Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, College of Animal Science and Technology, Sichuan Agricultural University, Chengdu 611130, China.
Genes (Basel). 2020 Aug 26;11(9):1001. doi: 10.3390/genes11091001.
Stearoyl-CoA desaturase (SCD) is known to be an important rate-limiting enzyme in the production of monounsaturated fatty acids (MUFAs). However, the role of this enzyme in goose follicular development is poorly understood. To investigate the metabolic mechanism of SCD during goose follicular development, we observed its expression patterns in vivo and in vitro using quantitative reverse-transcription (qRT)-PCR. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to determine a cellular model of SCD function in granulosa cells (GCs) via overexpression and knockdown. qRT-PCR analysis showed that was abundantly expressed in the GC layer, and was upregulated in preovulatory follicles. Peak expression was found in F1 and prehierarchal follicles with diameters of 4-6 mm and 8-10 mm, respectively. We further found that mRNA expression and corresponding enzyme activity occur in a time-dependent oscillation pattern in vitro, beginning on the first day of GC culture. By LC-MS/MS, we identified numerous changes in metabolite activation and developed an overview of multiple metabolic pathways, 10 of which were associated with lipid metabolism and enriched in both the overexpressed and knockdown groups. Finally, we confirmed cholesterol and pantothenol or pantothenate as potential metabolite biomarkers to study SCD-related lipid metabolism in goose GCs.
硬脂酰辅酶 A 去饱和酶(SCD)是生成单不饱和脂肪酸(MUFAs)的关键限速酶。然而,该酶在鹅卵泡发育中的作用尚未阐明。为了研究 SCD 在鹅卵泡发育过程中的代谢机制,我们采用实时定量反转录 PCR(qRT-PCR)技术在体内和体外观察其表达模式。通过过表达和敲低试验,采用液相色谱-串联质谱(LC-MS/MS)检测了 SCD 在颗粒细胞(GCs)中的功能的细胞模型。qRT-PCR 分析表明,在 GC 层中大量表达,在排卵前卵泡中上调。在直径为 4-6mm 和 8-10mm 的 F1 和前等级卵泡中表达量最高。我们进一步发现,mRNA 表达和相应的酶活性在体外呈时间依赖性波动模式,从 GC 培养的第一天开始。通过 LC-MS/MS,我们鉴定了大量代谢物激活的变化,并开发了多个代谢途径的概述,其中 10 个与脂代谢有关,在过表达和敲低组中均有富集。最后,我们证实胆固醇和泛酸或泛酸盐可作为研究鹅 GCs 中 SCD 相关脂代谢的潜在代谢物生物标志物。