Sá Barretto Letícia Siqueira, Castro Viviane Sgobbi Dias Caiado, Garcia Joaquim Mansano, Mingoti Gisele Zoccal
School of Agricultural and Veterinary Sciences, Department of Animal Reproduction, UNESP, 14884-900 Jaboticabal, SP, Brazil.
Zygote. 2011 May;19(2):107-16. doi: 10.1017/S096719941000016X. Epub 2010 Jun 23.
Aiming to improve the developmental competence of bovine oocytes during meiotic block, this study evaluated the effects of a serum replacer (Knockout SR®) and hormones (gonadotropins and estradiol) supplementation of prematuration medium (TCM119 with 0.5 mM IBMX [IBMX group] or 25 μM roscovitine [ROSC group]) on the kinetics of oocyte nuclear maturation and embryo development. Most IBMX and ROSC oocytes prematured for 8 h culture remained in the GV stage (70.3% and 73.1%, respectively; p > 0.05) similar to Control 8 h (63.5%) and to control immature oocytes (Control 0 h, 92.5%). After prematuration for 16 h, no oocytes remained in the GV stage at similar rates to those recently aspirated (p < 0.05); GV rates in ROSC (32.4%) were higher (p < 0.05) than in the Control 16 h group (8.6%), but similar (p > 0.05) to IBMX (9.7%). After in vitro maturation (IMV) for 24 h, metaphase II (MII) rates for oocytes prematured during 8 h were similar (p > 0.05) between control and treatments (65.0-71.7%). Similarly, MII rates oocytes prematured during 16 h were similar (p > 0.05) between all groups (45.9-60.4%). Cleavage rates (67.8-78.2%), embryonic development in day-7 (25.0-35.6%) and hatching rates in day-8 (2.5-11.3%) oocytes blocked during 8 h were similar for all groups (p > 0.05). Results indicate that addition of Knockout SR® and hormones to meiotic block culture with IBMX and roscovitine negatively affected meiotic arrest, but did not impair oocyte nuclear maturation and acquisition of developmental competence.
为提高牛卵母细胞减数分裂阻滞期间的发育能力,本研究评估了在早熟培养基(含0.5 mM异丁基甲基黄嘌呤的TCM119 [IBMX组]或含25 μM罗斯考维汀的TCM119 [ROSC组])中添加血清替代物(Knockout SR®)和激素(促性腺激素和雌二醇)对卵母细胞核成熟动力学及胚胎发育的影响。大多数在8小时培养中早熟的IBMX和ROSC卵母细胞仍处于生发泡(GV)期(分别为70.3%和73.1%;p>0.05),与对照8小时组(63.5%)和未成熟对照卵母细胞(对照0小时,92.5%)相似。早熟16小时后,没有卵母细胞以与刚采集时相似的比例停留在GV期(p<0.05);ROSC组的GV期比例(32.4%)高于对照16小时组(8.6%,p<0.05),但与IBMX组(9.7%)相似(p>0.05)。体外成熟(IVM)24小时后,8小时早熟卵母细胞的中期II(MII)比例在对照和各处理组之间相似(p>0.05)(65.0 - 71.7%)。同样,16小时早熟卵母细胞的MII比例在所有组之间相似(p>0.05)(45.9 - 60.4%)。所有组8小时阻滞的卵母细胞的卵裂率(67.8 - 78.2%)、第7天的胚胎发育率(25.0 - 35.6%)和第8天的孵化率(2.5 - 11.3%)相似(p>0.05)。结果表明,在含IBMX和罗斯考维汀的减数分裂阻滞培养中添加Knockout SR®和激素对减数分裂阻滞有负面影响,但不损害卵母细胞核成熟和发育能力的获得。