Differential oocyte-specific expression of Cre recombinase activity in GDF-9-iCre, Zp3cre, and Msx2Cre transgenic mice
Differential oocyte-specific expression of Cre recombinase activity in GDF-9-iCre, Zp3cre, and Msx2Cre transgenic mice
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DOI：
10.1095/biolreprod.104.031757
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发表时间：
2004-11-01
影响因子：
3.6
通讯作者：
Cooney, AJ
中科院分区：
文献类型：
--
作者：
Lan, ZJ;Xu, XP;Cooney, AJ
利用Cre/loxP技术对卵巢基因进行卵母细胞特异性缺失，为了解它们在卵泡发生、卵发生和着床前胚胎发育过程中的生理作用提供了一个很好的工具。我们已经产生了一种转基因小鼠系，表达由小鼠生长分化因子-9 （GDF-9）启动子驱动的改良的Cre重组酶（iCre）。由此产生的转基因小鼠系被命名为GDF-9-iCre小鼠。在ROSA报告小鼠中，我们发现Cre重组酶在出生后卵巢中表达，但在心脏、肝脏、脾脏、肾脏和大脑中不表达。在卵巢内，Cre重组酶仅在原始卵泡和后期卵泡的卵母细胞中表达。GDF-9-iCre小鼠的iCre表达比Zp3Cre和Msx2Cre小鼠的Cre表达早，其中Cre基因分别由透明带蛋白3 （Zp3）启动子和同源盒基因Msx2启动子驱动。将携带GDF-9-iCre转基因基因的野生型雄性与杂合型GCNF雌性交配，未产生具有GCNF floxed等位基因的后代，这表明GDF-9-iCre小鼠可以在雌性种系中完全删除floxed GCNF等位基因。这些结果表明，GDF-9-iCre小鼠系为了解参与卵泡发生、卵发生和早期胚胎发育的卵母细胞表达基因的功能提供了一个很好的遗传工具。比较GDF-9-iCre、Zp3Cre和Msx2Cre转基因小鼠的Cre活性的发生，发现这三种转基因小鼠具有顺序的Cre活性，可以在卵泡发生的不同时间点使靶基因失活。
Oocyte-specific deletion of ovarian genes using Cre/loxP technology provides an excellent tool to understand their physiological roles during folliculogenesis, oogenesis, and preimplantation embryonic development. We have generated a transgenic mouse line expressing improved Cre recombinase (iCre) driven by the mouse growth differentiation factor-9 (GDF-9) promoter. The resulting transgenic mouse line was named GDF-9-iCre mice. Using the floxed ROSA reporter mice, we found that Cre recombinase was expressed in postnatal ovaries, but not in heart, liver, spleen, kidney, and brain. Within the ovary, the Cre recombinase was exclusively expressed in the oocytes of primordial follicles and follicles at later developmental stages. The expression of iCre of GDF-9-iCre mice was shown to be earlier than the Cre expression of Zp3Cre and Msx2Cre mice, in which the Cre gene is driven by zona pellucida protein 3 (Zp3) promoter and a homeobox gene Msx2 promoter, respectively, in the postnatal ovary. Breeding wild-type males with heterozygous floxed germ cell nuclear factor (GCNF) females carrying the GDF-9-iCre transgene did not produce any progeny having the floxed GCNF allele, indicating that complete deletion of the floxed GCNF allele can be achieved in the female germline by GDF-9-iCre mice. These results suggest that GDF-9-iCre mouse line provides an excellent genetic tool for understanding functions of oocyte-expressing genes involved in folliculogenesis, oogenesis, and early embryonic development. Comparison of the ontogeny of the Cre activities of GDF-9-iCre, Zp3Cre, and Msx2Cre transgenic mice shows there is sequential Cre activity of the three transgenes that will allow inactivation of a target gene at different points in folliculogenesis.