The Investigation of the protective effect of hoat shock protein on neuronal death

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Gene transfer into rat brain: Rats (4, 8, or 12 weeks old) were anesthetized and fixed onto a stereotaxic operating table. A burr hole was opened and 20μg of plasmid DNA (pBK-CMV-HSP70) was injected into the hippocampus or striatum using a cannula. The injection time was changed from 10 sec to 10 min. Subsequently, a pair of stainless steel electrodes was inserted the region. Electric pulses were generated with a Square Electro Porator (CUY21) and were administered at 10 Hz. Electric pulses were 2ms in duration at 50, 100, or 150 V. However, at any condition, the sufficient gene transfer was not obtained. Only a couple of neurons showed β-galactosidase positive coincidentally.Transfection of HSP70 gene into PC12 cells: Human HSP75 cDNA was transfected using the pBK-CMV vector. Production of HSP70 protein was confirmed by Western blot analysis. The PC12 cells overexpressing HSP70 were exposed to hydrogen peroxide (50, 100, 150μM). MTT assay after 24h of exposure revealed the significant viability of HSP70-transfected PC12. The protective effect of HSP70 against oxidative injury was revealed.