Antiapoptotic effect both in vivo and in vitro of A20 gene when transfected into rat hippocampal neurons.
OBJECTIVE
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.


CONCLUSIONS
The novel function of A20 may make it a potential targets for the gene therapy for neurological diseases.
Miao
Hong-sheng
HS
Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
Yu
Lu-yang
LY
Hui
Guo-zhen
GZ
Guo
Li-he
LH
D000818:Animals; D017209:Apoptosis; D002478:Cells, Cultured; D015870:Gene Expression; D006624:Hippocampus / Q000166:cytology / Q000378:metabolism*; D020244:Infarction, Middle Cerebral Artery / Q000473:pathology* / Q000601:surgery; D008297:Male; D009474:Neurons / Q000166:cytology / Q000378:metabolism / Q000637:transplantation; D011506:Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D011506:Proteins; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha