title: Specific gene expression in mouse cortical astrocytes is mediated by a 1740bp-GFAP promoter-driven combined adeno-associated virus 2/5/7/8/9.
abstract: We sought to demonstrate the in vivo transduction efficiency and tropism range in astrocytes of a combined-serotype adeno associated virus (AAV2/5/7/8/9). 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.
authors.affiliation: Department of Neurology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China; Department of Neurology, Lanzhou General Hospital of PLA, Lanzhou 730050, PR China.
authors.forename: Xiandong
authors.initials: X
authors.lastname: Meng
authors.affiliation: Department of Neurology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Yang
authors.affiliation: Department of Neurology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China.
authors.forename: Tangpeng
authors.initials: T
authors.lastname: Ouyang
authors.affiliation: Shanghai SBO Medical Biotechnology Co. Ltd., Shanghai 201203, China.
authors.forename: Bing
authors.initials: B
authors.lastname: Liu
authors.affiliation: Department of Neurology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China.
authors.forename: Chen
authors.initials: C
authors.lastname: Wu
authors.affiliation: Department of Neurology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China. Electronic address: jiangwen@fmmu.edu.cn.
authors.forename: Wen
authors.initials: W
authors.lastname: Jiang
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000166:cytology / Q000378:metabolism*; D000229:Dependovirus / Q000235:genetics*; D015870:Gene Expression; D005904:Glial Fibrillary Acidic Protein / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D008297:Male; D008810:Mice, Inbred C57BL; D017628:Microglia / Q000166:cytology / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D009836:Oligodendroglia / Q000166:cytology / Q000378:metabolism; D011401:Promoter Regions, Genetic; D013003:Somatosensory Cortex / Q000166:cytology / Q000378:metabolism*; D014161:Transduction, Genetic
chemical_list: D005904:Glial Fibrillary Acidic Protein; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
keywords: Adeno-associated virus; Astrocyte; Glial fibrillary acidic protein; Promoter