Anguita Begoña, Jimenez-Macedo Ana R, Izquierdo Dolors, Mogas Teresa, Paramio Maria-Teresa
Departament de Ciència Animal I dels Aliments, Universitat Autònoma de Barcelona, 08193 Bellaterra, Barcelona, Spain.
Theriogenology. 2007 Feb;67(3):526-36. doi: 10.1016/j.theriogenology.2006.09.003. Epub 2006 Oct 2.
The aim of this study was to analyze the relationship between oocyte diameter, meiotic and embryo developmental competence and the expression of the catalytic subunit of MPF, the p34(cdc2), at mRNA, RNA and protein level, as well as its kinase activity, in prepubertal (1-2 months old) goat oocytes. MPF is the main meiotic regulator and a possible regulator of cytoplasmic maturation; therefore, it could be a key factor in understanding the differences between competent and incompetent oocytes. Oocytes were classified according to oocyte diameter in four categories: <110, 110-125, 125-135 and >135 microm and matured, fertilized and cultured in vitro. The p34(cdc2) was analyzed in oocytes at the time of collection (0 h) and after 27 h of IVM (27 h) in each of the oocyte diameter categories. The oocyte diameter was positively related to the percentage of oocytes at MII after IVM (0, 20.7, 58 and 78%, respectively) and the percentage of blastocysts obtained at 8 days postinsemination (0, 0, 1.95 and 12.5%, respectively). The expression of RNA and mRNA p34(cdc2) did not vary between oocyte diameters at 0 and 27h. Protein expression of p34(cdc2) increased in each oocyte category after 27 h of maturation. MPF activity among diameter groups did not vary at 0h but after IVM there was a clear and statistically significant increase of MPF activity in the biggest oocytes.
本研究的目的是分析青春期前(1 - 2月龄)山羊卵母细胞中卵母细胞直径、减数分裂及胚胎发育能力与促成熟因子(MPF)催化亚基p34(cdc2)在mRNA、RNA和蛋白质水平的表达及其激酶活性之间的关系。MPF是主要的减数分裂调节因子,也是细胞质成熟的潜在调节因子;因此,它可能是理解有发育能力和无发育能力卵母细胞差异的关键因素。根据卵母细胞直径将其分为四类:<110、110 - 125、125 - 135和>135微米,并进行体外成熟、受精和培养。在每个卵母细胞直径类别中,于采集时(0小时)和体外成熟培养27小时后(27小时)对卵母细胞中的p34(cdc2)进行分析。卵母细胞直径与体外成熟后处于MII期的卵母细胞百分比(分别为0、20.7、58和78%)以及授精后8天获得的囊胚百分比(分别为0、0、1.95和12.5%)呈正相关。在0小时和27小时时,不同直径的卵母细胞中RNA和mRNA p34(cdc2)的表达没有差异。成熟27小时后,每个卵母细胞类别中p34(cdc2)的蛋白质表达均增加。在0小时时,不同直径组之间的MPF活性没有差异,但体外成熟后,最大的卵母细胞中MPF活性有明显且具有统计学意义的增加。