Abstract
ObjectiveTo evaluate human oocyte ability to undergo fertilization and subsequent preimplantation embryonic development in relation to a wide panel of follicular fluid (FF) arachidonic acid derivatives (AAD) and linoleic acid derivatives (LAD) of prospectively selected patients undergoing intracytoplasmic sperm injection (ICSI).MethodologyStudy was designed as a two center (a university clinic and a private clinic) prospective study. 54 women of 181 consecutive couples undergoing ICSI were prospectively found to be eligible for analysis. 'One follicle – one retrieved oocyte – one resulting embryo' approach was used. Each individual follicle was aspirated independently and matched to an oocyte growing in this particular follicular milieu. FF samples were assessed for AAD and LAD by high-performance liquid chromatography; additionally, activity of secretory phospholipase A (sPLA2) was determined by enzyme-linked immunosorbent assay.Principal FindingsIncreased activity of sPLA2 and significantly higher AAD and LAD levels were found in FF of oocytes that did not show two pronuclei or underwent degeneration after ICSI in comparison to oocytes with the appearance of two pronuclei. Receiver operating characteristics curve analysis identified acids with the highest sensitivity and specificity: 5oxo-hydroxyeicosatetraenoic, 16-hydroxyeicosatetraenoic, 9-hydroxyoctadecadieneoic and 12-hydroxyeicosatetraenoic. No significant differences between AAD and LAD related to embryo quality were found.Conclusions/SignificanceOur study demonstrates for the first time that elevated concentrations of AAD and LAD in FF at the time of oocyte retrieval significantly decrease the ability of oocytes to form pronuclei after ICSI. This may serve as a new tool for non-invasive assessment of oocyte developmental capacity. However, levels of AAD and LAD are not associated with subsequent embryo quality or pregnancy rate, and therefore more studies are needed to determine their usefulness in human IVF procedure.
Highlights
There are many known techniques available to evaluate embryo quality [1,2,3,4], in vitro fertilization (IVF) needs reliable biomarkers of oocyte and sperm competence [4,5,6,7,8]
We have found significantly increased concentrations of hydroxyoctadecadienoic acid (HODE) and HETEs in the follicular fluid (FF) of oocytes that after intracytoplasmic sperm injection (ICSI) were unable to form pronuclei or underwent degeneration in comparison to those which achieved fertilization
Obtained results showed that arachidonic acid derivatives (AAD) and linoleic acid derivatives (LAD) correlated with the oocyte competence, not all derivatives correlated with zygote quality and neither AAD nor LAD demonstrated a relationship with embryo quality
Summary
There are many known techniques available to evaluate embryo quality [1,2,3,4], in vitro fertilization (IVF) needs reliable biomarkers of oocyte and sperm competence [4,5,6,7,8]. To avoid invasive techniques that could damage oocyte structure, a composition of follicular fluid (FF) has been investigated as a possible source of follicle biomarkers for oocyte quality [12, 13]. The COX enzyme pathway has recently been given considerable attention [17, 18], little information is available on the possible importance of the LOX and CYP pathways [19, 20]
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