Thakur Mahendra K, Kumar R C
Biochemistry & Molecular Biology Laboratory, Department of Zoology, Banaras Hindu University, Varanasi 221005, India.
Biogerontology. 2007 Oct;8(5):575-82. doi: 10.1007/s10522-007-9102-x. Epub 2007 Jul 6.
Androgen influences the function of central and peripheral nervous system and plays a crucial role in maintaining reproductive behaviors and neuroendocrine regulation. Such action is mediated by interaction of androgen receptor (AR) promoter with nuclear proteins, which are involved in transcriptional regulation of androgen responsive genes. We have analyzed the binding of AR core promoter to nuclear proteins from the cerebral cortex of adult and old mice of both sexes by electrophoretic mobility shift assay (EMSA) and characterized the bound protein by Southwestern blotting. EMSA showed that the binding of nuclear proteins declined in the cerebral cortex of intact old mice as compared to adult. Following gonadectomy, the binding was reduced in old male and adult female but increased in old female. In contrast, estradiol supplementation increased the binding in old male and adult female but decreased in old female. Southwestern blotting analysis revealed that a 40 kDa nuclear protein bound to the promoter and the binding pattern was similar to that observed in EMSA. Further characterization of this protein may help to explore the intricate mechanism that underlies the transcriptional regulation of androgen responsive genes during aging.
雄激素影响中枢和外周神经系统的功能,在维持生殖行为和神经内分泌调节中起关键作用。这种作用是由雄激素受体(AR)启动子与核蛋白相互作用介导的,这些核蛋白参与雄激素反应基因的转录调控。我们通过电泳迁移率变动分析(EMSA)分析了成年和老年雌雄小鼠大脑皮质中AR核心启动子与核蛋白的结合,并通过蛋白质印迹法对结合蛋白进行了表征。EMSA显示,与成年小鼠相比,完整老年小鼠大脑皮质中核蛋白的结合减少。去势后,老年雄性和成年雌性小鼠的结合减少,但老年雌性小鼠的结合增加。相反,补充雌二醇可增加老年雄性和成年雌性小鼠的结合,但老年雌性小鼠的结合减少。蛋白质印迹分析显示,一种40 kDa的核蛋白与启动子结合,其结合模式与EMSA中观察到的相似。对该蛋白的进一步表征可能有助于探索衰老过程中雄激素反应基因转录调控的复杂机制。