Bagg Melanie A, Nottle Mark B, Grupen Christopher G, Armstrong David T
Department of Obstetrics and Gynaecology, Research Centre for Reproductive Health, University of Adelaide, The Queen Elizabeth Hospital, Woodville, South Australia, Australia.
Mol Reprod Dev. 2006 Oct;73(10):1326-32. doi: 10.1002/mrd.20555.
Pre-pubertal pig oocytes display reduced developmental competence compared with adult oocytes following in vitro maturation (IVM). Exposure to dibutyryl cyclic adenosine monophosphate (dbcAMP) for the first 20 hr IVM improves development of pre-pubertal oocytes, suggesting that their cAMP content may be inadequate. This study examined the effect of 1 mM dbcAMP treatment for the first 22 hr of IVM on the cAMP content, meiotic progression, and embryo development of pre-pubertal and adult oocytes. In control groups, a two-fold increase in cAMP was observed in adult oocytes after 22 hr IVM, with no change in pre-pubertal oocyte cAMP content. At 22 hr IVM, dbcAMP treatment resulted in two- and five-fold increases in pre-pubertal and adult oocyte cAMP, respectively. After 22 hr control IVM, a greater proportion of pre-pubertal oocytes occupied metaphase I (MI) compared with adult oocytes (69% vs. 49%). dbcAMP treatment reduced the proportion of pre-pubertal and adult oocytes in MI stage at 22 hr. Despite dbcAMP treatment, the proportion of pre-pubertal oocytes in the MI stage at 22 hr remained higher than that of adult oocytes. In control groups, adult oocytes displayed a greater ability to form blastocysts compared with pre-pubertal oocytes following either parthenogenetic activation (59% vs. 25%) or in vitro fertilization (IVF) (47% vs. 19%). dbcAMP treatment increased subsequent blastocyst formation rates of pre-pubertal oocytes, whereas blastocyst formation rates of adult oocytes remained unchanged. Our results suggest that the reduced developmental capacity of pre-pubertal oocytes may be a consequence of their reduced ability to accumulate cAMP during IVM.
与体外成熟(IVM)后的成年猪卵母细胞相比,青春期前猪卵母细胞的发育能力较低。在IVM的前20小时暴露于二丁酰环磷酸腺苷(dbcAMP)可改善青春期前卵母细胞的发育,这表明它们的cAMP含量可能不足。本研究检测了在IVM的前22小时用1 mM dbcAMP处理对青春期前和成年猪卵母细胞的cAMP含量、减数分裂进程及胚胎发育的影响。在对照组中,IVM 22小时后成年卵母细胞中的cAMP增加了两倍,而青春期前卵母细胞的cAMP含量没有变化。在IVM 22小时时,dbcAMP处理使青春期前和成年卵母细胞的cAMP分别增加了两倍和五倍。在IVM 22小时的对照处理后,与成年卵母细胞相比,处于中期I(MI)的青春期前卵母细胞比例更高(69%对49%)。dbcAMP处理降低了22小时时处于MI期的青春期前和成年卵母细胞的比例。尽管进行了dbcAMP处理,但22小时时处于MI期的青春期前卵母细胞比例仍高于成年卵母细胞。在对照组中,孤雌激活(59%对25%)或体外受精(IVF)(47%对19%)后,成年卵母细胞形成囊胚的能力高于青春期前卵母细胞。dbcAMP处理提高了青春期前卵母细胞随后的囊胚形成率,而成年卵母细胞的囊胚形成率保持不变。我们的结果表明,青春期前卵母细胞发育能力降低可能是其在IVM期间积累cAMP能力下降的结果。