Transgenic expression of enhanced green fluorescent protein enables direct visualization for physiological studies of vasopressin neurons and isolated nerve terminals of the rat
Transgenic expression of enhanced green fluorescent protein enables direct visualization for physiological studies of vasopressin neurons and isolated nerve terminals of the rat
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DOI：
10.1210/en.2004-0830
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发表时间：
2005-01-01
期刊：
影响因子：
4.8
通讯作者：
Murphy, D
中科院分区：
文献类型：
--
作者：
Ueta, Y;Fujihara, H;Murphy, D
我们已经产生了转基因大鼠表达精氨酸加压素（AVP）增强的绿色荧光蛋白（eGFP）融合基因。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eGFP基因的表达和强荧光。下丘脑-神经垂体束、分离的SON神经元和神经垂体中分离的轴突终末也显示出强烈的eGFP荧光。水分剥夺2d可增加SON和PVN中eGFP的荧光强度，但SCN中eGFP的荧光强度无明显变化。然后使用全细胞膜片钳技术记录体外脑切片制备物中特异性鉴定表达eGFP的SON、PVN和SCN AVP神经元的电活动。AVP-eGFP转基因大鼠是研究AVP分泌神经元在中枢神经系统中的生理作用以及AVP在活神经元及其轴突终末中的分泌调节动力学的独特的新工具。
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.