Journal of Integrative Agriculture

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scRNA-seq揭示山羊排卵过程中卵泡微环境的分子特征

  

  • 修回日期:2024-09-26

scRNA-seq revealing the molecular atlas of the goat follicular microenvironment over the time course of ovulation

Conghui Guoa, b, c, †, Guangbin Liua, b, c†, Jie Liua, b, c, Kaihao Chena, b, c, Ming Denga, b, c, Baoli Suna, b, c, Yongqing Guoa, b, c, Dewu Liua, b, c*, Yaokun Lia, b, c*   

  1. a College of Animal Science, South China Agricultural University, Guangzhou 510642, China;

    b National Local Joint Engineering Research Center of Livestock and Poultry, South China Agricultural University, Guangzhou 510642, China;

    c Lingnan Guangdong Laboratory of Modern Agriculture, Guangzhou, China.

  • Revised:2024-09-26
  • About author:Yaokun Li, ykli@scau.edu.cn; Dewu Liu, dwliu@scau.edu.cn. † These authors contributed equally to this work.
  • Supported by:

    This research was supported by the Guangdong Seed Industry Revitalization Project (2022-XPY-00-010), the Guangdong-Guangxi Cooperative Technology Special Envoys Team Project (E230257), and the Modern Agricultural Industrial Technology System of Guangdong Province (2023KJ127).

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摘要：

哺乳动物的排卵是一个高度复杂且精密调控的生物过程，涉及多种细胞类型、信号分子和细胞因子的协同作用。卵巢卵泡液为排卵提供了适宜的微环境，排卵的顺利进行与雌性动物的生育能力以及农场的经济效益紧密相关。因此，深入了解排卵过程及其分子机制具有重要意义。本研究旨在通过绘制山羊卵泡液的单细胞图谱，探讨排卵的时间动态。我们对hCG刺激后0 h、6 h、12 h、18 h、24 h的山羊卵泡液进行了单细胞转录组测序，鉴定出其中四种细胞类型：颗粒细胞、髓系细胞、T细胞和B细胞，并进一步划分出3个颗粒细胞亚群（生长卵泡颗粒细胞；排卵卵泡颗粒细胞；闭锁卵泡颗粒细胞）和6个髓系细胞亚群（促炎巨噬细胞a；促炎巨噬细胞b；抑炎巨噬细胞；新增巨噬细胞；单核细胞；中性粒细胞）。此外，我们还解析了排卵过程中颗粒细胞的基因表达和功能变化。尤其值得注意的是，我们发现LH峰后的6小时内，大量免疫细胞，特别是巨噬细胞，迅速浸入卵泡液。同时，我们还揭示了颗粒细胞与巨噬细胞之间密切的相互作用，提示颗粒细胞可能在免疫细胞浸润的调节中发挥作用，巨噬细胞可能促进卵母细胞的成熟。总体而言，我们的研究构建了山羊卵泡排卵的单细胞转录组图谱，揭示了排卵过程中细胞类型及基因表达的变化、细胞互作关系，为深入探索排卵机制及排卵障碍的治疗提供了重要基础，为山羊繁殖力的提升、经济效益的扩大提供新思路。

Abstract:

Ovulation is paramount for female animal fertility, necessitating a thorough understanding of its process and molecular underpinnings. This study aimed to delineate the goat ovary temporal dynamics of ovulation. Utilizing single-cell sequencing, we analyzed follicular fluid samples obtained at 0, 6, 12, 18, and 24 h post-hCG administration, identifying 4 cell types and 6 myeloid cell subtypes. We elucidated gene expression and functional changes in granulosa cells over the time course of ovulation. Notably, our study detected and confirmed immune cell infiltration at 6 h post-LH peak. Additionally, we observed significant cellular interactions between granulosa cells and macrophages, with granulosa cells expressing IL1RAP, COL4A1, COL4A2 implicated in immune cell regulation, while macrophages expressed EREG to facilitate oocyte maturation. Collectively, our investigation has established a comprehensive single-cell transcriptome atlas of the ovulating goat ovary for advancing exploration into ovulation mechanisms and developing therapies for ovulatory disorders.

Key words: hCG , ovulation ,  granulosa cell ,  macrophages