Caffeine treatment prevents age-related changes in ovine oocytes and increases cell numbers in blastocysts produced by somatic cell nuclear transfer.
Maturation-promoting factor (MPF) and mitogen-activated protein kinase (MAPK) are key regulators of both meiotic and mitotic cycles. Oocytes arrested at metaphase of the second meiotic division (MII) contain high levels of both kinases; however, these activities decline with age. Caffeine (an inhibitor of Myt1/Wee1 activity) can increase MPF and MAPK activities in ovine oocytes; however, the effects of caffeine treatment on the activation, nuclear configuration and developmental potential of ovine SC nuclear transfer (SCNT) embryos were unknown. We examined the effects of aging and caffeine treatment on MPF and MAPK activities, activation, development, and nuclear remodeling of SCNT embryos. Both kinases reached maximum activities at 24-h postonset of maturation (hpm) and then decreased with time. The decline in MPF activity occurred rapidly, whereas MAPK activity declined more slowly. Caffeine treatment (10.0 mM) of aging oocytes prevented the decline in activities associated with both kinases and prevented the acquisition of activation competence by a single activation stimulus. However, treatment of aged oocytes with caffeine could not increase kinase activities or reverse the acquisition of activation competence. Enucleation did not affect kinase activities, but caffeine treatment significantly increased both. Caffeine treatment did not affect the decline in MPF or MAPK activities following activation or significantly affect development of parthenogenetically activated oocytes. When SCNT reconstructed embryos were treated with caffeine following fusion, no increase in the frequency of development to blastocyst was observed; however, a significant increase in the occurrence of nuclear envelope break-down (NEBD) and an increase in total cell numbers occurred.
Lee
Joon-Hee
JH
Animal Development and Biotechnology Group, Division of Animal Physiology, School of Biosciences, The University of Nottingham, Sutton Bonington, Loughborough, Leicestershire, United Kingdom.
Campbell
Keith H S
KH
D000367:Age Factors; D000818:Animals; D001755:Blastocyst / Q000166:cytology* / Q000187:drug effects* / Q000502:physiology; D002110:Caffeine / Q000494:pharmacology*; D000001:Calcimycin / Q000378:metabolism; D007476:Ionophores / Q000378:metabolism; D016200:Maturation-Promoting Factor / Q000378:metabolism; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000166:cytology / Q000187:drug effects* / Q000502:physiology*; D010312:Parthenogenesis; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology*; D012756:Sheep
D007476:Ionophores; D010726:Phosphodiesterase Inhibitors; D000001:Calcimycin; D002110:Caffeine; D016200:Maturation-Promoting Factor; D020928:Mitogen-Activated Protein Kinases