Ueta Yoichi, Fujihara Hiroaki, Serino Ryota, Dayanithi Govindan, Ozawa Hitoshi, Matsuda Ken-ichi, Kawata Mitsuhiro, Yamada Junko, Ueno Shinya, Fukuda Atsuo, Murphy David
Department of Physiology, School of Medicine, University of Occupational and Environmental Health, Kitakyushu 807-8555, Japan.
Endocrinology. 2005 Jan;146(1):406-13. doi: 10.1210/en.2004-0830. Epub 2004 Sep 16.
We have generated transgenic rats expressing an arginine vasopressin (AVP)-enhanced green fluorescent protein (eGFP) fusion gene. The expression of the eGFP gene and strong fluorescence were observed in the supraoptic nucleus (SON), the paraventricular nucleus (PVN), and the suprachiasmatic nucleus (SCN) in transgenic rats. The hypothalamo-neurohypophyseal tract, isolated SON neurons, and isolated axon terminals in the neurohypophysis also showed robust eGFP fluorescence. Water deprivation for 2 d increased the fluorescence of the eGFP in the SON and the PVN but not the SCN. The whole-cell patch-clamp technique was then used to record the electrical activities specifically identifying eGFP-expressing SON, PVN, and SCN AVP neurons in in vitro brain slice preparations. The AVP-eGFP transgenic rats are a unique new tool with which to study the physiological role of AVP-secreting neurons in the central nervous system and the dynamics of the regulation of AVP secretion in the living neurons and their axon terminals.
我们已经培育出了表达精氨酸加压素(AVP)-增强型绿色荧光蛋白(eGFP)融合基因的转基因大鼠。在转基因大鼠的视上核(SON)、室旁核(PVN)和视交叉上核(SCN)中观察到了eGFP基因的表达及强烈荧光。下丘脑-神经垂体束、分离出的SON神经元以及神经垂体中分离出的轴突终末也显示出强烈的eGFP荧光。禁水2天增加了SON和PVN中eGFP的荧光,但SCN中未增加。然后使用全细胞膜片钳技术记录电活动,以特异性鉴定体外脑片制备物中表达eGFP的SON、PVN和SCN AVP神经元。AVP-eGFP转基因大鼠是一种独特的新工具,可用于研究中枢神经系统中分泌AVP的神经元的生理作用以及活体神经元及其轴突终末中AVP分泌调节的动力学。