tailieunhanh - Báo cáo khoa học: Adaptation to G93Asuperoxide dismutase 1 in a motor neuron cell line model of amyotrophic lateral sclerosis The role of glutathione

Motor neuron degeneration in amyotrophic lateral sclerosis involves oxida-tive damage. Glutathione (GSH) is critical as an antioxidant and a redox modulator. We used a motor neuronal cell line (NSC-34) to investigate whether wild-type and familial amyotrophic lateral sclerosis-linked G93A mutant Cu,Zn superoxide dismutase (wt⁄G93ASOD1) modified the GSH pool and glutamate cysteine ligase (GCL), the rate-limiting enzyme for GSH synthesis. | ỊFEBS Journal Adaptation to G93Asuperoxide dismutase 1 in a motor neuron cell line model of amyotrophic lateral sclerosis The role of glutathione Silvia Tartari1 Giuseppina D Alessandro1 Elisabetta Babetto1 Milena Rizzardini1 Laura Conforti2 and Lavinia Cantoni1 1 Department of Molecular Biochemistry and Pharmacology Istituto di Ricerche Farmacologiche Mario Negri Milan Italy 2 Department of Neuroscience Istituto di Ricerche Farmacologiche Mario Negri Milan Italy Keywords amyotrophic lateral sclerosis Cu Zn superoxide dismutase glutamate cysteine ligase glutathione motor neuron Correspondence L. Cantoni Laboratory of Molecular Pathology Department of Molecular Biochemistry and Pharmacology Istituto di Ricerche Farmacologiche Mario Negri Via G. La Masa 19 20156 Milan Italy Fax 39 02 354 6277 Tel 39 02 3901 4423 E-mail cantoni@ Present address Babraham Institute Cambridge UK Received 22 December 2008 revised 17 February 2009 accepted 18 March 2009 doi Motor neuron degeneration in amyotrophic lateral sclerosis involves oxidative damage. Glutathione GSH is critical as an antioxidant and a redox modulator. We used a motor neuronal cell line NSC-34 to investigate whether wild-type and familial amyotrophic lateral sclerosis-linked G93A mutant Cu Zn superoxide dismutase wt G93ASOD1 modified the GSH pool and glutamate cysteine ligase GCL the rate-limiting enzyme for GSH synthesis. We studied the effect of various G93ASOD1 levels and exposure times. Mutant Cu Zn superoxide dismutase induced an adaptive process involving the upregulation of GSH synthesis even at very low expression levels. However cells with a high level of G93ASOD1 cultured for 10 weeks showed GSH depletion and a decrease in expression of the modulatory subunit of GCL. These cells also had lower levels of GSH and GCL activity was not induced after treatment with the pro-oxidant tertbutylhydroquinone. Cells with a low level of G93ASOD1 maintained higher GSH levels