Ebert K M, Paynton B V, McKnight G S, Brinster R L
J Embryol Exp Morphol. 1984 Dec;84:91-103.
Growing mouse oocytes and fertilized ova were injected with chicken ovalbumin messenger RNA (mRNAov) and chicken conalbumin mRNA (mRNAcon) and cultured in vitro. Estimation of mRNAov and mRNAcon stability by hybridization of cDNAov and cDNAcon to extracted mRNA from injected oocytes and fertilized ova indicated a half-life of 147 and 366 h in the oocyte and 5 and 3 h in the fertilized ovum respectively. Stability of mRNAov was similar in the fertilized and unfertilized ovum. Oocytes injected with chicken ovalbumin mRNA were also labelled with [3H]leucine and ovalbumin synthesis was measured by immunoprecipitation. The amount of ovalbumin synthesized during the initial 7 h was less than during the period of 18-25 or 66-73 h postinjection. The greatest percentage of ovalbumin to total protein synthesis occurred between 66-73 h. Oocytes secreted 12% of the synthesized ovalbumin during each of the 7 h periods (0-7, 18-25 and 66-73 h) indicating a stable mechanism for secretion throughout the culture period. These studies demonstrate: a dramatic difference in stability of injected mRNA between the growing oocyte and the unfertilized or fertilized ovum, and a gradual increase in the translation of injected mRNA by the growing oocyte during in vitro culture.
将生长中的小鼠卵母细胞和受精卵注射鸡卵清蛋白信使核糖核酸（mRNAov）和鸡伴清蛋白信使核糖核酸（mRNAcon），并进行体外培养。通过将cDNAov和cDNAcon与从注射的卵母细胞和受精卵中提取的mRNA杂交来估计mRNAov和mRNAcon的稳定性，结果表明，在卵母细胞中其半衰期分别为147小时和366小时，在受精卵中分别为5小时和3小时。mRNAov在受精卵和未受精卵中的稳定性相似。用鸡卵清蛋白信使核糖核酸注射的卵母细胞也用[3H]亮氨酸进行标记，并通过免疫沉淀法测定卵清蛋白的合成。在注射后的最初7小时内合成的卵清蛋白量少于注射后18 - 25小时或66 - 73小时期间。卵清蛋白占总蛋白合成的最大百分比出现在66 - 73小时之间。卵母细胞在每个7小时时间段（0 - 7小时、18 - 25小时和66 - 73小时）内分泌了12%的合成卵清蛋白，表明在整个培养期间存在稳定的分泌机制。这些研究表明：生长中的卵母细胞与未受精卵或受精卵之间，注射的mRNA稳定性存在显著差异；在体外培养过程中，生长中的卵母细胞对注射的mRNA的翻译逐渐增加。
