In vivo transduction of murine cerebellar Purkinje cells by HIV-derived lentiviral vectors
In vivo transduction of murine cerebellar Purkinje cells by HIV-derived lentiviral vectors
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DOI：
10.1016/j.brainres.2006.01.104
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发表时间：
2006-04
期刊：
影响因子：
2.9
通讯作者：
T. Torashima;S. Okoyama;T. Nishizaki;H. Hirai
中科院分区：
文献类型：
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作者：
T. Torashima;S. Okoyama;T. Nishizaki;H. Hirai
小脑浦肯野细胞是运动学习和运动协调的关键元素，因此，阐明浦肯野细胞整合信息和控制小脑功能的机制非常重要。将基因转移到神经元，然后评估其对神经功能的影响，是检测基因功能的有效方法。然而，这种方法还没有被完全应用于小脑的研究，因为腺病毒载体是最常用的体内基因转移载体，对浦肯野细胞的亲和力很低。在这项研究中，我们使用了人类免疫缺陷病毒(HIV)衍生的慢病毒载体，并检测了该载体在小脑中的转导情况。携带GFP基因的慢病毒载体被注射到小脑皮质。注射后7天，Purkinje细胞被有效地转导，而对细胞活力和突触功能没有明显影响。GFP在其他皮质中间神经元和Bergmann胶质细胞中也有表达，尽管表达效率较低。与其他病毒载体的研究结果相反，在小脑皮质外没有观察到转导细胞。因此，当HIV来源的慢病毒载体被注射到小脑皮质时，转导仅限于小脑皮质中的细胞，对浦肯野细胞的趋向性最高。这些结果表明，HIV衍生的慢病毒载体对于研究浦肯野细胞的基因功能以及作为治疗影响浦肯野细胞的疾病的基因治疗工具是有用的。
Cerebellar Purkinje cells are key elements in motor learning and motor coordination, and therefore, it is important to clarify the mechanisms by which Purkinje cells integrate information and control cerebellar function. Gene transfer into neurons, followed by the assessment of the effects on neural function, is an effective approach for examining gene function. However, this method has not been used fully in the study of the cerebellum because adenovirus vectors, the vectors most commonly used for in vivo gene transfer, have very low affinity for Purkinje cells. In this study, we used a human immunodeficiency virus (HIV)-derived lentiviral vector and examined the transduction profile of the vector in the cerebellum. A lentiviral vector carrying the GFP gene was injected into the cerebellar cortex. Seven days after the injection, Purkinje cells were efficiently transduced without significant influence on the cell viability and synaptic functions. GFP was also expressed, though less efficiently, in other cortical interneurons and Bergmann glias. In contrast to reported findings with other viral vectors, no transduced cells were observed outside of the cerebellar cortex. Thus, when HIV-derived lentiviral vectors were injected into the cerebellar cortex, transduction was limited to the cells in the cerebellar cortex, with the highest tropism for Purkinje cells. These results suggest that HIV-derived lentiviral vectors are useful for the study of gene function in Purkinje cells as well as for application as a gene therapy tool for the treatment of diseases that affect Purkinje cells.