Short-term treatment with 6-DMAP and demecolcine improves developmental competence of electrically or Thi/DTT-activated porcine parthenogenetic embryos.
Treatment with 6-dimethylaminopurine (6-DMAP) or demecolcine (DE) for several (at least 2) hours after artificial activation is known to improve in vitro development of porcine embryos. However, several reports have also shown that treatments with these chemicals induce apoptosis. The aim of this study was to find out whether short-term treatment with 6-DMAP and DE combined with electrical or thimerosal/dithiothreitol (Thi/DTT) activation had a beneficial effect on development of parthenogenetically activated porcine oocytes. We additionally treated embryos with 6-DMAP (2 mM) and/or DE (0.4 microg/ml) for a short time (40 min) after an electrical pulse (EP) or Thi/DTT. As a result, short-term treatment with 6-DMAP and DE successfully induced development of electrically or Thi/DTT-activated porcine parthenogenetic embryos with no significant difference in cleavage rate, blastocyst formation rate and total cell number compared with long-term treatment. To find optimal activation protocol, cleavage rate, blastocyst formation rate and total cell number were compared between EP and Thi/DTT treatments. Thi/DTT + 6-DMAP + DE showed significantly higher blastocyst formation rate (36.1 ± 3.5%) and total cell number (46.9 ± 1.0) than other groups (EP + 6-DMAP + DE, EP + Thi/DTT + 6-DMAP + DE: 23.3 ± 3.0%, 42.2 ± 1.1 and 17.2 ± 2.7%, 36.7 ± 1.5, respectively). In conclusion, this study demonstrates that short-term treatment with 6-DMAP and DE is as effective as the standard long-term treatment and Thi/DTT + 6-DMAP + DE exerts a synergistic effect.
Park
Sol Ji
SJ
Department of Theriogenology and Biotechnology, College of Veterinary Medicine and Research Institute for Veterinary Science, Seoul National University, Seoul, 151-742, Korea.
Koo
Ok Jae
OJ
Kwon
Dae Kee
DK
Gomez
Ma Ninia Limas
MN
Kang
Jung Taek
JT
Atikuzzaman
Mohammad
M
Kim
Su Jin
SJ
Jang
Goo
G
Lee
Byeong Chun
BC
D000225:Adenine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000187:drug effects; D002467:Cell Nucleus / Q000502:physiology; D003703:Demecolcine / Q000494:pharmacology*; D004229:Dithiothreitol / Q000494:pharmacology*; D004558:Electric Stimulation; D004622:Embryo, Mammalian / Q000187:drug effects* / Q000378:metabolism; D047108:Embryonic Development / Q000187:drug effects*; D005260:Female; D009865:Oocytes / Q000187:drug effects / Q000502:physiology; D010312:Parthenogenesis / Q000187:drug effects; D013552:Swine; D013849:Thimerosal / Q000494:pharmacology*
D013849:Thimerosal; C001020:N(6),N(6)-dimethyladenine; D000225:Adenine; D004229:Dithiothreitol; D003703:Demecolcine