[Biochemical characterization of the optic nerve in mice overexpressing the P53 gen. Oxidative stress assays].
OBJECTIVE
The tumour inhibitor p53 gene has the ability of triggering proliferation arrest and cellular death by apoptosis subsequent to several factors, among them oxidative stress. The p53 protein is a major regulator of gene expression. Using genetically manipulated mice carrying an extra copy of gene p53 (transgenic mice super p53) versus control mice, we have investigated the generation of reactive oxygen species and antioxidant activity in the optic nerve of mice in relation to p53 availability.


METHODS
We studied two groups of 12-month-old mice of the strain C57BL/6: 1) super p53 group (Sp53) and 2) wild-type control group (CG). Mice were anesthetized in ether atmosphere and the eyeball and retrobulbar optic nerves were excised, washed, soaked in PBS, and stored in liquid nitrogen at -85 degrees C until processing. Three-four optic nerves from the same group were placed in an eppendorf tube, homogenized and enzymatic-colorimetric methods used to determine oxidative and antioxidant activities and the nitric oxide synthesis.


RESULTS
A significant increase in free radical formation (via lipid peroxidation; p<0.001), antioxidant activity (p<0.001) and nitric oxide synthesis (p<0.001) was found in the optic nerves from transgenic super p53 mice compared to respective controls.


CONCLUSIONS
The presence of an extra copy of the p53 gene correlated with redox status in the mouse optic nerve. This transgenic mouse could be useful as an experimental model to study cell resistance to neurodegenerative processes in relation to oxidative stress and to apoptosis induction, such as glaucomatous optic neuropathy or age-related macular degeneration.
Gallego-Pinazo
R
R
Hospital Universitario La Fe, Valencia, Miembro de la Unidad de Investigación Oftalmológica Santiago Grisolía, Valencia, España. robertogallego@comv.es
Zanón-Moreno
V
V
Sanz
S
S
Andrés
V
V
Serrano
M
M
García-Cao
I
I
Pinazo-Durán
M D
MD
D000818:Animals; D005786:Gene Expression Regulation; D016158:Genes, p53; D051379:Mice; D008810:Mice, Inbred C57BL; D009900:Optic Nerve / Q000378:metabolism*; D018384:Oxidative Stress; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis* / Q000235:genetics
D016159:Tumor Suppressor Protein p53