JARB Journal of Animal Reproduction and Biotehnology

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Journal of Embryo Transfer 2012; 27(3): 163-169

Published online September 30, 2012

Copyright © The Korean Society of Animal Reproduction and Biotechnology.

The Effects of 3-Isobutyl-1-methylxanthine (IBMX) on Nuclear and Cytoplasmic Maturation of Porcine Oocytes In Vitro

Kwak, Seong-Sung, Jang, Seung-Hoon, Jeong, Se-Heon, Jeon, Yubyeol, Biswas, Dibyendu, Hyun, Sang-Hwan

Laboratory of Veterinary Embryology and Biotechnology, Department of Veterinary Medicine, College of Veterinary Medicine, Chungbuk National University, Cheongju 361-763, Korea

Abstract

The 3-isobutyl-1-methylxanthine (IBMX) is non-selective phosphodiesterase and is able to prevent resumption of meiosis by maintaining elevated cyclic AMP (cAMP) concentrations in the oocyte. The present study was conducted to analyze: (1) nuclear maturation (examined by the Hoechst staining), (2) whether cytoplasmic maturation (examined by the intracellular glutathione (GSH) concentration) of porcine oocytes is improved during meiotic arrest after prematuration (22 h) with IBMX. Before in vitro maturation (IVM), oocytes were treated with 1 mM IBMX for 22 h. After 22 h of pre-maturation, the higher rate of IBMX treated group oocytes were arrested at the germinal vesicle (GV) stage (42.3%) than control IVM oocytes (10.1%). It appears that the effect of IBMX on the resumption of meiosis has shown clearly. In the end of IVM, the reversibility of the IBMX effect on the nuclear maturation has been corroborated in this study by the high proportions of MII stage oocytes (72.5%) reached after 44 h of IVM following the 22 h of inhibition. However, intracellular GSH concentrations were lower in the oocytes treated with IBMX than the control oocytes (6.78 and 12.94 pmol/oocyte, respectively). These results demonstrate that cytoplasmic maturation in porcine oocytes pre-treated with IBMX for 22 h did not equal that of control oocytes in the current IVM system. These results indicate that pre-maturation with IBMX for 22 h may not be beneficial in porcine IVM system.

Keywords: porcine oocytes, IBMX, phosphodiesterase, cAMP, resumption of meiosis, nuclear maturation