Male pronuclear formation in denuded porcine oocytes after in vitro maturation in the presence of cysteamine.
The present study was conducted to examine effects of cysteamine in culture medium on progression of meiosis, glutathione (GSH) content, kinase activities (histone H1 kinase and mitogen-activated protein kinase), and male pronuclear formation after in vitro insemination of cumulus-denuded oocytes (DOs) in the pig. DOs, obtained by mechanically removing cells from cumulus-oocyte complexes (COCs) with a small-bore pipette, were cultured for 45 h in TCM199 supplemented with sodium pyruvate, gonadotropins, estradiol, and 10% porcine follicular fluid, with or without cysteamine (150 microM). Maturation rates of DOs cultured with and without cysteamine were not different (60-70%) but were significantly lower than those of COCs (90-100%) (p < 0.05). GSH content of matured DOs cultured with cysteamine was significantly higher than that of DOs cultured without cysteamine (p < 0.05). Values for both types of kinase activity in matured DOs cultured with and without cysteamine were not different (p > 0.05). After in vitro insemination, DOs cultured with cysteamine showed significantly higher rates of male pronuclear formation (80.3 +/- 3.0%) than DOs cultured without cysteamine (16.4 +/- 0.5%) (p < 0.05). These results indicate that the addition of cysteamine to culture medium increased oocyte GSH content and promoted male pronuclear formation after sperm penetration of porcine DOs but had no effects on their maturation rates or kinase activities.
Yamauchi
N
N
Department of Animal Production, Tohoku National Agricultural Experiment Station, Morioka 020-0123, Japan.
Nagai
T
T
D000818:Animals; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism; D002467:Cell Nucleus / Q000648:ultrastructure*; D002478:Cells, Cultured; D003470:Culture Media; D003543:Cysteamine / Q000008:administration & dosage / Q000494:pharmacology*; D005260:Female; D005307:Fertilization in Vitro; D015571:Follicular Fluid; D005978:Glutathione / Q000378:metabolism; D008297:Male; D008540:Meiosis / Q000187:drug effects; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure*; D011494:Protein Kinases / Q000378:metabolism; D013094:Spermatozoa / Q000648:ultrastructure*; D013552:Swine / Q000502:physiology*
D003470:Culture Media; D003543:Cysteamine; D011494:Protein Kinases; C102333:histone H1 kinase; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D005978:Glutathione