Peroxisomes were purified from the nervous tissue of 14-day-old rats by means of a Nycodenz gradient. Peroxisomal enzymes exhibited different sedimentation patterns: dihydroxyacetone phosphate acyl-transferase equilibrates at 1.142 g/ml together with the first peak of catalase; palmitoyl-CoA oxidase and D-amino acid oxidase activities are mainly recovered at 1.154 g/ml; the second peak of catalase is found at 1.175 g/ml. Morphological and semi-quantitative analyses of immunogold-labelled peroxisomes reveal profound heterogeneity of the particles. Very small (=0.2 microm diameter), electron dense vesicles containing catalase or thiolase, but devoid of other tested enzymes, are preferentially found in the light region, together with larger ( > 0.2 < 0.3 microm) and less electron dense palmitoyl-CoA oxidase-positive peroxisomes. At intermediate density (1.154 g/ml) peroxisomes of more uniform size (0.25-0.27 microm), containing palmitoyl-CoA oxidase or thiolase with or without catalase are preferentially found. This population extends toward the densest region of the gradient, where very large D-amino acid oxidase-containing peroxisomes are also found. In this region, smaller peroxisomes, often polymorphic, which are catalase- and thiolase-positive and D-amino acid oxidase/palmitoyl-CoA oxidase-negative, are also observed. The possibility that the heterogeneity of neural peroxisomes may reflect both cellular heterogeneity and ongoing peroxisomal biogenesis is discussed.
Presence of heterogeneous peroxisomal populations in the rat nervous tissue
CIMINI, Anna Maria;CRISTIANO L.;
1998-01-01
Abstract
Peroxisomes were purified from the nervous tissue of 14-day-old rats by means of a Nycodenz gradient. Peroxisomal enzymes exhibited different sedimentation patterns: dihydroxyacetone phosphate acyl-transferase equilibrates at 1.142 g/ml together with the first peak of catalase; palmitoyl-CoA oxidase and D-amino acid oxidase activities are mainly recovered at 1.154 g/ml; the second peak of catalase is found at 1.175 g/ml. Morphological and semi-quantitative analyses of immunogold-labelled peroxisomes reveal profound heterogeneity of the particles. Very small (=0.2 microm diameter), electron dense vesicles containing catalase or thiolase, but devoid of other tested enzymes, are preferentially found in the light region, together with larger ( > 0.2 < 0.3 microm) and less electron dense palmitoyl-CoA oxidase-positive peroxisomes. At intermediate density (1.154 g/ml) peroxisomes of more uniform size (0.25-0.27 microm), containing palmitoyl-CoA oxidase or thiolase with or without catalase are preferentially found. This population extends toward the densest region of the gradient, where very large D-amino acid oxidase-containing peroxisomes are also found. In this region, smaller peroxisomes, often polymorphic, which are catalase- and thiolase-positive and D-amino acid oxidase/palmitoyl-CoA oxidase-negative, are also observed. The possibility that the heterogeneity of neural peroxisomes may reflect both cellular heterogeneity and ongoing peroxisomal biogenesis is discussed.Pubblicazioni consigliate
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