Lira S A, Kinloch R A, Mortillo S, Wassarman P M
Department of Cell and Developmental Biology, Roche Research Center, Nutley, NJ 07110.
Proc Natl Acad Sci U S A. 1990 Sep;87(18):7215-9. doi: 10.1073/pnas.87.18.7215.
The gene encoding the mouse egg primary receptor for sperm, a zona pellucida glycoprotein called ZP3, is expressed exclusively in growing oocytes within ovaries of sexually immature and mature female mice. We have constructed a transgene in which 6.5 kilobases of ZP3 gene 5'-flanking sequence is fused to the coding region of the firefly luciferase gene, and we have generated four independent lines of transgenic mice. In these animals, the transgene is expressed exclusively in ovaries. Furthermore, within ovaries, expression is confined to growing oocytes, and luciferase activity can be detected by assaying individual, isolated oocytes. The pattern of firefly luciferase expression during oocyte growth is similar to that observed in previous studies of ZP3 expression during oogenesis in mice. Observations reported here strongly suggest that cis-acting elements present in the ZP3 gene 5'-flanking region regulate oocyte-specific and, therefore, sex-specific expression of the sperm receptor gene during mouse development. They also suggest that such elements can be used to direct expression of cloned genes specifically to oocytes of transgenic mice and to evaluate the effects of such expression on various aspects of early mammalian development.
编码小鼠卵子精子初级受体的基因是一种名为ZP3的透明带糖蛋白,仅在未成熟和成熟雌性小鼠卵巢内的生长中的卵母细胞中表达。我们构建了一个转基因,其中6.5千碱基的ZP3基因5'侧翼序列与萤火虫荧光素酶基因的编码区融合,并产生了四个独立的转基因小鼠品系。在这些动物中,转基因仅在卵巢中表达。此外,在卵巢内,表达局限于生长中的卵母细胞,并且通过检测单个分离的卵母细胞可以检测到荧光素酶活性。卵母细胞生长过程中萤火虫荧光素酶的表达模式与之前小鼠卵子发生过程中ZP3表达的研究中观察到的模式相似。这里报道的观察结果强烈表明,ZP3基因5'侧翼区域中存在的顺式作用元件在小鼠发育过程中调节卵母细胞特异性以及因此的精子受体基因的性别特异性表达。它们还表明,此类元件可用于将克隆基因的表达特异性地导向转基因小鼠的卵母细胞,并评估这种表达对早期哺乳动物发育各个方面的影响。