College of Animal Sciences and Technology, Qingdao Agricultural University, Qingdao, China.
College of Life Sciences, Qingdao Agricultural University, Qingdao, China.
Reprod Domest Anim. 2021 May;56(5):801-811. doi: 10.1111/rda.13920. Epub 2021 Mar 19.
The new technology of high-throughput single-cell RNA sequencing (10 × scRNA-seq) was developed recently with many advantages. However, it was not commonly used in farm animal research. There are few reports for the gene expression of goat ovarian follicle granulosa cells (GCs) during different developmental stages. In the current investigation, the gene expression of follicle GCs at different stages from two populations of Ji'ning grey goats: high litter size (HL; ≥3/L; 2 L) and low litter size (LL; ≤2 /L; 2 L) were analysed by scRNA-seq. Many GC marker genes were identified, and the pseudo-time showed that GCs developed during the time course which reflected the follicular development and differentiation trajectory. Moreover, the gene expression difference between the two populations HL versus LL was very clear at different developmental stages. Many marker genes differentially expressed at different developmental stages. ASIP and ASPN were found to be highly expressed in the early stage of GCs, INHA, INHBA, MFGE8 and HSD17B1 were identified to be highly expressed in the growing stage of GCs, while IGFBP2, IGFBP5 and CYP11A1 were found to be highly expressed in late stage. These marker genes could be used as reference genes of goat follicle GC development. This investigation for the first time discovered the gene expression patterns in goat follicle GCs in high- or low-fertility populations (based on litter size) by scRNA-seq which may be useful for uncovering the oocyte development potential.
最近开发了高通量单细胞 RNA 测序(10×scRNA-seq)新技术,具有许多优势。然而,它在农场动物研究中并未得到广泛应用。关于不同发育阶段山羊卵巢卵泡颗粒细胞(GCs)的基因表达,仅有少量报道。在本研究中,通过 scRNA-seq 分析了来自两个济宁灰山羊群体(高产羔数(HL;≥3/L;2 L)和低产羔数(LL;≤2/L;2 L))不同阶段卵泡 GCs 的基因表达。鉴定出许多 GC 标记基因,伪时间显示 GCs 在反映卵泡发育和分化轨迹的时间过程中发育。此外,在不同发育阶段,两个群体 HL 与 LL 之间的基因表达差异非常明显。许多标记基因在不同发育阶段的表达存在差异。发现 ASIP 和 ASPN 在 GCs 的早期高度表达,INHA、INHBA、MFGE8 和 HSD17B1 在 GCs 的生长阶段高度表达,而 IGFBP2、IGFBP5 和 CYP11A1 在晚期高度表达。这些标记基因可作为山羊卵泡 GC 发育的参考基因。本研究首次通过 scRNA-seq 发现了高产或低产(基于产羔数)群体中山羊卵泡 GCs 的基因表达模式,这可能有助于揭示卵母细胞发育潜力。