PKC对小鼠受精卵发育的调控作用

The Regulation of PKC in Fertilized Mouse Eggs

为研究 TPA及 PKC的反义寡核苷酸对 1 -细胞期鼠受精卵发育的影响 ,采用免疫细胞化学法标记 PKC(α及 β亚型 ) ,并用激光扫描共聚焦显微镜测定卵内 PKC荧光强度 ;同时利用显微注射法注射 PKC的反义寡核苷酸 ,观察其对受精卵分裂的影响 . 1 0 0 μg/ L TPA对 1 -细胞期受精卵的发育具有完全抑制作用 .TPA处理 1 2 h后 ,对照组受精卵停留在 1 -细胞期 ,而未经 TPA处理的1 -细胞期卵可以分裂到 2 -细胞期 .共焦激光显示实验组与对照组相比 ,PKC(α、β亚型 )荧光强度均有下降 (P<0 .0 1 ) .显微注射 PKC antisenseα及 antisenseβ的受精卵 ,分别只有 1 4 .2 %和 3.33%的卵可以发育到 2 -细胞期 .与对照组 (注射 M2培养液 )差异显著 (P<0 .0 1 ) .结果表明 ,(1 ) TPA长期处理 1 -细胞期受精卵 ,抑制 1 -细胞期卵分裂到 2 -细胞期 ;(2 ) PKC的反义寡核苷酸 (α及β亚型 )可以抑制小鼠 1

The effects of TPA and antisense oligodeoxynucleotide of PKC on the development of 1 cell stage fertilized mouse eggs were explored.After stained by the methods of immunocytochemistry,the fluorescence intensity of PKC in eggs was measured by Laser scanning confocal microscope;the percentage of 1 cell stage eggs division was observed after microinjecting the antisense oligodeoxynucleotide of PKC.The division of 1 cell stage eggs could be inhibited thoroughly after the treatment of 100 μg/L TPA.The experimental group remained at 1 cell stage,while the control group could divide into 2 cell stage.The fluorescence intensities of two groups were significantly different ( P <0.01).Only 14.2% and 3.33% eggs could develop into 2 cell stage embryos after microinjecting antisense α and antisense β( P <0.01).The result suggests that 1 cell stage eggs can’t divide into 2 cell stage after long time treatment of TPA and antisense oligodeoxynucleotide of PKC(α、β) can inhibit the division of 1 cell stage eggs.