title: Beta-oxidation of [1-14C]palmitic acid by mouse astrocytes in primary culture: effects of agents implicated in the encephalopathy of Reye's syndrome.
abstract: beta-Oxidation of [1-14C]palmitic acid was examined in homogenates of astrocytes cultured from neonatal mouse brain. Under optimal reaction conditions (< or = 50 micrograms protein, 10 min at 37 degrees C), oxidation increased as a function of palmitate concentration (15 microM to 2 mM) and reached a maximum rate of 1.98 +/- 0.29 nmol/min/mg protein (mean +/- SEM) at 0.2 mM substrate. Eadie-Hofstee analysis of data from four experiments yielded apparent values for Vmax of 1.87 nmol/min/mg protein, and for Km, 35-40 microM. There were no dramatic changes in the oxidation rate in cells between 10 and 36 days in culture. During the 10-min assays, less than 0.05% of the radioactivity was converted to 14CO2 by the astrocytes; water-soluble products accounted for 1-2% of the total substrate added. Studies with KCN indicated that 60-70% of the total activity occurred in the mitochondria. We have been studying the structural and functional changes associated with the cerebral encephalopathy of Reye's syndrome (RS). Three-week-old astrocytes exposed to serum from RS children for the final 7 days of culture exhibited minor mitochondrial pleomorphism and had increased numbers of other intracellular organelles. Examination of the effects of agents implicated in RS indicated that oxidation of [1-14C]palmitate was not altered by Na+ salicylate (1-3 mM), but was inhibited by the industrial surfactant, Toximul MP-8 (> or = 10 micrograms/ml), 4-pentenoic acid (> or = 0.1 microM), or with 4 days' exposure to ammonia (10 nM). The latter treatment also resulted in an increase in protein synthesis, cell volume, and malondialdehyde formation. These results suggest that some of the "toxins" implicated in RS inhibit fatty-acid oxidation in the astrocytes and produce other lipid-related abnormalities that could be related to encephalopathy.
authors: Department of Physiology & Biophysics, Dalhousie University, Halifax, Nova Scotia, Canada.
authors: M G
authors: MG
authors: Murphy
authors: C
authors: C
authors: Jollimore
authors: J F
authors: JF
authors: Crocker
authors: H
authors: H
authors: Her
mesh_terms: D000641:Ammonia / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000378:metabolism*; D001834:Body Water / Q000378:metabolism; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D006801:Humans; D007223:Infant; D050356:Lipid Metabolism; D015227:Lipid Peroxidation / Q000502:physiology; D008315:Malondialdehyde / Q000378:metabolism; D051379:Mice; D010084:Oxidation-Reduction; D019308:Palmitic Acid; D010169:Palmitic Acids / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism; D012202:Reye Syndrome / Q000378:metabolism*; D014280:Triglycerides / Q000378:metabolism; D015262:Xenobiotics / Q000494:pharmacology
chemical_list: D010169:Palmitic Acids; D011506:Proteins; D014280:Triglycerides; D015262:Xenobiotics; D019308:Palmitic Acid; D008315:Malondialdehyde; D000641:Ammonia