Hipkaeo Wiphawi, Sakulsak Natthiya, Wakayama Tomohiko, Yamamoto Miyuki, Nakaya Masa-Aki, Keattikunpairoj Sunisa, Kurobo Miho, Iseki Shoichi
Department of Histology and Embryology, Graduate School of Medical Science, Kanazawa University.
Tohoku J Exp Med. 2008 Mar;214(3):231-45. doi: 10.1620/tjem.214.231.
In the submandibular gland (SMG) of mice, a duct portion called the granular convoluted tubule (GCT) is developed preferentially in males with puberty. This sexual dimorphism is androgen-dependent, but the underlying molecular mechanisms are unclear. We have demonstrated that the expression of a transcription factor JunD is regulated in association with the androgen-induced differentiation of GCT cells from striated duct (SD) cells. Menin, a nuclear protein encoded by the MEN1 tumor-suppressor gene, is known to bind JunD, thereby inhibiting its activity. In the present study, we examined the expression of menin in the mouse SMG by use of Northern blotting, Western blotting, and immunohistochemistry. Immunoreactivity for menin was higher in the female than male gland, and localized to the nuclei of intercalated duct cells and a subpopulation of SD cells. In contrast, GCT cells in males appeared negative for menin. The levels of menin in the SMG were increased with castration in males and decreased by repeated administration of testosterone to females or to castrated males. After a single administration of testosterone to females, many SD cells newly gained nuclear menin, which was lost as the cells converted to GCT cells by 48 hrs. These patterns of the expression and localization of menin were quite similar to those of JunD. Furthermore, the coimmunoprecipitation analysis of the SMG homogenates indicated that menin binds JunD in vivo. The present study suggests that the JunD-menin complex plays significant roles in the androgen-dependent differentiation of the duct system in the mouse SMG.
在小鼠的下颌下腺(SMG)中,一个称为颗粒曲管(GCT)的导管部分在青春期的雄性小鼠中优先发育。这种性别二态性依赖雄激素,但潜在的分子机制尚不清楚。我们已经证明,转录因子JunD的表达与雄激素诱导的GCT细胞从纹状管(SD)细胞分化相关。Menin是一种由MEN1肿瘤抑制基因编码的核蛋白,已知它能与JunD结合,从而抑制其活性。在本研究中,我们通过Northern印迹、Western印迹和免疫组织化学检测了menin在小鼠SMG中的表达。女性腺体中menin的免疫反应性高于男性腺体,且定位于闰管细胞和一部分SD细胞的细胞核中。相反,雄性的GCT细胞中menin呈阴性。雄性小鼠去势后,SMG中menin的水平升高,而对雌性或去势雄性小鼠反复给予睾酮后,menin水平降低。对雌性小鼠单次给予睾酮后,许多SD细胞新获得了核内menin,但在48小时后,随着这些细胞转化为GCT细胞,menin消失。menin的这些表达和定位模式与JunD的非常相似。此外,对SMG匀浆进行的免疫共沉淀分析表明,menin在体内与JunD结合。本研究表明,JunD - menin复合物在小鼠SMG导管系统的雄激素依赖性分化中起重要作用。