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猪卵丘细胞在含有低脂肪酸含量和高脂肪酸含量卵泡液的体外成熟过程中脂质积累的不同动态。

Distinct dynamics of lipid accumulation by porcine cumulus cells during in vitro maturation with follicular fluid of low and high fatty acid contents.

作者信息

Malyszka N, Pawlak P, Cieslak A, Szkudelska K, Lechniak D

机构信息

Poznan University of Life Sciences, Department of Genetics and Animal Breeding, Wolynska 33, 60-637, Poznan, Poland.

Poznan University of Life Sciences, Department of Animal Nutrition, Wolynska 33, 60-637, Poznan, Poland.

出版信息

Theriogenology. 2023 Jan 1;195:93-102. doi: 10.1016/j.theriogenology.2022.10.015. Epub 2022 Oct 21.

Abstract

Follicular fluid (FF) is a complex biological medium providing a fully balanced microenvironment for the oocyte. The standard medium for in vitro maturation of porcine oocytes contains 10% of FF which due to its unknown and inconstant composition is a source of a significant variation. This study aimed to investigate whether follicular fluids of significantly different fatty acid contents (standardized follicular fluid) supplemented to porcine IVM media affects lipid metabolism of porcine oocytes and cumulus cells. Two categories of FF from cyclic gilts containing high (H) and low (L) FA content was added to IVM medium for oocytes from prepubertal gilts. Altogether, 521 cumulus oocyte-complexes (oocytes and corresponding cumulus cells) were analyzed for mRNA expression of 7 genes regulating lipid metabolism and selected traits of the lipid droplets (LD). The applied FFs of different FA levels exerted distinct effects on oocytes and cumulus cells (CCs). During IVM oocytes tended to utilize the lipids as demonstrated by the reduced LD number and lipid fluorescence, whereas cumulus cells accumulated lipids as indicated by the increase in LD number, the occupied area and fluorescence level. Changes in cumulus cells were independent of the FA content in the follicular fluid which means an efficient lipid accumulation during IVM. Final analysis including the effect of FA level on LD traits in oocytes and corresponding CCs revealed two distinct patterns. COCs matured in FF of high FA content were characterized by elevated dynamics of lipid accumulation in CCs and stable lipid content in oocytes. In the case of FF with low FA content, CCs accumulated lipids at a significantly lower rate whereas lipid level in oocytes was reduced. The alterations observed in the LD parameters were not accompanied by changes in oocyte nuclear maturation and in transcript level of any from the 7 analyzed genes. In conclusion, fatty acid content of the follicular fluid supplemented to porcine IVM medium affects lipid metabolism in cumulus cells of the maturing oocyte and application of a standardized FF may help to improve the quality of porcine oocytes matured in vitro.

摘要

卵泡液(FF)是一种复杂的生物介质,为卵母细胞提供完全平衡的微环境。猪卵母细胞体外成熟的标准培养基含有10%的卵泡液,由于其成分未知且不稳定,是显著变异的一个来源。本研究旨在探讨添加到猪体外成熟培养基中的脂肪酸含量显著不同的卵泡液(标准化卵泡液)是否会影响猪卵母细胞和卵丘细胞的脂质代谢。将来自发情周期母猪的两类脂肪酸含量高(H)和低(L)的卵泡液添加到初情期前母猪卵母细胞的体外成熟培养基中。总共对521个卵丘卵母细胞复合体(卵母细胞和相应的卵丘细胞)进行了分析,以检测7个调节脂质代谢的基因的mRNA表达以及脂滴(LD)的选定特征。所应用的不同脂肪酸水平的卵泡液对卵母细胞和卵丘细胞(CCs)产生了不同的影响。在体外成熟过程中,卵母细胞倾向于利用脂质,这表现为脂滴数量和脂质荧光减少,而卵丘细胞则积累脂质,这表现为脂滴数量、占据面积和荧光水平增加。卵丘细胞的变化与卵泡液中的脂肪酸含量无关,这意味着在体外成熟过程中脂质有效积累。包括脂肪酸水平对卵母细胞和相应卵丘细胞中脂滴特征的影响的最终分析揭示了两种不同的模式。在高脂肪酸含量的卵泡液中成熟的卵丘卵母细胞复合体的特征是卵丘细胞中脂质积累的动态增加以及卵母细胞中脂质含量稳定。在脂肪酸含量低的卵泡液的情况下,卵丘细胞积累脂质的速度显著较低,而卵母细胞中的脂质水平降低。在脂滴参数中观察到的变化并未伴随着卵母细胞核成熟以及所分析的7个基因中任何一个的转录水平的变化。总之,添加到猪体外成熟培养基中的卵泡液的脂肪酸含量会影响成熟卵母细胞卵丘细胞中的脂质代谢,应用标准化卵泡液可能有助于提高体外成熟的猪卵母细胞的质量。

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