Hessock Emma A, Edwards J Lannett, Schrick F Neal, Payton Rebecca R, Campagna Shawn R, Pollock Abigayle B, Clark Hannah M, Stokes Allyson E, Klabnik Jessica L, Hill Kennedy S, Roberts Samantha R, Hinson Meredith G, Moorey Sarah E
Department of Animal Science, University of Tennessee, Knoxville, TN, United States.
Department of Chemistry, University of Tennessee, Knoxville, TN, United States.
Front Cell Dev Biol. 2023 May 5;11:1156060. doi: 10.3389/fcell.2023.1156060. eCollection 2023.
Preovulatory follicle response to the luteinizing hormone (LH) surge leads to metabolic, molecular, and functional changes in the oocyte and somatic follicular cells from the onset of estrus to ovulation. Follicular fluid contains metabolites, miRNAs, proteins, and hormones that are byproducts of follicular metabolism and support cellular processes of oocyte, cumulus, and granulosa constituents. Numerous studies have highlighted the importance of follicular fluid composition to support fertility, but critical gaps exist toward understanding dynamic modifications in the follicular fluid metabolome from estrous onset to ovulation. The hypothesis was that abundance of follicular fluid metabolites is dependent on follicle progression post LH surge and variability in follicular fluid metabolome profiles indicate key processes required for preparation of the follicle and oocyte for optimal fertility. The objective was to generate preovulatory follicular fluid metabolome profiles and discern differences in the metabolome of preovulatory follicular fluid samples collected at onset of estrus, 11 h post estrous onset, and 18 h post estrous onset. Estrus was synchronized in non-lactating Jersey cows (n=40) and follicular fluid was collected immediately after the first observed standing mount (hr 0) or at approximately h 11 or 18 after the first standing mount. Ultra-High-Performance Liquid Chromatography-High Resolution Mass Spectrometry was performed on preovulatory follicular fluid samples ( = 9 collected at hr 0, 9 at h 11, and 10 at h 18) and a multiple linear model was performed to determine if time post estrous onset impacted metabolite abundance. Metabolites influenced by time post estrous onset were tested for enrichment in KEGG pathways. Ninety metabolites were identified in follicular fluid samples. Twenty metabolites differed in abundance among timepoints post estrous onset ( ≤ 0.05). Pathways corresponding to amino acid and energy metabolism were enriched with metabolites impacted by time post estrous onset (FDR ≤ 0.10). Results from the current study indicate early response to the LH surge to increase bioavailability of amino acids and metabolites used by the cumulus and granulosa cells for energy production and shuttled into the oocyte to support meiotic maturation. Such metabolites may later be used by the ovulatory follicle for protein production.
排卵前卵泡对促黄体生成素(LH)高峰的反应会导致从发情开始到排卵期间卵母细胞和卵泡体细胞发生代谢、分子和功能变化。卵泡液含有代谢物、微小RNA、蛋白质和激素,它们是卵泡代谢的副产品,并支持卵母细胞、卵丘和颗粒细胞成分的细胞过程。许多研究强调了卵泡液成分对维持生育能力的重要性,但在理解从发情开始到排卵期间卵泡液代谢组的动态变化方面仍存在关键差距。研究假设是卵泡液代谢物的丰度取决于LH高峰后卵泡的进展情况,并且卵泡液代谢组谱的变异性表明了卵泡和卵母细胞为实现最佳生育能力而进行准备所需的关键过程。研究目的是生成排卵前卵泡液代谢组谱,并辨别在发情开始时、发情开始后11小时和发情开始后18小时收集的排卵前卵泡液样本代谢组的差异。对非泌乳泽西奶牛(n = 40)进行发情同步处理,在首次观察到站立爬跨后立即(0小时)或在首次站立爬跨后约11小时或18小时收集卵泡液。对排卵前卵泡液样本(0小时收集9份,11小时收集9份,18小时收集10份)进行超高效液相色谱 - 高分辨率质谱分析,并采用多元线性模型来确定发情开始后的时间是否会影响代谢物丰度。对受发情开始后时间影响的代谢物进行KEGG通路富集分析。在卵泡液样本中鉴定出90种代谢物。20种代谢物在发情开始后的不同时间点丰度存在差异(P≤0.05)。与氨基酸和能量代谢相关的通路富含受发情开始后时间影响的代谢物(FDR≤0.10)。本研究结果表明,对LH高峰的早期反应会增加氨基酸和代谢物的生物利用度,这些物质被卵丘和颗粒细胞用于能量产生,并传递到卵母细胞中以支持减数分裂成熟。这些代谢物随后可能被排卵卵泡用于蛋白质合成。