Single-stranded adeno-associated virus serotype 9 mediated gene expression in astrocyte via intravenous delivery
10.3969/j.issn.1674-8115.2020.08.006
- VernacularTitle: 经静脉注射的单链9型腺相关病毒介导基因在星形胶质细胞表达
- Author:
Xiao-Dan WANG
1
Author Information
1. Ruijin Hospital North, Shanghai Jiao Tong University School of Medicine
- Publication Type:Journal Article
- Keywords:
Adeno-associated virus serotype 9 (AAV9);
Astrocyte;
Cerebral ischemia;
Hypoxia-responsive element (HRE)
- From:
Journal of Shanghai Jiaotong University(Medical Science)
2020;40(8):1036-1040
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To investigate the expression of LacZ gene mediated by intravenous injection of the single-stranded adeno-associated virus serotype 9 (ssAAV9) containing hypoxia-responsive element (HRE) promoter in the cerebral ischemic area, and further identify the types of brain cells that can be transfected by the vector. Methods: A mouse model of permanent left distal middle cerebral artery occlusion (dMCAO) was established. The expression of hypoxia-inducible factor-1 (HIF-1) in cerebral ischemic area was detected at 1 and 5 days after ischemia. The ssAAV vector containing HRE-regulated LacZ gene was packaged into the capsid of AAV9 virus (AAV9-H9LacZ), and AAV9-H9LacZ was injected into mice through jugular vein 1 h after the establishment of dMCAO model. Five days after injection of AAV9-H9LacZ, X-gal staining was used to detect the expression of β-galactosidase (β-gal) encoded by the LacZ gene in the ischemic area and liver tissue. Immunofluorescence double staining was used to detect the expression of β-gal in astrocyte, neurons and vascular endothelial cells. Results: The expression of HIF-1 was increased 1 and 5 days after ischemia. β-gal was mainly expressed in ischemic penumbra of mice injected with AAV9-H9LacZ. There was no positive expression of β-gal in the liver tissue of mice. β-gal was mainly co-expressed with glial fibrillary acidic protein as an astrocyte-specific marker, and a little of β-gal was co-expressed with neuron-specific nuclear protein. Conclusion: After cerebral ischemia, intravenous injection of AAV9-H9LacZ can effectively mediate gene expression in astrocyte in the cerebral ischemia area. HRE can effectively control the expression of the LacZ gene in cerebral ischemia.
