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当前位置: 首页 > 文章 > Effects of LPA on the development of sheep in vitro fertilized embryos and attempt to establish sheep embryonic stem cells 农业科学学报 (英文) 2023,22 (4)
Position: Home > Articles > Effects of LPA on the development of sheep in vitro fertilized embryos and attempt to establish sheep embryonic stem cells Journal of Integrative Agriculture 2023,22 (4)

Effects of LPA on the development of sheep in vitro fertilized embryos and attempt to establish sheep embryonic stem cells

作  者：
ZHANG, Xue-min;HUANG, Xiang-hua;WANG, Jing;XING, Ying;LIU, Fang;XIANG, Jin-zhu;WANG, Han-ning;YUE, Yong-li;LI, Xue-ling
单  位：
Inner Mongolia Univ, Sch Life Sci, State Key Lab Reprod Regulat & Breeding Grassland, Hohhot 010020, Peoples R China;Inner Mongolia Autonomous Reg Peoples Hosp, Dept Urol, Hohhot 010017, Peoples R China
关键词：
sheep;in vitro fertilization;LPA;LPARs;embryonic stem cells
摘  要：
Lysophosphatidic acid (LPA) is a small molecule glycerophospholipid, which regulates multiple downstream signalling pathways through G-protein-coupled receptors to achieve numerous functions on oocyte maturation and embryo development. In this study, sheep in vitro fertilized embryos were applied to investigate the effects of LPA on early embryos development and embryonic stem cell establishment. At first, the maturation medium containing estrus female sheep serum and synthetic oviduct fluid (SOF) were optimized for sheep IVF, and then the effects of LPA were investigated. From 0.1 to 10 pmol L-1, LPA had no significant effect on the cleavage rate (P>0.05), but the maturation rate and blastocyst rate increased dependently with LPA concentration (P<0. 05), and the blastocyst morphology was normal. When the LPA concentration was 15 pmol L-1, the maturation rate, cleavage rate and blastocyst rate decreased significantly (P<0.05), and the blastocyst exhibited abnormal morphology and could not develop into high -quality blastocyst. Besides, the exogenous LPA increases the expression of LPAR2, LPAR4, TE-related gene CDX-2 and pluripotency-related gene OCT-4 in sheep early IVF embryos with the raise of LPA concentration from 0.1 to 10 pmol L-1. The expression of LPAR2, LPAR4, CDX-2 and OCT-4 from the LPA-0.1 pmol L-1 to LPA-10 pmol L-1 groups in early embryos were extremely significant (P<0.05), while the expression of these genes significantly decreased in 15 pmol L-1 LPA-treated embryos compared with LPA-10 pmol L-1 group (P<0.05). The inner cell mass in 15 pmol L-1 LPA-treated embryos was also disturbed, and the blastocysts formation was abnormal. Secondly, the sheep IVF blastocysts were applied to establish embryonic stem cells. The results showed that LPA made the blastocyst inoculated cells grow towards TSC-like cells. They enhanced the fluorescence intensity and mRNA abundance of OCT-4 and CDX-2 as the concentration increased from 0 to 10 pmol L-1, while 15 pmol L-1 LPA decreased OCT-4 and CDX-2 expression in the derived cells. The expression of CDX-2 and OCT-4 in the blastocyst inoculated cells of LPA-1 pmol L-1 group and LPA-10 pmol L-1 group extremely significantly increased (P<0.05), but there was significant decrease in LPA-15 mu mol L-1 group compared with LPA-10 mu mol L-1 group (P<0.05). Meanwhile, the protein expression of LPAR2 and LPAR4 remarkably increased after treatment of LPA at 10 mu mol L-1 concentration. This study references the IVF embryo production and embryonic stem cell research of domestic animals.

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