Observation of the motion of a single molecule of neuronal nitric oxide synthase in the nucleus tractus solitarius

为了研究神经元型一氧化氮合酶(NNOS)单分子在孤束核(NTS)中的运动，我们尝试了将基因(绿色荧光蛋白和nNOS)导入孤束核。利用质粒载体在体内将基因导入脑内的简单方法尚未开发出来。方法：1.将含阳离子脂类的绿色荧光蛋白(GFP)融合Topo克隆载体(LipofextAMINE 2000)显微注射至NTS和舌下核，分别于注射后3、5、7、10天取脑，观察其荧光强度。5只大鼠脑内显微注射5天后，可观察到4个脑区的荧光。注射后第5天荧光强度最强。认为基因在转导后5天表达最强。接下来，我们将阳离子脂类微量注射到Lacz Vectors的大脑中。然而，我们不能观察到β-半乳糖。可以认为，使用小而大的载体进行脑内基因转导的效率可能较低。几种新的转基因药物已经迅速开发出来。我们尝试了一些新的转基因药物。我们可以使用某种新的转染剂来观察β-半乳糖。

In order to investigate the motion of a single molecule of neuronal nitric oxide synthase (nNOS) in the nucleus tractus solitarius (NTS), we tried gene (green fluorescent protein and nNOS) transduction into the NTS.1. The simple methods for gene transduction in vivo into the brain using plasmid vector have not been developed. We microinjected green fluorescent protein (GFP) Fusion TOPO Cloning Vectors with cationic lipids (LipofextAMINE 2000) into the NTS and the hypoglossal nucleus, and removed the brain three, five, seven and ten days after the microinjection, and observed the fluorescence.2. We could observe the fluorescence four brains microinjected out of five rats five days after the injection of plasmid vectors. The intensity of the fluorescence was the strongest five days after the injection. It is considered that the gene expression was the strongest five days after the transduction.3. Next, we microinjected LacZ Vectors with cationic lipids into the brain. However, we could not observe beta-gal. It may be considered that the efficiency of gene transduction into the brain using a little large vector may be low.4. Several new transfection drugs have been developed rapidly. We tried some new transfection drugs. We could observe the beta-gal using a certain new transfection drug.