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.
