Wu J T
J Reprod Fertil. 1980 Mar;58(2):455-61. doi: 10.1530/jrf.0.0580455.
Mouse embryos collected on Days 2 (2-cell) to 4 (early blastocyst) were treated with pronase to digest the zona pellucida and cultured at 37 degrees C for 3 h. These embryos and untreated Day-5 late blastocysts were exposed to [3H]Con A at 25, 50, 100 or 200 microgram/ml, 4 degrees C for 30 min, washed and counted for radioactivity. Other groups of embryos were incubated with [3H]Con A in the presence of 0.06 M-alpha-methyl-D-mannoside for the detection of non-specific binding. Scatchard plot analysis showed that the total number of Con A molecules bound/embryo increased with development, from 12 X 10(7) in 2-cell embryos to 19 X 10(7) in early blastocysts and to 150 X 10(7) in late blastocysts. The association constants decreased from 3 X 10(6) M-1 in 2-cell embryos to 0.83 X 10(6) M-1 in late blastocysts. The results suggest that the Con A binding sites (probably glycoproteins) on the embryo surface change quantitatively and qualitatively during preimplantation development.
收集第2天(2细胞期)至第4天(早期囊胚)的小鼠胚胎,用链霉蛋白酶处理以消化透明带,并在37℃培养3小时。将这些胚胎以及未经处理的第5天晚期囊胚暴露于浓度为25、50、100或200微克/毫升的[3H]刀豆球蛋白A中,在4℃下处理30分钟,然后洗涤并计数放射性。其他组胚胎在0.06Mα-甲基-D-甘露糖苷存在的情况下与[3H]刀豆球蛋白A一起孵育,以检测非特异性结合。Scatchard作图分析表明,结合到每个胚胎上的刀豆球蛋白A分子总数随着发育而增加,从2细胞期胚胎的12×10⁷增加到早期囊胚的19×10⁷,再到晚期囊胚的150×10⁷。亲和常数从2细胞期胚胎的3×10⁶M⁻¹降低到晚期囊胚的0.83×10⁶M⁻¹。结果表明,胚胎表面的刀豆球蛋白A结合位点(可能是糖蛋白)在植入前发育过程中发生了数量和质量上的变化。