Department of Physiology, School of Medicine, University of Occupational and Environmental Health, 1-1 Iseigaoka, Yahatanishi-ku, Kitakyushu 807-8555, Japan.
Endocrinology. 2011 Jul;152(7):2768-74. doi: 10.1210/en.2011-0006. Epub 2011 May 3.
We have generated rats bearing an oxytocin (OXT)-monomeric red fluorescent protein 1 (mRFP1) fusion transgene. The mRFP1 fluorescence was highly visible in ventral part of the supraoptic nucleus (SON) and the posterior pituitary in a whole mount. mRFP1 fluorescence in hypothalamic sections was also observed in the SON, the paraventricular nucleus (PVN), and the internal layer of the median eminence. Salt loading for 5 d caused a marked increase in mRFP1 fluorescence in the SON, the PVN, the median eminence, and the posterior pituitary. In situ hybridization histochemistry revealed that the expression of the mRNA encoding the OXT-mRFP1 fusion gene was observed in the SON and the PVN of euhydrated rats and increased dramatically after chronic salt loading. The expression of the endogenous OXT and the arginine vasopressin (AVP) genes were significantly increased in the SON and the PVN after chronic salt loading in both nontransgenic and transgenic rats. These responses were not different between male and female rats. Compared with nontransgenic rats, euhydrated and salt-loaded male and female transgenic rats showed no significant differences in plasma osmolality, sodium concentration, OXT, and AVP levels. Finally, we succeeded in generating a double-transgenic rat that expresses both the OXT-mRFP1 fusion gene and the AVP-enhanced green fluorescent protein fusion gene. Our new transgenic rats are valuable new tools to study the physiology of the hypothalamo-neurohypophysial system.
我们生成了携带催产素(OXT)单体红色荧光蛋白 1(mRFP1)融合转基因的大鼠。在整体标本中,mRFP1 荧光在视上核(SON)的腹侧部分和垂体后叶高度可见。在 SON、室旁核(PVN)和正中隆起的内层中也观察到下丘脑切片中的 mRFP1 荧光。5 天的盐负荷导致 SON、PVN、正中隆起和垂体后叶中的 mRFP1 荧光明显增加。原位杂交组织化学显示,编码 OXT-mRFP1 融合基因的 mRNA 在正常水合大鼠的 SON 和 PVN 中表达,并在慢性盐负荷后显著增加。在慢性盐负荷后,SON 和 PVN 中的内源性 OXT 和精氨酸加压素(AVP)基因的表达在非转基因和转基因大鼠中均显著增加。这些反应在雄性和雌性大鼠之间没有差异。与非转基因大鼠相比,正常水合和盐负荷的雄性和雌性转基因大鼠的血浆渗透压、钠浓度、OXT 和 AVP 水平没有显著差异。最后,我们成功生成了一种同时表达 OXT-mRFP1 融合基因和 AVP 增强型绿色荧光蛋白融合基因的双转基因大鼠。我们的新转基因大鼠是研究下丘脑神经垂体系统生理学的有价值的新工具。