| Experiment Id | GSE303512 | Name | Single-cell RNA Sequencing of Wildtype and/or Dazl Mutant Mouse Ovarian Cells Across Embryonic Developmental Stages (E10.5 (Wild type), E11.5 (Wild type and Dazl-/-), E12.5 (Dazl-/-), E15.5 (Wild type) |
| Experiment Type | RNA-Seq | Study Type | Baseline and WT vs. Mutant |
| Source | GEO | Curation Date | 2026-05-27 |
| description | We have identified a Golgi- and microtubule-rich vesicular structure, termed Visham, which resembles the Drosophila fusome and accumulates asymmetrically during germ cell cyst divisions starting at E11.5. Genes associated with Visham become enriched in presumptive oocytes during cyst divisions and early meiosis, suggesting its role as a potential oocyte rejuvenation center. To investigate this further, we performed single-cell RNA sequencing (scRNA-seq) of mouse fetal germ cells at E10.5, E11.5, and E15.5, including developmental stages not previously covered in earlier datasets (GSE136441) and thus generated a composit dataset of E10.5-P5 germ cells. Additionally, we analyzed Dazl homozygous and heterozygous mutant ovarian germ cells at E11.5 and E12.5 to study the impact of Dazl loss on cyst formation and Visham-associated gene expression dynamics. This dataset provides a comprehensive resource to explore transcriptional changes across key stages of fetal ovarian development and in the context of Dazl function. Embryonic ovaries were dissected, and fetal sex was identified using Quinacrine and DAPI staining of tail samples to exclude males in E10.5/E11.5 gonad samples. Female gonads were pooled and dissociated into single cells using 0.25% Trypsin at 37C for 57 minutes with gentle trituration. Trypsin was neutralized with 10% FBS, and cells were filtered through a 30m strainer. After centrifugation and resuspension in 0.04% BSA, cell viability was assessed with trypan blue, and high-quality samples (>90% viability) were loaded (~10,000 cells) onto the 10X (Genomics Chromium platform (v3/v4 chemistry) per manufacturers instructions. For mutant samples, Dazl+/- males and females were mated, and embryos collected at E11.5 and E12.5. Tail DNA was extracted using the KAPA Express Extract Kit and genotyped using standard JAX protocols (strain 035880). Identified homozygous and heterozygous fetal gonads were dissociated as above and subjected to scRNA-seq on the same 10X chromium platform. |
