Birth Defects Center, University of Louisville Health Sciences Center, Louisville, KY 40202, USA.
Mol Reprod Dev. 2010 Jun;77(6):511-20. doi: 10.1002/mrd.21171.
Using an in silico approach, we identified a putative zinc finger domain-containing transcription factor (zinc finger protein 105, ZFP105) enriched in the adult mouse testis. RT-PCR analyses showed that Zfp105 was indeed highly expressed in adult mouse testis and that its expression was regulated during postnatal development. To further characterize Zfp105 expression, we generated a Zfp105:beta-galactosidase (LacZ) knock-in reporter mouse line (Zfp105(LacZ/+)) in which a Zfp105:LacZ fusion gene was expressed. Whole-mount LacZ analyses of adult Zfp105(LacZ/+) tissues showed robust LacZ staining in the testis, very weak staining in the ovary, and no staining in the spleen, liver, kidney, heart, lung, thymus, adrenal gland, uterus, or oviduct. Sectional LacZ staining showed that ZFP105 was highly expressed in pachytene spermatocytes. ZNF35, the human ortholog of Zfp105, was also highly expressed in human testis. Immunofluorescence analysis showed that ZNF35 was located primarily in the cytoplasm of male germ cells. More importantly, reduced male fertility was observed in adult Zfp105(LacZ/LacZ) mice. Histological studies showed the presence of undifferentiated spermatogenic cells in the lumen of seminiferous tubules at stage VII and in the epididymal lumen of adult Zfp105(LacZ/LacZ) mice. Taken together, our results suggest that ZFP105 is a male germ-cell factor and plays a role in male reproduction.
我们采用计算机模拟的方法,在成年小鼠睾丸中鉴定到一个富含锌指结构域的假定转录因子(锌指蛋白 105,ZFP105)。RT-PCR 分析表明,Zfp105 确实在成年小鼠睾丸中高度表达,其表达在出生后发育过程中受到调控。为了进一步研究 Zfp105 的表达特征,我们构建了一个 Zfp105:β-半乳糖苷酶(LacZ)的基因敲入报告鼠系(Zfp105(LacZ/+)),在该鼠系中,Zfp105:LacZ 融合基因得以表达。成年 Zfp105(LacZ/+)组织的整体 LacZ 分析显示,LacZ 染色在睾丸中强烈表达,在卵巢中极弱表达,在脾、肝、肾、心、肺、胸腺、肾上腺、子宫或输卵管中均无表达。组织切片 LacZ 染色显示,ZFP105 在粗线期精母细胞中高度表达。ZNF35 是 Zfp105 的人同源物,也在人睾丸中高度表达。免疫荧光分析显示,ZNF35 主要位于雄性生殖细胞的细胞质中。更为重要的是,成年 Zfp105(LacZ/LacZ)小鼠的雄性生育力明显降低。组织学研究显示,在成年 Zfp105(LacZ/LacZ)小鼠的生精小管管腔和附睾管腔中存在未分化的精原细胞。综上所述,我们的研究结果表明,ZFP105 是一种雄性生殖细胞因子,在雄性生殖中发挥作用。