期刊
NEUROSCIENCE LETTERS
卷 593, 期 -, 页码 45-50出版社
ELSEVIER IRELAND LTD
DOI: 10.1016/j.neulet.2015.03.022
关键词
Adeno-associated virus; Glial fibrillary acidic protein; Promoter; Astrocyte
资金
- Natural Science Foundation of China [81271432]
- Natural Science Foundation of the Province of Gansu [145RJZA039]
We sought to demonstrate the in vivo transduction efficiency and tropism range in astrocytes of a combined-serotype adeno associated virus (AAV(2/5/7/8/9)). To control expression of enhanced green fluorescent protein (EGFP), a 1740bp glial fibrillary acidic protein (GFAP) promoter was obtained and ligated into vectors of each AAV serotype (2/5/7/8/9). Purified AAVs were then injected into the somatosensory cortex of C57BL/6J mice. Cell-type specific antibodies and subsequent immunofluorescence were used to identify astrocytes (GFAP), neurons (neuronal nuclear antigen, NeuN), microglia (ionized calcium-binding adapter molecule 1, Iba1), and oligodendrocytes (myelin basic protein, MBP), whereby, EGFP expression was measured in each cell type at 1-4 weeks post-injection. Our results indicated that the majority of astrocytes expressed EGFP, while only a small number of neurons expressed EGFP. Both microglia and oligodendrocytes lacked EGFP expression after viral injection. Quantitative analyses revealed that the percentage of EGFP-positive astrocytes was about 98% after viral injection, while the EGFP-positive neuronal percentage was less than 2%. Thus, this study shows that using a combined-serotype AAV carrying a 1740bp GFAP promoter results in successful, cell-type specific infection of the central nervous system, with robust gene expression in murine astrocytes. (C) 2015 Elsevier Ireland Ltd. All rights reserved.
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