Memili E, Dominko T, First N L
Endocrinology and Reproductive Physiology Program, Department of Animal Sciences, University of Wisconsin-Madison, 53706, USA.
Mol Reprod Dev. 1998 Sep;51(1):36-41. doi: 10.1002/(SICI)1098-2795(199809)51:1<36::AID-MRD4>3.0.CO;2-X.
The transition from the maternal to embryonic control of early embryonic development (MET) in mammals is not fully understood. The objective of this study was to determine the amount of transcriptional activity in immature oocytes containing germinal vesicle (GV), mature metaphase II arrested oocytes (MII), 2-, 4- and 8-cell bovine embryos by labeling with 35S-UTP followed by isolation of total RNA and autoradiography. Expression of counts per minute (CPM) per cell showed that incorporation of 35S-UTP in GV oocytes was significantly higher than the background (P < 0.01) and decreased sharply by the time the oocytes reached MII arrest. Incorporation significantly increased during the 2-cell stage and remained at the same level during the 4- and 8-cell stages. Uptake remained constant throughout different development stages (P > 0.05) with the highest variability observed during the 2-cell stage. When CPM were expressed per oocyte or embryo incorporation remained high at the GV stage, decreased to the background levels at the time of MII and increased again at the 2-cell stage. It remained at the same level during the 4-cell stage but increased significantly for the second time during the 8-cell stage. Uptake remained at the same level until the 8-cell stage when a significant increase was observed. The negative controls showed a significantly lower amount of incorporation compared to the positive control (P < 0.05). Similar results were observed by autoradiography. Our observations suggest that MET starts as early as the 2-cell stage in bovine embryos.
哺乳动物早期胚胎发育从母体控制向胚胎控制的转变(MET)尚未完全明晰。本研究的目的是通过用35S-UTP标记,随后分离总RNA并进行放射自显影,来确定含有生发泡(GV)的未成熟卵母细胞、成熟中期II期阻滞卵母细胞(MII)、2-、4-和8细胞期牛胚胎中的转录活性量。每个细胞的每分钟计数(CPM)表达显示,35S-UTP在GV卵母细胞中的掺入显著高于背景(P<0.01),并且在卵母细胞达到MII阻滞时急剧下降。掺入在2细胞期显著增加,并在4细胞期和8细胞期保持在相同水平。在不同发育阶段摄取保持恒定(P>0.05),在2细胞期观察到最高变异性。当以每个卵母细胞或胚胎表示CPM时,在GV期掺入仍然很高,在MII期降至背景水平,并在2细胞期再次增加。在4细胞期保持在相同水平,但在8细胞期第二次显著增加。摄取在8细胞期之前保持在相同水平,此时观察到显著增加。阴性对照显示掺入量明显低于阳性对照(P<0.05)。放射自显影观察到类似结果。我们的观察结果表明,MET在牛胚胎中最早在2细胞期开始。