Center for Research on Reproduction and Women's Health, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
Reproductive Sciences, Conservation Science Wildlife Health, San Diego Zoo Wildlife Alliance, Escondido, CA, USA.
Sci Rep. 2024 Aug 20;14(1):19321. doi: 10.1038/s41598-024-70235-7.
Much remains unknown about the reproductive physiology of southern white rhinoceros (SWR) and the effect of ovarian stimulation prior to ovum pickup (OPU) have not been fully elucidated. Granulosa cells (GC) provide valuable insight into follicle growth and oocyte maturation status. The goals of this study were to evaluate transcriptomic changes in GC from three stages of follicle development and to identify biomarkers possibly associated with follicular growth and maturation as a result of ovarian stimulation. GC collected from SWRs following OPU were assigned stages based upon follicle size. Total RNA was isolated, and cDNA libraries were prepared and sequenced on a NovaSeq 6000. All bioinformatics analyses were performed utilizing the Galaxy web platform. Reads were aligned to CerSimCot1.0, and the manual curation was performed with EquCab3.0. Overall, 39,455 transcripts (21,612 genes) were identified across follicle stages, and manual curation yielded a 61% increase in gene identification from the original annotation. Granulosa cells from preovulatory follicles expressed the highest number of unique transcripts. The following seven biomarkers were determined based upon cluster analysis and patterns of expression: COL1A1, JMY, FBXW11, NRG1, TMPO, MACIR and COL4A1. These data can be used to potentially evaluate the effects of different ovarian stimulation protocols on follicle dynamics, improve OPU results, and support conservation efforts in this species.
关于南方白犀牛（SWR）的生殖生理学，仍有许多未知之处，而卵巢刺激对卵母细胞采集（OPU）前的影响尚未完全阐明。颗粒细胞（GC）为卵泡生长和卵母细胞成熟状态提供了有价值的见解。本研究的目的是评估来自卵泡发育三个阶段的 GC 的转录组变化，并鉴定可能与卵巢刺激引起的卵泡生长和成熟相关的生物标志物。根据卵泡大小将 OPU 后从 SWR 中收集的 GC 分配到不同阶段。分离总 RNA，并在 NovaSeq 6000 上制备和测序 cDNA 文库。所有生物信息学分析均利用 Galaxy 网络平台进行。将读数与 CerSimCot1.0 对齐，并利用 EquCab3.0 进行手动注释。总体而言，在卵泡阶段鉴定了 39455 个转录本（21612 个基因），而手动注释使原始注释中的基因鉴定增加了 61%。促黄体激素释放激素前卵泡的颗粒细胞表达了最多数量的独特转录本。根据聚类分析和表达模式确定了以下 7 个生物标志物：COL1A1、JMY、FBXW11、NRG1、TMPO、MACIR 和 COL4A1。这些数据可用于潜在地评估不同卵巢刺激方案对卵泡动力学的影响，提高 OPU 结果，并支持该物种的保护工作。
