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MALDI Mass Spectrometry Imaging of lipids and gene expression reveals differences in fatty acid metabolism between follicular compartments in porcine ovaries

机译:脂质和基因表达的MALDI质谱成像显示猪卵巢滤泡区之间的脂肪酸代谢差异

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摘要

In mammals, oocytes develop inside the ovarian follicles; this process is strongly supported by the surrounding follicular environment consisting of cumulus, granulosa and theca cells, and follicular fluid. In the antral follicle, the final stages of oogenesis require large amounts of energy that is produced by follicular cells from substrates including glucose, amino acids and fatty acids (FAs). Since lipid metabolism plays an important role in acquiring oocyte developmental competence, the aim of this study was to investigate site-specificity of lipid metabolism in ovaries by comparing lipid profiles and expression of FA metabolism-related genes in different ovarian compartments. Using MALDI Mass Spectrometry Imaging, images of porcine ovary sections were reconstructed from lipid ion signals for the first time. Cluster analysis of ion spectra revealed differences in spatial distribution of lipid species among ovarian compartments, notably between the follicles and interstitial tissue. Inside the follicles analysis differentiated follicular fluid, granulosa, theca and the oocyte-cumulus complex. Moreover, by transcript quantification using real time PCR, we showed that expression of five key genes in FA metabolism significantly varied between somatic follicular cells (theca, granulosa and cumulus) and the oocyte. In conclusion, lipid metabolism differs between ovarian and follicular compartments.
机译:在哺乳动物中,卵母细胞在卵泡内部发育。该过程得到周围卵泡环境的强烈支持,卵泡环境由积云,颗粒细胞和卵泡膜细胞以及卵泡液组成。在窦囊中,卵子形成的最后阶段需要大量的能量,这些能量是由卵泡细胞从包括葡萄糖,氨基酸和脂肪酸(FAs)的基质中产生的。由于脂质代谢在获取卵母细胞发育能力中起着重要作用,因此本研究的目的是通过比较脂质分布图和FA代谢相关基因在不同卵巢区室中的表达来研究卵巢中脂质代谢的位点特异性。使用MALDI质谱成像技术,首次从脂质离子信号重建了猪卵巢切片的图像。离子光谱的聚类分析揭示了卵巢各部分之间,尤其是卵泡和间质组织之间脂质种类的空间分布差异。在卵泡内部分析可区分卵泡液,颗粒,卵泡膜和卵母细胞-卵丘复合体。此外,通过实时PCR的转录定量,我们显示FA代谢中的五个关键基因的表达在卵泡细胞(卵泡膜,颗粒和卵丘)和卵母细胞之间显着不同。总之,卵巢和滤泡区的脂质代谢不同。

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