Bachvarova R, Cohen E M, De Leon V, Tokunaga K, Sakiyama S, Paynton B V
Department of Cell Biology and Anatomy, Cornell University Medical College, New York, NY 10021.
Development. 1989 Jul;106(3):561-5. doi: 10.1242/dev.106.3.561.
In order to measure the content of beta- and gamma-actin mRNA in mouse oocytes and ovulated eggs, Northern and slot blots were hybridized to complementary RNA probes transcribed from mouse isotype-specific cDNA sequences. The blots included samples of isotype-specific sense strand RNA standards prepared from the same cDNA sequences. Total actin mRNA content was estimated to be 40 fg per preovulatory full-grown oocyte or egg, consisting of one-third beta-actin mRNA and two-thirds gamma-actin mRNA. Ninety per cent of the actin mRNA is on polysomes in full-grown oocytes. The per cent of actin mRNA in polysomal mRNA is similar to the per cent of actin in newly synthesized proteins. Measurements on other developmental stages showed that, in mid-growth-phase oocytes, each actin mRNA reaches a level twofold higher than in full-grown oocytes. Thereafter, all modulations of the two isotypic mRNAs occur in parallel; that is, they are maintained at constant levels during the late growth phase (oocytes from females 8-14 days old); gradually degraded in oocytes that have completed their rapid growth phase (oocytes from females 15-18 days old), in maturing oocytes, and in 1- and 2-cell embryos; and deadenylated after about 7 h of progression into meiotic maturation.
为了测定小鼠卵母细胞和排卵卵中β-肌动蛋白和γ-肌动蛋白mRNA的含量,将Northern印迹和狭缝印迹与从小鼠同种型特异性cDNA序列转录的互补RNA探针杂交。印迹中包括从相同cDNA序列制备的同种型特异性有义链RNA标准品样本。据估计,每个排卵前的完全成熟卵母细胞或卵子中的总肌动蛋白mRNA含量为40 fg,其中三分之一为β-肌动蛋白mRNA,三分之二为γ-肌动蛋白mRNA。90%的肌动蛋白mRNA存在于完全成熟卵母细胞的多核糖体上。多核糖体mRNA中肌动蛋白mRNA的百分比与新合成蛋白质中肌动蛋白的百分比相似。对其他发育阶段的测量表明,在生长中期的卵母细胞中,每个肌动蛋白mRNA的水平比完全成熟卵母细胞中的高两倍。此后,两种同种型mRNA的所有调节都是平行发生的;也就是说,它们在生长后期(8-14日龄雌性小鼠的卵母细胞)保持恒定水平;在完成快速生长阶段的卵母细胞(15-18日龄雌性小鼠的卵母细胞)、成熟卵母细胞以及1-细胞和2-细胞胚胎中逐渐降解;并在进入减数分裂成熟约7小时后去腺苷酸化。