Caffeine promotes premature chromosome condensation formation and in vitro development in porcine reconstructed embryos via a high level of maturation promoting factor activity during nuclear transfer.
When the nucleus in G0/G1 phase is transferred to an enucleated oocyte by nuclear transfer (NT), its nuclear envelope is broken, followed by condensation of chromosome structure by maturation promoting factor (MPF). This morphological remodeling of the transferred interphase nucleus seems to be essential for subsequent development of NT embryos. In this study, we treated porcine NT embryos with caffeine, which has been reported to increase MPF activity, to keep their MPF level high during NT. When 2.5 mM caffeine was added to the handling medium, the proportion of NT embryos showing condensed chromosome increased significantly (P < 0.05). In NT embryos treated with caffeine, the activity of p34(cdc2) kinase was significantly (P < 0.05) higher than in those without caffeine at 3 h post-injection. In addition, the rate of development to the blastocyst stage after activation was significantly (P < 0.05) higher in NT embryos treated with caffeine. These results indicate that caffeine treatment can increase not only the rate of chromosome condensation but also the developmental rate to the blastocyst stage of porcine NT embryos. This action is most likely due to the support/increase of MPF activity throughout the process of NT.
Kawahara
Manabu
M
Laboratory of Animal Reproduction, Graduate School of Agricultural Science, Tohoku University, Sendai 981-8555, Japan. mana1976@nodai.ac.jp
Wakai
Takuya
T
Yamanaka
Ken-Ichi
K
Kobayashi
Jin
J
Sugimura
Satoshi
S
Shimizu
Takashi
T
Matsumoto
Hiromichi
H
Kim
Jin-Hoi
JH
Sasada
Hiroshi
H
Sato
Eimei
E
D000818:Animals; D002110:Caffeine / Q000494:pharmacology*; D042002:Chromatin Assembly and Disassembly; D019976:Cloning, Organism; D046149:Embryo Culture Techniques; D004622:Embryo, Mammalian / Q000378:metabolism; D047108:Embryonic Development / Q000187:drug effects*; D005260:Female; D016200:Maturation-Promoting Factor / Q000494:pharmacology*; D053652:Nuclear Transfer Techniques; D011247:Pregnancy; D013268:Stimulation, Chemical; D013552:Swine
D002110:Caffeine; D016200:Maturation-Promoting Factor