Changes in ribosomal ribonucleic acid content within in vitro-produced bovine embryos
Changes in ribosomal ribonucleic acid content within in vitro-produced bovine embryos
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DOI：
10.1095/biolreprod56.5.1323
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发表时间：
1997-05-01
影响因子：
3.6
通讯作者：
Schultz, GA
中科院分区：
文献类型：
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作者：
BilodeauGoeseels, S;Schultz, GA
通过北方印迹技术，应用于从牛卵母细胞和体外程序产生的附着前胚胎提取的RNA样品，测定28 S，18 S和5S rRNA的丰度。通过比较28 S和18 S rRNA探针与胚胎RNA样品的杂交信号强度以及与从牛卵巢或牛输卵管细胞RNA产生的标准曲线的杂交信号强度来估计总RNA含量。RNA含量从卵母细胞到桑椹胚阶段下降（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- 4-细胞胚胎，0.8 +/- 0.2 ng/6- 8-细胞胚胎和0.7 +/- 0.2 ng/桑椹胚）。根据与28 S和18 S rRNA的杂交水平，在囊胚中观察到RNA含量显着增加，平均值为5.3 +/- 0.6 ng/胚胎。在相对的基础上，5S rRNA丰度遵循的模式类似的28 S和18 S rRNA在早期发育阶段的囊胚阶段。
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.