Abstract
Traditional methods for the evaluation of oocyte quality are based on morphological classification of the follicle, cumulus-oocyte complex, polar body and meiotic spindle. This study is focused on the differences between the morphological assessment of oocyte quality, the assessment based on Lissamine Green B (LB) staining and the analysis of oocytes using a proteomic approach. We evaluated the effectiveness of electrochemical and chemical parthenogenetic activation under our laboratory conditions and evaluated the applicability of Lissamine Green B staining of cumulus-oocyte complexes (COCs) as a non-invasive method for predicting the maturational and developmental competence of porcine oocytes cultured in vitro. We determined that chemical parthenogenetic activation using ionomycin and 6-dimethylaminopurine was slightly more effective than electrochemical activation. After oocyte selection according to LB staining, we found significant differences (P<0.05) between the LB- group and LB+ group and the control group in their maturation, cleavage rate and rate of blastocysts. Proteomic analyses identified a selection of proteins that were differentially expressed in each group of analysed oocytes. Oocytes of the LB- group exhibited an increased variability of proteins involved in transcription regulation, proteosynthesis and the protein folding crucial for oocyte maturation and further embryonic development. These results found a better competence of LB- oocytes in maturation, cleavage and ability to reach the blastocyst stage.
Highlights
Oocytes quality has a significant influence on the success of in vitro maturation (IVM), fertilization (IVF), embryo development and possible implantation (Goto et al, 1988; Sirard et al, 2006; Hoshino, 2018; Murin et al, 2019)
The cumulus-oocyte complexes (COCs) obtained from the local slaughterhouse were divided into several experimental groups and used for our experiments, which based on the comparison of different techniques of parthenogenetic activation, Lissamine Green B (LB) staining and proteomic profiling
In total 268 COCs were divided into 3 experimental groups: the control group (CG), LB+ group and LB- group
Summary
Oocytes quality has a significant influence on the success of in vitro maturation (IVM), fertilization (IVF), embryo development and possible implantation (Goto et al, 1988; Sirard et al, 2006; Hoshino, 2018; Murin et al, 2019). Lower oocyte quality and occurrence of morphological anomalies are related a lower rate of IVM, IVF and IVC, which means, that oocyte quality is a key attribute of effective in vitro manipulations and treatment of infertility. This fact is a reason why oocyte collection is based on a strictly qualitative assessment of the current state of oocytes and embryos (Vassena et al, 2003; Krisher, 2004).
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