| Autor: |
Kim, Keun Jung, Lee, Ji Hye, Li, Xiaoxia, Kim, Eun Young, Park, Youn Bae, Park, Kang Sun, Ha, Yu Na, Kim, Maria, Han, Kil Woo, Yu, Jeong, Lee, Dong Soo, Lim, Jae Sam, Baek, Jun Jong, Kim, Jong Sang, Kim, Min Kyu |
| Zdroj: |
Biology of Reproduction; August 2012, Vol. 87 Issue: 1, Number 1 Supplement 1 p212-212, 1p |
| Abstrakt: |
The developmental competence of porcine IVM oocytes has unstable; because the cytoplasmic maturation was especially incomplete. For that reason, many report suggested supplementation of chemicals or some substances into IVM medium to improve maturation efficiency of oocytes. Acteoside is a typical phenylethanoid glycoside, extracted from various plants. It has various biological functions such as anti-oxidant, anti-inflammation, and anti-hypertension and been used pharmaceutically. Specially, it was powerful anti-oxidants either by direct scavenging of reactive oxygen and nitrogen species, or by acting as chain-breaking peroxyl radical scavengers. The aims of the present studies were to investigate that effect of acteoside supplemented to IVM medium on meiotic maturation and subsequent development of porcine oocytes. We examined the role of acteoside in IVM medium on the morphological progress of meiosis, developmental competence (Exp. 1). And the level of ROS in each oocyte was measured by ROS detection kit (Exp. 2). And we investigated effect of acteoside on the cytoplasmic maturation by homogeneous distribution and formation of cytoplasmic organelles by transmission electron microscopy (Exp. 3). And genes of apoptosis-regulation were selected for the RT-PCR, including anti-apoptotic genes such as Mcl-1, Bcl-2, and Bcl-xL and pro-apoptotic genes such as Bax, and Bak (Exp. 4) in the blastocyst after pathenogenetic activation. Oocytes were matured in tissue culture medium-199, supplemented with acteoside at various concentrations: 0 (control) 10, 30 and 50 μM. And electrical activation was executed according to direct-current pulse of 1.8 kV/cm for 30 μs was applied single pulse, using a BTX electro-Cell Manipulator 2001. The result of Exp. 1, oocytes maturation rates of groups supplemented with 10, 30, 50 μM acteoside (75.96, 72.95 and 73.68%, respectively) were no significantly different compared with control group (71.13%). And the number of parthenogenetic blastocysts was significantly higher in 10 μM acteoside treated group compared to control group (40.03 vs. 22.95%). In Exp. 2, the level of ROS (intensity/embryos) was significantly decreased in acteoside treated group (1.13, 0.95 and 1.35, respectively) compared with control group (6.38). In Exp. 3, the mitochondria and lipid droplet were smaller and homogeneous distribution in cytoplasm in the presence of 10 μM acteoside. And in Exp. 4, reverse transcription polymerase chain reaction (RT-PCR) of parthenogenetic blstocysts revealed that acteoside increased the anti-apoptotic genes, otherwise reduced pro-apoptotic genes. In conclusion, acteoside supplement was able to improve cytoplasmic maturation by not only homogeneous distribution and formation of cytoplasmic organelles but also regulation of pro-, anti-apoptotic gene. Our results represents that addition of acteoside to the IVM medium has a beneficial effect in physiology of porcine oocytes such as viability and activation, providing a improved method for porcine oocytes in vitro. This work was supported by a grant (Code# PJ008148) from BioGreen21 Program, Rural Development Administration, Republic of Korea. |
| Databáze: |
Supplemental Index |
| Externí odkaz: |
|
