia

PROVIDER: PRJNA362611 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR5189920_1.fastq.gz	Fastqsanger.gz
	SRR5189920_2.fastq.gz	Fastqsanger.gz
	SRR5189930_1.fastq.gz	Fastqsanger.gz
	SRR5189930_2.fastq.gz	Fastqsanger.gz
	SRR5189921_1.fastq.gz	Fastqsanger.gz

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RNA sequencing of ID4-GFP Bright and ID4-GFP Dim spermatogonia from postnatal day 8 transgenic Id4-Gfp (LT11) mouse pups

Project description:Spermatogonia expressing the highest levels of ID4 (ID4-GFP Bright) represent a population highly enriched for spermatogonial stem cells (SSC) while those expressing lower levels (ID4-GFP Dim) are the putative immediate progenitors. Comparing the transcriptome of these populations can provide insight into the SSC to progenitor transition.

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Project description:Mice that constitutively overexpress ID4 in germ cells have impaired spermatogenic lineage development. The transcriptome of ID4-GFP+ spermatogonia from testes of ID4 overexpression animals was compared to the ID4-GFP+ population from controls

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ID4 levels dictate the stem cell state in mouse spermatogonia.

Project description:Spermatogenesis is a classic model of cycling cell lineages that depend on a balance between stem cell self-renewal for continuity and the formation of progenitors as the initial step in the production of differentiated cells. The mechanisms that guide the continuum of spermatogonial stem cell (SSC) to progenitor spermatogonial transition and precise identifiers of subtypes in the process are undefined. Here we used an Id4-eGfp reporter mouse to discover that EGFP intensity is predictive of the subsets, with the ID4-EGFPBright population being mostly, if not purely, SSCs, whereas the ID4-EGFPDim population is in transition to the progenitor state. These subsets are also distinguishable by transcriptome signatures. Moreover, using a conditional overexpression mouse model, we found that transition from the stem cell to the immediate progenitor state requires downregulation of Id4 coincident with a major change in the transcriptome. Collectively, our results demonstrate that the level of ID4 is predictive of stem cell or progenitor capacity in spermatogonia and dictates the interface of transition between the different functional states.

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Project description:To reveal distinct transcriptomes associated with various spermatogenic cells, including spermatogonial stem cells and all of their subsequent progeny, single-cell transcriptomes from Adult ID4-EGFP+ spermatogonia (sorted for brightest or dimmest), StaPut-enriched spermatocytes and spermatids, or unselected steady-state spermatogenic cells were used for 10x Genomics analysis. The GFP-bright and dim phenotypes exhibit distinct fates when assayed by transplantation, with ID4-EGFPbright cells highly enriched for SSCs, and ID4-EGFPdim cells enriched for progenitors. We used the 10x Genomics Chromium (Drop-Seq) to perform single-cell RNA-seq

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Project description:P6 ID4-EGFP+ undifferentiated spermatogonia, including those stained robustly (high) or weakly (low) for TSPAN8 were isolated by FACS.

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ID4 levels dictate the stem cell state in mouse spermatogonia

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Project description:P6 ID4-EGFP+ undifferentiated spermatogonia stained robustly (high) or weakly (low) for TSPAN8 were isolated by FACS.

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