Synthesis of lipids in mouse brain cell cultures during development

Siegrist, H P; Bologa-Sandru, Liane; Burkart, Thomas; Wiesmann, Ulrich; Hofmann, K; Herschkowitz, N

doi:10.1002/jnr.490060304

Cited by 32 publications

(13 citation statements)

References 18 publications

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“…The lipid-insoluble residuum (containing mainly proteins) was solubilized bysonication in 1 WNaOH (quoted as 'protein fraction'). The radioactivity in the protein fraction was counted in an aliquot, and total proteins were determined by the method o f Lowry et al [32,33], Phospholipids, cholesterol, phosphatidylcthanolamines (PhE), cerebrosides and sulfatides were separated on Merck TLC plates (silica gel 60) using a mobile phase containing chloroform, methanol and water (130:56:9). The spots corresponding to the different lipid classes were collected, and the incorporated amounts o f radioactivity were determined.…”

Section: Determination O F Radioactivity Incorporated Into Lipids Andmentioning

confidence: 99%

“…The total lipid (TL) amount was determined by weigh ing the dry lipid extract [30][31][32], The lipids were then diluted in chloroform/methanol (2:1), quoted as 'lipid fraction', and the content of radioactivity was determined. The upper phase o f the lipid extraction (quoted as 'water-soluble fraction', containing precursor and gangliosides) was saved.…”

Section: Determination O F Radioactivity Incorporated Into Lipids Andmentioning

confidence: 99%

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N-Acetyl-<i>L</i>-Aspartate is a Major Source of Acetyl Groups for Lipid Synthesis during Rat Brain Development

1991

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The function of N-acetyl-L-aspartate (NAA), a predominant substance in the CNS, has not yet been determined. To investigate the possible function of NAA as a lipid precursor [¹⁴C]-N-acetyl-L-aspartate (NAA) or [¹⁴C]-acetate (AcA) was injected intracerebrally into 8, 15- and 22-day-old rats. These time points were selected because NAA concentration and the activity of the NAA synthetizing enzyme L-aspartate-N-acetyltransferase (ANAT) were low in 8-day-old rats, intermediate in 15-day-old rats and high in 22-day-old rats. During an incubation period of 4 h the radioactive acetyl group of NAA is incorporated into the lipid fraction in amounts of 42.9 to 65.7% of recovered total radioactivity, increasing with the age of the rats. In contrast, radioactivity incorporated from AcA is constant for all three ages. With NAA as precursor only 7.2–9.4% of the recovered total radioactivity is incorporated into the protein fraction. With AcA as precursor 27.0–18.1% of recovered radioactivity is incorporated into the protein fraction, the amounts decreasing with age. Taking into account that in vivo NAA concentration in the brain is much higher than the AcA concentration, NAA is clearly the more efficient precursor for lipid synthesis than AcA. Further, we compared NAA and AcA as lipid precursors by analyzing the radioactivity in single lipid fractions, expressed as normalized specific incorporation or normalized incorporation. The measured differences between NAA and AcA in normalized specific and normalized incorporation of acetyl groups imply that NAA is not simply degraded to AcA before incorporated into lipids. We conclude that NAA is a major source of acetyl groups for lipid synthesis during rat brain development.

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Section: Determination O F Radioactivity Incorporated Into Lipids Andmentioning

confidence: 99%

Section: Determination O F Radioactivity Incorporated Into Lipids Andmentioning

confidence: 99%

N-Acetyl-<i>L</i>-Aspartate is a Major Source of Acetyl Groups for Lipid Synthesis during Rat Brain Development

1991

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“…The lipid fraction of brain tissue, containing also the phospholipid PhE [7,13,22], was extracted in chloro form/methanol (2:1). The total lipid (TL) amount was determined by weighing the dry lipid extract.…”

Section: Determination O F Lipids and Pementioning

confidence: 99%

“…CST activity was determined by using 35-S phosphoadenos.ne-5-phosphosulfate as a sulfur donor [22,23], MBP content was determined in brain homogenates by radioimmunoassay according to the procedure of Bilrgisser [6].…”

Section: Determination O F Proteins and Enzyme A Dividesmentioning

confidence: 99%

Brain Damage and Recovery in Hyperphenylalaninemic Rats

Burri

Matthieu²,

Vandevelde³

et al. 1990

Dev Neurosci

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Rats were made hyperphenylalaninemic by injecting a mixture of alpha-methylphenylalanine and phenylalanine. Brain development was measured by biochemical, histological and 31-P nuclear magnetic resonance (NMR) methods. In 17-day-old hyperphenylalaninemic rats, brain myelinogenesis was disturbed. Compared to controls, test animals had lower body weights, brain weights, cerebrosides, sulfatides, myelin basic protein (MBP) and reduced cerebroside sulfotransferase (CST) and 2''3''-cyclic nucleotide-3''-phosphohydrolase (CNP) activities. No changes were found in total proteins, total lipids, total phospholipids, phosphatidylethanolamine and phosphorylethanolamine. In the brain of 17-day-old hyperphenylalaninemic rats no changes in phosphomonoesters, phosphodiester and phosphocreatine were found using in vivo 31-P NMR spectroscopy. Because body weights of hyperphenylalaninemic rats were significantly lower than those of controls, we compared them with undernourished rats. Undernourished rats had lower body weights, brain weights and CNP activity. No other changes were found. Therefore, we conclude that hyperphenylalaninemia per se and not undernutrition affected myelinogenesis in test animals. After treatment was discontinued, test rats recovered completely within 6 weeks with regard to biochemical and histological measurements; at 59 days they had normal body weights, cerebrosides, sulfatides, MBP, total proteins, total lipids, total phospholipids, phosphatidylethanolamine, phosphorylethanolamine and normal CST and CNP activities. Brain weights were significantly reduced.

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“…It has been used before to study utilization of glucose (33), ketones, and oleate (4). In the present work, experiments were performed when the cultures were at a developmental stage characterized by a high number of oligodendrocytes (3) and active synthesis of sulfatide, a major galactolipid of myelin (27). Galactose was studied in these brain cell cultures in order to test its suitability as a substitute for glucose, to elucidate its metabolic interaction with glucose, and to define its role in galactolipid formation.…”

mentioning

confidence: 99%