Li Y P, Cheng G X, Gu Z, Tso J K
Shanghai Institute of Cell Biology, Academia Sinica.
Shi Yan Sheng Wu Xue Bao. 1993 Dec;26(4):399-409.
Experiments were designed to evaluate the ability of rabbit oviduct epithelial cells (ROEC) or ROEC conditioned medium to promote the development of rat eggs fertilized in Vivo or in Vitro. 61.73% of the eggs fertilized in Vitro and 73.33% of the eggs fertilized in Vivo cocultured with ROEC overcame the 2-cell block (Plate I, tables 1 and 2). Similarly, 67.99% of the eggs fertilized in Vitro cultured in ROEC conditioned medium developed over the 2-cell stage, and nearly half of them developed to morula and blastocysts stage (Table 3). By using 35S-methionine incorporation and autoradiography methods, several polypeptides, with molecular weight of 135 Kd, 68 Kd, 55 Kd, 51 Kd and 44 Kd respectively (Fig. 1), were excreted from rabbit oviduct epithelial cells and found in the ROEC conditioned medium. The possibility of entrance of the ROEC proteins into the developing embryo was tested by determining whether any of the secreted proteins bound to the zona pellucida. The results of iodination by using 125I-containing acylating agent labelling method showed that the 68 Kd and 55 Kd proteins were bound onto the zona pellucida of rat eggs co-cultured with ROEC in vitro for 24 h (Fig. 2). Studies concerning the problem that whether these two secreted proteins are the key factors to promote the development of early embryos and the transition of maternal to zygotic control of embryo development are undertaking.
实验旨在评估兔输卵管上皮细胞(ROEC)或ROEC条件培养基促进体内或体外受精的大鼠卵子发育的能力。与ROEC共培养的体外受精卵子中有61.73%、体内受精卵子中有73.33%克服了2-细胞阻滞(图版I,表1和表2)。同样,在ROEC条件培养基中培养的体外受精卵子中有67.99%发育至2-细胞期以上,其中近一半发育至桑椹胚和囊胚期(表3)。通过使用35S-甲硫氨酸掺入和放射自显影方法,分别分子量为135 Kd、68 Kd、55 Kd、51 Kd和44 Kd的几种多肽从兔输卵管上皮细胞分泌出来并在ROEC条件培养基中被发现(图1)。通过确定是否有任何分泌蛋白与透明带结合来测试ROEC蛋白进入发育中胚胎的可能性。使用含125I的酰化剂标记法进行碘化的结果表明,68 Kd和55 Kd蛋白与体外与ROEC共培养24小时的大鼠卵子的透明带结合(图2)。关于这两种分泌蛋白是否是促进早期胚胎发育以及胚胎发育从母体控制向合子控制转变的关键因素的研究正在进行中。