Key Laboratory of Animal Genetics, Breeding and Reproduction of Ministry of Agriculture and Rural Affairs, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China.
Int J Mol Sci. 2022 Nov 28;23(23):14888. doi: 10.3390/ijms232314888.
The oviduct is a dynamic reproductive organ for mammalian reproduction and is required for gamete storage, maturation, fertilization, and early embryonic development, and it directly affects fecundity. However, the molecular regulation of prolificacy occurring in estrous periods remain poorly understood. This study aims to gain a better understanding of the genes involved in regulating goat fecundity in the proteome and transcriptome levels of the oviducts. Twenty female Yunshang black goats (between 2 and 3 years old, weight 52.22 ± 0.43 kg) were divided into high- and low-fecundity groups in the follicular (FH and FL, five individuals per group) and luteal (LH and LL, five individuals per group) phases, respectively. The DIA-based high-resolution mass spectrometry (MS) method was used to quantify proteins in twenty oviducts. A total of 5409 proteins were quantified, and Weighted gene co-expression network analysis (WGCNA) determined that the tan module was highly associated with the high-fecundity trait in the luteal phase, and identified NUP107, ANXA11, COX2, AKP13, and ITF140 as hub proteins. Subsequently, 98 and 167 differentially abundant proteins (DAPs) were identified in the FH vs. FL and LH vs. LL comparison groups, respectively. Parallel reaction monitoring (PRM) was used to validate the results of the proteomics data, and the hub proteins were analyzed with Western blot (WB). In addition, biological adhesion and transporter activity processes were associated with oviductal function, and several proteins that play roles in oviductal communication with gametes or embryos were identified, including CAMSAP3, ITGAM, SYVN1, EMG1, ND5, RING1, CBS, PES1, ELP3, SEC24C, SPP1, and HSPA8. Correlation analysis of proteomics and transcriptomic revealed that the DAPs and differentially expressed genes (DEGs) are commonly involved in the metabolic processes at the follicular phase; they may prepare the oviductal microenvironment for gamete reception; and the MAP kinase activity, estrogen receptor binding, and angiotensin receptor binding terms were enriched in the luteal phase, which may be actively involved in reproductive processes. By generating the proteome data of the oviduct at two critical phases and integrating transcriptome analysis, we uncovered novel aspects of oviductal gene regulation of fecundity and provided a reference for other mammals.
输卵管是哺乳动物生殖的一个动态生殖器官,对于配子储存、成熟、受精和早期胚胎发育至关重要,直接影响繁殖力。然而,发情周期中繁殖力的分子调控仍知之甚少。本研究旨在通过输卵管蛋白质组和转录组水平,更好地了解参与调控山羊繁殖力的基因。将 20 只 2-3 岁(体重 52.22±0.43kg)的云山黑山羊母羊分别分为卵泡期(高、低繁殖力组,每组 5 只)和黄体期(高、低繁殖力组,每组 5 只)。采用基于 DIA 的高分辨率质谱(MS)方法定量 20 个输卵管中的蛋白质。共定量到 5409 个蛋白,加权基因共表达网络分析(WGCNA)确定 tan 模块与黄体期高繁殖力性状高度相关,并鉴定出 NUP107、ANXA11、COX2、AKP13 和 ITF140 为枢纽蛋白。随后,在 FH 与 FL 和 LH 与 LL 比较组中分别鉴定到 98 个和 167 个差异丰度蛋白(DAP)。平行反应监测(PRM)验证蛋白质组数据的结果,并通过 Western blot(WB)分析枢纽蛋白。此外,生物黏附和转运活性过程与输卵管功能相关,鉴定到几个在输卵管与配子或胚胎相互作用中发挥作用的蛋白质,包括 CAMSAP3、ITGAM、SYVN1、EMG1、ND5、RING1、CBS、PES1、ELP3、SEC24C、SPP1 和 HSPA8。蛋白质组学和转录组学的相关性分析表明,DAP 和差异表达基因(DEGs)在卵泡期共同参与代谢过程;它们可能为配子接受准备输卵管微环境;黄体期富集了 MAP 激酶活性、雌激素受体结合和血管紧张素受体结合等术语,可能积极参与生殖过程。通过生成两个关键阶段的输卵管蛋白质组数据并整合转录组分析,揭示了输卵管基因调控繁殖力的新方面,为其他哺乳动物提供了参考。