Bilodeau-Goeseels S, Schultz G A
Department of Medical Biochemistry, The University of Calgary, Alberta, Canada.
Biol Reprod. 1997 May;56(5):1323-9. doi: 10.1095/biolreprod56.5.1323.
The abundance of 28S, 18S, and 5S rRNA was measured by Northern blot techniques applied to RNA samples extracted from bovine oocytes and preattachment embryos produced by in vitro procedures. Total RNA content was estimated by comparing the intensity of hybridization signals of 28S and 18S rRNA probes to embryo RNA samples and to standard curves generated from bovine ovary or bovine oviduct cell RNA. RNA content declined from the oocyte to the morula stage (2.4 +/- 0.3 ng/oocyte, 1.7 +/- 0.5 ng/1-cell embryo, 2.2 +/- 0.9 ng/2- to 4-cell embryo, 0.8 +/- 0.2 ng/6- to 8-cell embryo, and 0.7 +/- 0.2 ng/morula). A marked increase in RNA content, based on levels of hybridization to 28S and 18S rRNA, was observed in blastocysts, in which values averaged 5.3 +/- 0.6 ng/embryo. On a relative basis, 5S rRNA abundance followed a pattern similar to that of 28S and 18S rRNA across the early development period to the blastocyst stage.
通过Northern印迹技术测定了28S、18S和5S rRNA的丰度，该技术应用于从牛卵母细胞和体外程序产生的着床前胚胎中提取的RNA样本。通过比较28S和18S rRNA探针与胚胎RNA样本以及从牛卵巢或牛输卵管细胞RNA生成的标准曲线的杂交信号强度，估计了总RNA含量。RNA含量从卵母细胞到桑葚胚阶段逐渐下降（2.4±0.3 ng/卵母细胞、1.7±0.5 ng/1细胞胚胎、2.2±0.9 ng/2至4细胞胚胎、0.8±0.2 ng/6至8细胞胚胎、0.7±0.2 ng/桑葚胚）。基于与28S和18S rRNA的杂交水平，在囊胚中观察到RNA含量显著增加，其中平均每个胚胎为5.3±0.6 ng。相对而言，在早期发育至囊胚阶段，5S rRNA丰度呈现出与28S和18S rRNA相似的模式。
