ABSTRACT: RNA sequencing of ID4-GFP Bright and ID4-GFP Dim spermatogonia from postnatal day 8 transgenic Id4-Gfp (LT11) mouse pups and ID4-GFP+ spermatogonia from ID4 overexpression mouse model and control
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.
Project description:To reveal distinct transcriptomes associated with spermatogonial stem cell renewal vs. initiation of differentiation, single-cell transcriptomes from P6 ID4-EGFP+ spermatogonia (sorted for brightest or dimmest) or unselected testis cells were used for Drop-Seq 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 (a commercial Drop-Seq variant) to perform single-cell RNA-seq
Project description:To reveal distinct transcriptome changes among ID4-EGFP-bright adult mouse spermatogonia associated with mTORC1 activity, single-cell transcriptomes were generated from GFP-bright/CD9-bright spermatogonia from adult mice in three groups: control (untreated), 2 days of Rapamycin treatment (Rapamycin) and 2 days Rapamycin plus 1 day washout (Rapamycin_Release). Based on transplantation studies performed previously, ID4-EGFPbright cells are highly enriched for SSCs. We used the 10x Genomics Chromium to perform single-cell RNA-seq.
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
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
Project description:Detailed FACS analyses of enzymatically disaggregated embryonic hearts from Wt1GFPKI mice revealed two distinct types of Wt1GFP-positive cells based on their GFP expression levels: a Wt1GFP bright population (H) and a Wt1GFP dim (M) population. Quantitative RT-PCR analyses for epicardial genes demonstrated an enrichment in epicardial genes in the Wt1GFP bright population (H)
Project description:To reveal distinct transcriptomes associated with spermatogonial stem cell renewal vs. initiation of differentiation, single-cell transcriptomes from Adult Human spermatogonia were subdivided into subpopulations based on the levels of ID4 mRNA (determined in this experiment). This correlates with distinct fates of corresponding mouse spermatogonia when assayed by transplantation, with ID4-EGFPbright cells highly enriched for SSCs, and ID4-EGFPdim cells enriched for progenitors. We used the Fluidigm C1 instrument to capture individual spermatogonia for SMART-Seq2 single-cell RNA-seq.
Project description:Detailed FACS analyses of enzymatically disaggregated embryonic hearts from Wt1GFPKI mice revealed two distinct types of Wt1-GFP-positive cells based on their GFP expression levels: a Wt1-GFP bright population (H) and a Wt1-GFP dim (M) population. Interestingly, when viewed under the microscope, cultured Wt1GFP+ (M) cells were shown to exhibit synchronous contractions that suggest a cardiomyocyte identity. These observations also imply that a population of low Wt1-expressing cardiomyocytes contributes to the formation of the myocardium.
