Embryotropic effect of glycosaminoglycans and receptors in development of porcine pre-implantation embryos.
The present study investigated the expression of receptors for glycosaminoglycans (GAGs) and the effect of GAGs supplementation on development of porcine IVF embryos. Total RNA was prepared from oocytes, 2-, 4- and 8-cell embryos, morulae and blastocysts. The expression of hyaluronic acid receptor (CD44) and heparin (HP) interacting protein (HIP) was determined using RT-PCR and Southern blot analysis. The CD44 and HIP mRNA were detected from in vitro matured oocytes and all stages of pre-implantation embryos. The IVF embryos were cultured in modified NCSU-23 medium supplemented with various concentrations (0, 0.1, 0.5 or 1.0 mg/mL) of hyaluronic acid (HA) or heparin. Supplementing with 0.5 mg/mL HA significantly increased total cell number compared to other experimental groups, due to increase in trophectoderm cells. Supplementing with 1.0 mg/mL, HP significantly increased blastocyst formation rate compared to the control group. Supplementing media, in which IVF embryos were cultured with 0.5 mg/mL HA + 1.0 mg/mL HP, significantly increased blastocyst formation rate compared to the control group. In conclusion, the present study demonstrated the expression of HA and HP receptors and the embryotrophic effect of HA or HP on porcine IVF embryos.
Kim
Hye-Soo
HS
Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, Seoul 151-742, South Korea.
Lee
Gab-Sang
GS
Hyun
Sang-Hwan
SH
Nam
Dong-Hyun
DH
Lee
So-Hyun
SH
Jeong
Yeon-Woo
YW
Kim
Sue
S
Kim
Ji-Hye
JH
Kang
Sung-Keun
SK
Lee
Byeong-Chun
BC
Hwang
Woo-Suk
WS
D000818:Animals; D001755:Blastocyst; D015139:Blotting, Southern; D003470:Culture Media; D046149:Embryo Culture Techniques / Q000662:veterinary; D047108:Embryonic Development / Q000187:drug effects*; D015870:Gene Expression; D006025:Glycosaminoglycans / Q000494:pharmacology*; D006493:Heparin / Q000008:administration & dosage; D018960:Hyaluronan Receptors / Q000235:genetics; D006820:Hyaluronic Acid / Q000008:administration & dosage; D012333:RNA, Messenger / Q000032:analysis; D011956:Receptors, Cell Surface / Q000235:genetics / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013552:Swine / Q000196:embryology*
D003470:Culture Media; D006025:Glycosaminoglycans; D018960:Hyaluronan Receptors; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; C086020:glycosaminoglycan receptor; C037680:heparin receptor; D006820:Hyaluronic Acid; D006493:Heparin