Absence of cumulus cells during in vitro maturation affects lipid metabolism in bovine oocytes
Absence of cumulus cells during in vitro maturation affects lipid metabolism in bovine oocytes
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DOI：
10.1152/ajpendo.00469.2012
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发表时间：
2013-03-01
影响因子：
5.1
通讯作者：
Uzbekova, Svetlana
中科院分区：
文献类型：
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作者：
Auclair, Sylvain;Uzbekov, Rustem;Uzbekova, Svetlana
Auclair S，Uzbekov R，Elis S，Sanchez L，Kireev I，Lardic L，Dalbies-Tran R，Uzbekova S.牛卵母细胞体外成熟过程中缺乏卵丘细胞对脂质代谢的影响。美国生理学杂志内分泌代谢67：E599-E613，2013年。首次发表于2013年1月15日; doi：10.1152/ajpendo.00469.2012.-卵丘细胞（CC）围绕卵母细胞，并通过调节营养摄入进行代谢偶联。体外成熟（IVM）前去除CC降低牛卵母细胞的发育能力，而不影响核减数分裂成熟。目的是探讨CC对卵母细胞胞质成熟的影响与能量代谢的关系。在无血清代谢优化培养基中进行卵丘封闭（CEO）或裸露（DO）卵母细胞的IVM。透射电子显微镜显示不同的分布之间的中期II DO和CEO的膜结合囊泡和脂滴。通过尼罗红染色，证明DO中总脂质水平显著降低。全球转录组学分析显示，差异表达的基因调节能量代谢，转录和翻译之间的CEO和DO。通过Western blot，在卵母细胞和CC中检测到脂肪酸合成酶（FAS）和脂肪酸敏感磷脂酶（HSL）蛋白，表明局部脂肪生成和溶解。DO中FAS蛋白的丰度明显低于CEO中，而CC中的表达高于卵母细胞。相反，HSL蛋白在卵母细胞中比CC中更丰富。此外，只有在体外成熟后的卵母细胞中才检测到活性Ser（563）-磷酸化的HSL，其水平在CEO和DO中相似。总之，在IVM过程中CC的缺乏影响了卵母细胞的脂质代谢，并导致次优的胞质成熟。因此，CC可以通过调节局部脂肪酸合成和脂解来定向消耗营养储存以提供成熟所需的能量，从而影响卵母细胞。
Auclair S, Uzbekov R, Elis S, Sanchez L, Kireev I, Lardic L, Dalbies-Tran R, Uzbekova S. Absence of cumulus cells during in vitro maturation affects lipid metabolism in bovine oocytes. Am J Physiol Endocrinol Metab 67: E599-E613, 2013. First published January 15, 2013; doi:10.1152/ajpendo.00469.2012.-Cumulus cells (CC) surround the oocyte and are coupled metabolically through regulation of nutrient intake. CC removal before in vitro maturation (IVM) decreases bovine oocyte developmental competence without affecting nuclear meiotic maturation. The objective was to investigate the influence of CC on oocyte cytoplasmic maturation in relation to energy metabolism. IVM with either cumulus-enclosed (CEO) or -denuded (DO) oocytes was performed in serum-free metabolically optimized medium. Transmission electron microscopy revealed different distribution of membrane-bound vesicles and lipid droplets between metaphase II DO and CEO. By Nile Red staining, a significant reduction in total lipid level was evidenced in DO. Global transcriptomic analysis revealed differential expression of genes regulating energy metabolism, transcription, and translation between CEO and DO. By Western blot, fatty acid synthase (FAS) and hormone-sensitive phospholipase (HSL) proteins were detected in oocytes and in CC, indicating a local lipogenesis and lypolysis. FAS protein was significantly less abundant in DO that in CEO and more highly expressed in CC than in the oocytes. On the contrary, HSL protein was more abundant in oocytes than in CC. In addition, active Ser(563)-phosphorylated HSL was detected in the oocytes only after IVM, and its level was similar in CEO and DO. In conclusion, absence of CC during IVM affected lipid metabolism in the oocyte and led to suboptimal cytoplasmic maturation. Thus, CC may influence the oocyte by orienting the consumption of nutritive storage via regulation of local fatty acid synthesis and lipolysis to provide energy for maturation.