Lei Zhenmin, Lin Jing, Li Xian, Li Shengqiang, Zhou Huaxin, Araki Yoshihiko, Lan Zi-Jian
Department of OB/GYN and Women's Health, University of Louisville Health Sciences Center, Louisville, Kentucky, USA.
Genesis. 2010 Dec;48(12):717-22. doi: 10.1002/dvg.20675. Epub 2010 Oct 30.
We have generated a transgenic mouse line that expresses improved Cre recombinase (iCre) under the control of the testis-expressed gene 101 (Tex101) promoter. This transgenic mouse line was named Tex101-iCre. Using the floxed ROSA reporter mice, we found that robust Cre recombinase activity was detected in postnatal testes with weak or no activity in other tissues. Within the testis, Cre recombinase was active in spermatogenic cells as early as the prospermatogonia stage at day 1 after birth. In 30- and 60-day-old mice, positive Cre recombinase activity was detected not only in prospermatogonia but also in spermatogenic cells at later stages of spermatogenesis. There was little or no Cre activity in interstitial cells. Breeding wild-type females with homozygous floxed fibroblast growth factor receptor 2 (Fgfr2) males carrying the Tex101-iCre transgene did not produce any progeny with the floxed Fgfr2 allele. All the progeny inherited a recombined Fgfr2 allele, indicating that complete deletion of the floxed Fgfr2 allele by Tex101-iCre can be achieved in the male germline. Furthermore, FGFR2 protein was not detected in spermatocytes and spermatids of adult Fgfr2(fl/fl) ;Tex101-iCre mice. Taken together, our results suggest that the Tex101-iCre mouse line allows the inactivation of a floxed gene in spermatogenic cells in adult mice, which will facilitate the functional characterization of genes in normal spermatogenesis and male fertility.
我们构建了一个转基因小鼠品系,该品系在睾丸特异性表达基因101(Tex101)启动子的控制下表达改良型Cre重组酶(iCre)。这个转基因小鼠品系被命名为Tex101-iCre。利用携带loxP侧翼序列的ROSA报告基因小鼠,我们发现出生后的睾丸中可检测到强大的Cre重组酶活性,而在其他组织中活性较弱或没有活性。在睾丸内,Cre重组酶早在出生后第1天的精原细胞阶段就在生精细胞中具有活性。在30日龄和60日龄的小鼠中,不仅在精原细胞中检测到阳性Cre重组酶活性,在精子发生后期的生精细胞中也检测到了活性。间质细胞中几乎没有或没有Cre活性。将野生型雌性小鼠与携带Tex101-iCre转基因的纯合loxP侧翼序列成纤维细胞生长因子受体2(Fgfr2)雄性小鼠杂交,没有产生任何带有loxP侧翼序列Fgfr2等位基因的后代。所有后代都继承了重组的Fgfr2等位基因,这表明Tex101-iCre可以在雄性生殖系中实现对loxP侧翼序列Fgfr2等位基因的完全缺失。此外,在成年Fgfr2(fl/fl) ;Tex101-iCre小鼠的精母细胞和精子细胞中未检测到FGFR2蛋白。综上所述,我们的结果表明,Tex101-iCre小鼠品系能够使成年小鼠生精细胞中的loxP侧翼序列基因失活,这将有助于对正常精子发生和雄性生育力相关基因的功能进行表征。