Osmolarity at early culture stage affects development and expression of apoptosis related genes (Bax-alpha and Bcl-xl) in pre-implantation porcine NT embryos.
This study examined whether high osmolarity of culture medium at the early culture stage affects development and expression of apoptosis related genes (Bax-alpha and Bcl-xl) of porcine nuclear transfer (NT) and in vitro fertilization (IVF) embryos. NT and IVF embryos were divided into three groups and the basic medium was PZM-3 (260-270 mOsmol, control group). The control group of embryos was cultured in PZM-3 for whole culture period. Other two groups of embryos were cultured in a modified PZM-3 with 0.05 M sucrose (300-320 mOsmol, sucrose group) or increased NaCl to 138 mM (300-320 mOsmol, NaCl group) for the first 2 days, and then cultured in PZM-3 for 4 days. NT embryos cultured in NaCl group showed a significantly higher developmental rate to the blastocyst stage with a decreased apoptosis rate compared to the control (P < 0.05). There was no difference in blastocyst formation and apoptosis incidence among the three culture treatments for IVF-derived embryos. Bax-alpha mRNA expression was significantly higher in the control than sucrose or NaCl group for both NT and IVF embryos (P < 0.05). Moreover, the relative abundance of Bax-alpha/Bcl-xl was higher in the control than the treatment groups. These results indicate that the higher osmolarity at the early embryonic stage of porcine NT and IVF embryos can improve the in vitro development with reduced apoptosis through regulating the Bax-alpha/Bcl-xl gene expression.
Hwang
In-Sun
IS
Division of Animal Biotechnology, National Livestock Research Institute, RDA, Suwon, Korea.
Park
Mi-Rung
MR
Moon
Hyo-Jin
HJ
Shim
Joo-Hyun
JH
Kim
Dong-Hoon
DH
Yang
Byoung-Chul
BC
Ko
Yeoung-Gyu
YG
Yang
Boh-Suk
BS
Cheong
Hee-Tae
HT
Im
Gi-Sun
GS
D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D001755:Blastocyst / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D002478:Cells, Cultured; D003470:Culture Media / Q000494:pharmacology*; D046149:Embryo Culture Techniques; D004622:Embryo, Mammalian; D047108:Embryonic Development / Q000187:drug effects* / Q000235:genetics; D005307:Fertilization in Vitro; D018507:Gene Expression Regulation, Developmental; D009994:Osmolar Concentration; D013552:Swine / Q000196:embryology*; D013997:Time Factors; D014882:Water-Electrolyte Balance / Q000502:physiology; D051028:bcl-2-Associated X Protein / Q000235:genetics* / Q000378:metabolism; D051020:bcl-X Protein / Q000235:genetics* / Q000378:metabolism
D003470:Culture Media; D051028:bcl-2-Associated X Protein; D051020:bcl-X Protein