Lentiviral transduction of murine oligodendrocytes in vivo
Lentiviral transduction of murine oligodendrocytes in vivo
复制标题
DOI：
10.1002/jnr.20626
复制
发表时间：
2005-11-01
影响因子：
4.2
通讯作者：
Goldberg, MP
中科院分区：
文献类型：
--
作者：
McIver, SR;Lee, CS;Goldberg, MP
慢病毒载体广泛用于指导体内有效的基因转移。我们研究了立体定位显微注射四种慢病毒结构后成年小鼠白色物质中的细胞特异性表达。我们合成了水泡性口炎病毒糖蛋白（VSV-G）假型慢病毒，其具有两个启动子（巨细胞病毒（CMV）或髓鞘碱性蛋白（MBP））和两个报告序列（胞质增强型绿色荧光蛋白（eGFP）或质膜靶向eGFP（人淋巴细胞特异性蛋白酪氨酸激酶[Lck]-eGFP））的组合。对于所有构建体，脑内注射到外侧胼胝体导致前脑白色物质神经胶质细胞中广泛的GFP表达。在注射后3天内观察到强烈的细胞GFP荧光，并持续至少28天。CMV启动子在多种神经胶质细胞类型中指导GFP表达，而MBP启动子特异性地将GFP靶向少突胶质细胞。膜靶向Lck-eGFP构建体的表达清楚地标记了少突胶质细胞的单个髓鞘形成过程。在MBP启动子下表达eGFP或Lck-eGFP的慢病毒构建体在完整的白色物质中提供了少突胶质细胞形态的极好的可视化，并且可以证明对于在体内向少突胶质细胞递送另外的感兴趣的基因是有价值的。(c)2005 Wiley-Liss，Inc.
Lentiviral vectors are used widely to direct efficient gene transfer in vivo. We examined cell-specific expression in adult murine white matter after stereotaxic microinjection of four lentiviral constructs. We synthesized vesicular stomatitis virus glycoprotein (VSV-G) pseudotyped lentiviruses with combinations of two promoters, cytomegalovirus (CMV) or myelin basic protein (MBP), and two reporter sequences, cytosolic enhanced green fluorescent protein (eGFP) or a plasma membrane-targeted eGFP (human lymphocyte-specific protein tyrosine kinase [Lck]-eGFP). For all constructs, intracerebral injections to lateral corpus callosum resulted in widespread GFP expression in forebrain white matter glial cells. Intense cellular GFP fluorescence was observed within 3 days after injection and lasted for at least 28 days. The CMV promoter directed GFP expression in multiple glial cell types, whereas the MBP promoter targeted GFP specifically to oligodendrocytes. Expression of the membrane-targeted Lck-eGFP construct distinctly labeled individual myelinating processes of oligodendrocytes. Lentiviral constructs expressing eGFP or Lck-eGFP under the MBP promoter provide excellent visualization of oligodendrocyte morphology in intact white matter, and may prove valuable for delivering additional genes of interest to oligodendrocytes in vivo. (c) 2005 Wiley-Liss, Inc.