Expression of leptin ligand and receptor and effect of exogenous leptin supplement on in vitro development of porcine embryos.
The present study investigated the expression of ligand and receptor for leptin, and the effect of leptin supplementation on preimplantation development of porcine in vitro fertilized (IVF) and somatic cell nuclear transfer (SCNT) embryos. The IVF embryos were produced using frozen boar semen and SCNT embryos were obtained by nuclear transfer of fetal fibroblasts into enucleated oocytes. The protein expression of leptin ligand and receptor was investigated in in vitro matured oocytes, 2-, 4- and 8-cell embryos, morulae and blastocysts derived from IVF and SCNT using immunofluorescence. Both the ligand and receptor were detected in in vitro matured oocytes and all stage of IVF and SCNT embryos. The IVF and SCNT embryos were cultured in modified North Carolina State University (mNCSU)-23 medium supplemented with various concentrations (0, 1, 10, 100 or 1000 ng/mL) of leptin. The rates of cleavage at day 2 and blastocyst formation at day 7, and cell number of blastocysts were monitored as experimental parameters. In SCNT embryos, supplementing with 1000 ng/mL leptin significantly (P<0.05) increased the rate of blastocysts formation (20.2% versus 12.9%) and total cell number (54.6+/-17.4 versus 45.1+/-15.2) compared to the control group. In IVF embryos, leptin supplementation did not affect preimplantation embryo development and cell number in blastocysts. In conclusion, the present study demonstrated the expression of leptin ligand and receptor and the embryotropic effect of leptin in SCNT 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
Kim
Ji-hye
JH
Kang
Sung-keun
SK
Lee
Byeong-chun
BC
Hwang
Woo-suk
WS
D000818:Animals; D001755:Blastocyst / Q000737:chemistry / Q000166:cytology; D002452:Cell Count; D046149:Embryo Culture Techniques; D047108:Embryonic Development / Q000187:drug effects*; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary; D020738:Leptin / Q000032:analysis* / Q000494:pharmacology*; D009028:Morula / Q000737:chemistry; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000737:chemistry / Q000648:ultrastructure; D011956:Receptors, Cell Surface / Q000032:analysis*; D054411:Receptors, Leptin; D013552:Swine / Q000196:embryology*
D020738:Leptin; D011956:Receptors, Cell Surface; D054411:Receptors, Leptin