58 EFFECT OF LYSOPHOSPHATIDIC ACID ON PORCINE OOCYTE IN VITRO MATURATION AND SUBSEQUENT EMBRYONIC DEVELOPMENT AFTER PARTHENOGENETIC ACTIVATION AND IVF

Abstract

Lysophosphatidic acid (LPA) is a signalling molecule derived from phospholipid, known to have biological activities such as stimulating cell proliferation, differentiation, and migration. The purpose of this study was to investigate the effect of LPA on in vitro maturation (IVM) of porcine oocytes and subsequent embryonic development after in vitro fertilization (IVF) and parthenogenetic activation (PA). We examined nuclear maturation, intracellular glutathione (GSH), and reactive oxygen species (ROS) levels and subsequent embryonic development after IVF in porcine oocytes matured in TCM-199 media containing either 0, 10, 30, or 60 μM LPA (No. of cultured oocytes 225, 225, 230, and 226, respectively). All data were compared for all groups using one-way ANOVA, followed by Duncan’s multiple range test. After 44 h of IVM, no significant difference was observed for nuclear maturation from the different (0, 10, 30, and 60 μM) LPA treatment groups (86.61, 87.48, 92.97, and 88.69%, respectively) compared with the control (P < 0.05). The 30 μM LPA treated group exhibited a significant (P < 0.05) increase in intracellular GSH levels and decrease in intracellular ROS levels compared with other LPA-treated groups. Oocytes matured with 30 μM LPA during IVM had significantly (P < 0.05) higher cleavage rates after PA (88.50%) than other LPA-treated groups (79.42, 83.87, and 83.65%, respectively). Blastocyst formation rates following PA increased up to 30 μM LPA, but the rates decreased in the 60 μM LPA-treated group (49.64, 53.28, 57.00, and 51.09%, respectively). However, a statistical difference was not observed (P < 0.05). Upon evaluation of embryonic competence following IVF, treatment with 30 μM LPA during IVM significantly increased cleavage rates (70.44%; P < 0.05) compared with 0 and 60 μM LPA-treated groups (61.51 and 65.27%, respectively). Blastocyst formation rates improved with increasing concentrations of LPA up to 30 µM; however, the rates decreased in the 60 μM LPA-treated group (26.34, 30.10, 33.26, and 27.77%, respectively). However, like the preceding PA blastocyst formation rates, a statistical difference was not observed (P < 0.05). In conclusion, treatment with 30 μM LPA during IVM improved the developmental potential of PA and IVF porcine embryos by increasing the intracellular GSH level, thereby decreasing the intracellular ROS level during oocyte maturation.

This work was supported, in part, by a grant from the Cooperative Research Program for Agriculture Science & Technology Development (Project No. PJ011077, PJ011288), Rural Development Administration, and the National Research Foundation of Korea Grant funded by the Korean Government (NRF-2013R1A2A2A04008751). These are the results of a study from the Leaders INdustry-University Cooperation Project 2015-B-0013–010118, supported by the Ministry of Education, Republic of Korea.