Induction of nitric oxide-dependent apoptosis in motor neurons by zinc- deficient superoxide dismutase

Science

Volumn 286, Issue 5449, 1999, Pages 2498-2500

  Estévez, Alvaro C ^a,e   Crow, John P ^a,d,e   Sampson, Jacinda B ^a,b,e   Reiter, Christopher ^a,b   Zhuang, Yingxin ^a   Richardson, Gloria J ^a   Tarpey, Margaret M ^a,e   Barbeito, Luis ^e,f   Beckman, Joseph S ^a,b,c,e  

^a UNIVERSITY OF ALABAMA AT BIRMINGHAM   (United States)

^b UNIVERSITY OF ALABAMA AT BIRMINGHAM   (United States)

^c University of Alabama at Birmingham   (United States)

^d University of Alabama at Birmingham   (United States)

^e UNIVERSITY OF ALABAMA AT BIRMINGHAM   (United States)

^f UNIVERSIDAD DE LA REPÚBLICA   (Uruguay)

Author keywords

[No Author keywords available]

Indexed keywords

COPPER ZINC SUPEROXIDE DISMUTASE; NITRIC OXIDE;

AMYOTROPHIC LATERAL SCLEROSIS; APOPTOSIS; ARTICLE; DISEASE CLASSIFICATION; ENZYME ACTIVITY; HUMAN; NERVE CELL NECROSIS; OXIDATION; PATHOGENESIS; PRIORITY JOURNAL; ZINC DEFICIENCY;

AMYOTROPHIC LATERAL SCLEROSIS; ANIMALS; APOPTOSIS; BRAIN-DERIVED NEUROTROPHIC FACTOR; CELLS, CULTURED; CHELATING AGENTS; COPPER; FLUORESCEINS; LIPOSOMES; MOTOR NEURONS; MUTATION; NITRATES; NITRIC OXIDE; NITRIC OXIDE SYNTHASE; NITRIC OXIDE SYNTHASE TYPE I; OXIDATION-REDUCTION; RATS; SUPEROXIDE DISMUTASE; SUPEROXIDES; ZINC;

EID: 0039251419     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.286.5449.2498     Document Type: Article

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35. SOD were expressed as described (6) and biotinylated with EZ-Link NHS-Biotin (Pierce, Rockford, IL). Liposomes were prepared by extrusion with SOD at 20 mg/0.1 ml [M. Hope, M. Bally, C. Webb, P. Cullis. Biochim. Biophys. Acta 812, 55 (1985); A. C. Estévez et al., Free Radical Biol. Med., in press]. Rat spinal motor neurons (200 cells per square centimeter) were cultured on polyornithine-laminin-coated 35-mm dishes in Neuro-basal medium (15) and exposed to liposomes (50 nmol lipid per square centimeter). The number of motor neurons with neuntes longer than four soma diameter treated with BDNF (100 pg/ml) at 24 hours was taken as 100% survival. Motor neuron death was prevented by caspase inhibitors (15) and showed nuclear condensation plus DNA fragmentation when visualized with the TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling) method, which together strongly suggests apoptosis.
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41. We thank C. Henderson (Developmental Biology Institute of Marseilles, Marseille, France) for many helpful discussions, R. Scott (Cephalon) for providing BDNF, and B. Mayer (Graz, Austria) for the neuronal NOS antibody. Supported by the ALS Association (J.S.B., A.G.E., and J.P.C) and the Public Health Service (grants R01 NS33291, RO1 NS36761, and RO1 HL58209 to J.S.B., R29 NS35871 to J.P.C, and K08 HL-03457 to M.M.T.).