Effect of D-Glucuronic Acid and N-acetyl-D-Glucosamine Treatment during In Vitro Maturation on Embryonic Development after Parthenogenesis and Somatic Cell Nuclear Transfer in Pigs.

Abstract

Simple SummaryHyaluronic acid, also known as hyaluronan, is essential for the expansion of cumulus cells, the maturation of oocytes, and further embryo development. This study aimed to examine the effects of treatment with glucuronic acid and N-acetyl-D-glucosamine, which are components of hyaluronic acid, during porcine oocyte in vitro maturation and embryonic development after parthenogenetic activation and somatic cell nuclear transfer. We measured the diameter of mature oocytes, the thickness of the perivitelline space, the intracellular reactive oxygen species level, and the expression of cumulus cell expansion genes and reactive oxygen species-related genes and examined the cortical granule reaction of oocytes after electrical activation. In conclusion, the addition of 0.05 mM glucuronic acid and 0.05 mM N-acetyl-D-glucosamine and during the initial 22 h of in vitro maturation in pig oocytes has beneficial effects on cumulus expansion, perivitelline space thickness, cytoplasmic maturation, reactive oxygen species level, cortical granule exocytosis, and early embryonic development after parthenogenesis and somatic cell nuclear transfer. Glucuronic acid and N-acetyl-D-glucosamine can be applied to in vitro production technology and can be used as ingredients to produce high-quality porcine blastocysts.This study aimed to examine the effects of treatment with glucuronic acid (GA) and N-acetyl-D-glucosamine (AG), which are components of hyaluronic acid (HA), during porcine oocyte in vitro maturation (IVM). We measured the diameter of the oocyte, the thickness of the perivitelline space (PVS), the reactive oxygen species (ROS) level, and the expression of cumulus cell expansion and ROS-related genes and examined the cortical granule (CG) reaction of oocytes. The addition of 0.05 mM GA and 0.05 mM AG during the first 22 h of oocyte IVM significantly increased oocyte diameter and PVS size compared with the control (non-treatment). The addition of GA and AG reduced the intra-oocyte ROS content and improved the CG of the oocyte. GA and AG treatment increased the expression of CD44 and CX43 in cumulus cells and PRDX1 and TXN2 in oocytes. In both the chemically defined and the complex medium (Medium-199 + porcine follicular fluid), oocytes derived from the GA and AG treatments presented significantly higher blastocyst rates than the control after parthenogenesis (PA) and somatic cell nuclear transfer (SCNT). In conclusion, the addition of GA and AG during IVM in pig oocytes has beneficial effects on oocyte IVM and early embryonic development after PA and SCNT.