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Updated: Sep 20, 2026
Direct Intrathecal Injection of Recombinant Adeno-associated Viruses in Adult Mice
Published on: February 15, 2019
[Recombinant adenovirus carrying glial cell line-derived neurotrophic factor gene protect midbrain dopaminergic
Hong Gao1, Jianjun Wang, Wei Zhang
1Neuroscience Research Institute, Peking University, Department of Neurobiology, Peking University School of Basic Medical Sciences, Beijing 100083, China.
Objective:
To observe the protective effects of recombinant adenovirus expressing glial cell line-derived neurotrophic factor (Ad-rGDNF) on the midbrain dopaminergic neurons against MPTP damage.
Methods:
Recombinant adenovirus carrying LacZ gene (Ad-LacZ) was injected into the murine striatum using the stereotaxic apparatus. The murine brains were perfused and dissected, then stained with X-Gal subsequently at 24 hours, 72 hours, 10 days and 30 days after injection. In another experiment, MPTP was injected subcutaneously into the mice 72 hours after the intrastriatal injection of Ad-rGDNF (1 x 10(8) pfu.ml-1). The murine striatum was removed to evaluate the content of DA, DOPAC and HVA by HPLC-ECD 1 week later.
Results:
X-Gal staining indicated the expression of LacZ gene 24 hours after injection of Ad-LacZ. Stronger expression was also detected at the end of 72 hours and 10 days, but decreased 30 days later. In addition, the LacZ gene expression was localized and detectable in both neurons and glial cells. There was no apparent cytotoxic effect induced by Ad-LacZ alone. In MPTP-treated mice, the content of dopamine decreased 80% compared to that in the saline-treated ones. The DA content of MPTP + Ad-rGDNF group on injected side was significantly higher than that of MPTP alone group and MPTP + Ad-LacZ group. Changes in the content of DOPAC and HVA followed the same pattern. The ratio of DOPAC/DA and HVA/DA on injected side also decreased significantly. However, no significant change was observed on uninjected side.
Conclusion:
The results suggested that recombinant adenoviral vector expressing rGDNF can protect DA neurons against MPTP injury.
