Hu Yuanxin, Zhou Zongyao, Xu Chen, Shang Quan, Zhang You-Duan, Zhang Yong-Lian
State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
Endocrinology. 2003 Apr;144(4):1612-9. doi: 10.1210/en.2002-220915.
Germ cell nuclear factor (GCNF), a nuclear orphan receptor, involved in spermatogenesis, neurogenesis, differentiation, and embryo development, was highly expressed with two transcripts (7.4 and 2.3 kb) in mouse testis and with only one transcript (7.4 kb) slightly expressed in brain, liver, and kidney. The 2.3-kb transcript was restricted to round spermatids at stages VII and VIII of the spermatogenic cycle. The present report demonstrated its expression in epididymis as well, but at a very low level. Northern blot analysis showed two transcripts: a common 7.4-kb transcript and a unique 3.1-kb transcript. The expression levels of both GCNF transcripts in epididymis were down-regulated by androgen, as observed in castrated animals and aged mice. Polyclonal antisera against GCNF protein were raised. Western blot analysis showed the presence of only one band in total protein extracts from either mouse testis or epididymis. It indicated that the two mRNAs (7.4 and 3.1 kb) encode for the same protein as in testis. Fluorescent immunohistochemical staining and in situ hybridization showed that its expression was in the principal cell abundant in the corpus region. It implies that some androgen-regulated gene expressions located at the corpus principal cells might be controlled by GCNF.
生殖细胞核因子（GCNF）是一种核孤儿受体，参与精子发生、神经发生、分化和胚胎发育，在小鼠睾丸中以两种转录本（7.4和2.3 kb）高度表达，而在脑、肝和肾中仅有一种转录本（7.4 kb）轻微表达。2.3 kb的转录本局限于生精周期VII和VIII期的圆形精子细胞。本报告还证明了其在附睾中的表达，但水平非常低。Northern印迹分析显示有两种转录本：一种常见的7.4 kb转录本和一种独特的3.1 kb转录本。如在去势动物和老年小鼠中观察到的那样，附睾中两种GCNF转录本的表达水平均受雄激素下调。制备了针对GCNF蛋白的多克隆抗血清。Western印迹分析显示，从小鼠睾丸或附睾的总蛋白提取物中仅存在一条带。这表明这两种mRNA（7.4和3.1 kb）在睾丸中编码相同的蛋白质。荧光免疫组织化学染色和原位杂交显示，其表达位于体部区域丰富的主细胞中。这意味着位于体部主细胞中的一些雄激素调节基因表达可能受GCNF控制。
