Hong-sheng Miao1, Lu-yang Yu, Guo-zhen Hui, Li-he Guo. 1. Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
Abstract
AIM: To evaluate the antiapoptotic effect of the A20 gene in primary hippocampal neurons both in vivo and in vitro. METHODS: Primary hippocampal neurons in embryonic day 18 (E18) rats were transfected with the A20 gene by using the new Nucleofector electroporation transfection method. We then examined, whether A20-neurons possessed anti-apoptotic abilities after TNF-alpha stimulation in vitro. A20-neurons and pcDNA3-neurons were transplanted into the penumbra of the brains of rats that had been subjected to 90-min of ischemia induced by left middle cerebral artery occlusion (MCAO). RESULTS: A20-neurons resisted TNF-alpha induced apoptosis in vitro. The apoptosis rate of neurons overexpressing A20 (28.46%+/-3.87%) was lower than that in neurons transfected with pcDNA3 (53.06%+/-5.36%). More A20-neurons survived in the penumbra both 3-d and 7-d after transplantation than did sham pcDNA3 neurons. CONCLUSION: The novel function of A20 may make it a potential targets for the gene therapy for neurological diseases.
AIM: To evaluate the antiapoptotic effect of the A20 gene in primary hippocampal neurons both in vivo and in vitro. METHODS: Primary hippocampal neurons in embryonic day 18 (E18) rats were transfected with the A20 gene by using the new Nucleofector electroporation transfection method. We then examined, whether A20-neurons possessed anti-apoptotic abilities after TNF-alpha stimulation in vitro. A20-neurons and pcDNA3-neurons were transplanted into the penumbra of the brains of rats that had been subjected to 90-min of ischemia induced by left middle cerebral artery occlusion (MCAO). RESULTS: A20-neurons resisted TNF-alpha induced apoptosis in vitro. The apoptosis rate of neurons overexpressing A20 (28.46%+/-3.87%) was lower than that in neurons transfected with pcDNA3 (53.06%+/-5.36%). More A20-neurons survived in the penumbra both 3-d and 7-d after transplantation than did sham pcDNA3 neurons. CONCLUSION: The novel function of A20 may make it a potential targets for the gene therapy for neurological diseases.
Authors: Anita B Hjelmeland; Qiulian Wu; Sarah Wickman; Christine Eyler; John Heddleston; Qing Shi; Justin D Lathia; Jennifer Macswords; Jeongwu Lee; Roger E McLendon; Jeremy N Rich Journal: PLoS Biol Date: 2010-02-23 Impact factor: 8.029