Zhang Peipei, Zhang Hang, Shahzad Muhammad, Kolachi Hubdar Ali, Li Yupeng, Sheng Hui, Zhang Xiaosheng, Wan Pengcheng, Zhao Xueming
State Key Laboratory of Sheep Genetic Improvement and Healthy Breeding, Institute of Animal Husbandry and Veterinary Sciences, Xinjiang Academy of Agricultural and Reclamation Sciences, Shihezi 832061, China.
Tianjin Key Laboratory of Animal Molecular Breeding and Biotechnology, Tianjin Engineering Research Center of Animal Healthy Farming, Institute of Animal Science and Veterinary, Tianjin Academy of Agricultural Sciences, Tianjin 300381, China.
Int J Mol Sci. 2025 Apr 27;26(9):4151. doi: 10.3390/ijms26094151.
The success of assisted reproductive technology is contingent upon the growth potential of embryos post-vitrification process. When compared to in vivo embryos, it has been found that the high intracellular lipid accumulation inside the in vitro-derived embryos results in poor survival during vitrification. Based on this finding, the present study assessed the impact of incorporating forskolin and linoleic acid (FL) entering in vitro culture (IVC) on the embryos' cryo-survival, lipid content, and viability throughout vitrification. Lipid metabolomics and single-cell RNA sequencing (scRNA-seq) techniques were used to determine the underlying mechanism that the therapies were mimicking. It was observed that out of 726 identified lipids, 26 were expressed differentially between the control and FL groups, with 12 lipids upregulated and 14 lipids downregulated. These lipids were classified as Triacylglycerol (TG), Diacylglycerol (DG), Phosphatidylcholine (PC), and so on. A total of 1079 DEGs were detected between the FL and control groups, consisting of 644 upregulated genes and 435 downregulated genes. These DEGs were significantly enhanced in the arachidonic acid metabolism, lipolysis, fatty acid metabolism, cAMP signaling pathway, and other critical developmental pathways. Based on the observation, it was concluded that forskolin and linoleic acid decreased the droplet content of embryos by modulating lipid metabolism, thus enhancing the vitrified bovine embryos' cryo-survival.
辅助生殖技术的成功取决于胚胎在玻璃化处理后的生长潜力。与体内胚胎相比，研究发现体外培养的胚胎细胞内脂质积累过多，导致玻璃化过程中的存活率较低。基于这一发现，本研究评估了在体外培养（IVC）中加入福斯可林和亚油酸（FL）对胚胎冷冻存活率、脂质含量和玻璃化过程中活力的影响。利用脂质代谢组学和单细胞RNA测序（scRNA-seq）技术来确定这些处理所模拟的潜在机制。结果发现，在726种已鉴定的脂质中，对照组和FL组之间有26种脂质表达存在差异，其中12种脂质上调，14种脂质下调。这些脂质被分类为三酰甘油（TG）、二酰甘油（DG）、磷脂酰胆碱（PC）等。FL组和对照组之间共检测到1079个差异表达基因（DEG），其中包括644个上调基因和435个下调基因。这些DEG在花生四烯酸代谢、脂肪分解、脂肪酸代谢、cAMP信号通路和其他关键发育途径中显著增强。基于这些观察结果，得出结论：福斯可林和亚油酸通过调节脂质代谢降低了胚胎的液滴含量，从而提高了玻璃化牛胚胎的冷冻存活率。
