Kubisch H Michael, Rasmussen Terri A, Johnson Kelly M
Department of Veterinary Medicine, Tulane National Primate Research Center, Covington, Louisiana 70435, USA.
Mol Reprod Dev. 2003 Jan;64(1):79-85. doi: 10.1002/mrd.10211.
A series of experiments were conducted to examine the pattern of production and secretion of interferon-tau (IFN-tau) by blastocysts following parthenogenetic activation of bovine oocytes. In the first experiment, 36.8, 24.1, and 33.2% of IVF-derived and parthenogenetically activated oocytes cultured in the presence or absence of a monolayer of buffalo rat liver cells, respectively, reached the blastocyst stage. Following individual culture of blastocysts, IFN-tau concentration in medium droplets was similar among the three groups, although IVF-derived blastocysts contained significantly more cells. In the second experiment, 156 IVF-derived blastocysts were sexed by PCR with 75 and 81, respectively, being male and female. IFN-tau secretion of these was compared to that of 70 parthenogenetic blastocysts. Female and parthenogenetic blastocysts produced significantly more IFN-tau than their male counterparts. In the third experiment, the ability of hatched blastocysts to form outgrowths and the pattern of their IFN-tau secretion were examined. Of the 48 IVF-derived blastocysts, 44 formed outgrowths compared to 41 of the 42 hatched parthenotes. Parthenogenetic outgrowths were significantly larger after 7 days, but this difference had disappeared after 14 days. IFN-tau secretion did not differ between the two groups. Lastly, sequence analyses of expressed mRNA from individual parthenogenetic blastocyst outgrowths showed four different transcript types which, based on their predicted amino acid sequence, belong to two subgroups, IFN-tau1 and IFN-tau3. In addition, one new transcript sequence was identified, encoding a new protein isoform.
进行了一系列实验，以研究牛卵母细胞孤雌激活后囊胚产生和分泌干扰素-τ（IFN-τ）的模式。在第一个实验中，分别在有或没有单层水牛大鼠肝细胞存在的情况下培养的体外受精（IVF）衍生和孤雌激活的卵母细胞，分别有36.8%、24.1%和33.2%发育到囊胚阶段。在对囊胚进行单独培养后，尽管IVF衍生的囊胚含有明显更多的细胞，但三组培养液滴中的IFN-τ浓度相似。在第二个实验中，通过聚合酶链反应（PCR）对156个IVF衍生的囊胚进行性别鉴定，其中75个为雄性，81个为雌性。将这些囊胚的IFN-τ分泌与70个孤雌囊胚的进行比较。雌性和孤雌囊胚产生的IFN-τ明显多于雄性囊胚。在第三个实验中，检查了孵化囊胚形成外植体的能力及其IFN-τ分泌模式。在48个IVF衍生的囊胚中，44个形成了外植体，而在42个孵化的孤雌胚中，有41个形成了外植体。孤雌外植体在7天后明显更大，但这种差异在14天后消失。两组之间的IFN-τ分泌没有差异。最后，对单个孤雌囊胚外植体表达的mRNA进行序列分析，显示出四种不同的转录本类型，根据其预测的氨基酸序列，它们属于两个亚组，即IFN-τ1和IFN-τ3。此外，还鉴定出一个新的转录本序列，编码一种新的蛋白质异构体。
