• Energy Research

We investigated the effect of phosphatidylethanol (PEt) on fluidity and membrane tolerance to the fluidization induced by ethanol as well as on the activity of two membrane-bound enzymes, Na+/K+ ATPase and 5'-nucleotidase. PEt was synthesized from 1,2-dimyristoylphosphatidylcholine and phosphatidylcholine from bovine brain and studies were performed to determine the optimal experimental conditions for the insertion of PEt in natural bilayers. The effects of PEt, evaluated by differential scanning calorimetry or fluorescence polarization techniques, were studied in model membranes made of synthetic phospholipids or made of total lipids extracted from rat brain crude mitochondrial fraction (P2 fraction) and from natural membranes (P2 fraction). The presence of PEt increased the fluidity of artificial as well of natural membranes, but tolerance to the addition of ethanol, displayed by dimyristoylphosphatidylcholine vesicles and by natural membranes containing PEt, was lacking in vesicles made of dimyristoylphosphatidylethanolamine and in artificial bilayers reconstituted from total P2 lipid extracts, suggesting an involvement of PC on PEt-induced ethanol resistance. Na+/K+ ATPase activity was enhanced by the addition of small amounts of ethanol (up to 50 mM) and progressively inhibited at higher concentrations, while 5'-nucleotidase was not affected up to 400 mM ethanol. The presence of PEt in the bilayer exerted the opposite effects on the two enzymes, reducing the Na+/K+ ATPase activation induced by ethanol and enhancing 5'-nucleotidase activity. The mechanisms of the PEt-induced modifications are discussed.

We studied the effects of a four week administration of low doses of ethanol on glycoconjugates of the synaptosomal and microsomal fraction prepared from the brain of rats aged 2 and 7 months. Synaptosomes were the more sensitive to ethanol treatment. Total lipid bound sialic acid and neutral glycolipid and glycoprotein content were significantly reduced only in the synaptosomal fraction, with greater differences in the younger age, while glycoprotein sialic acid was not affected. None of the above differences were statistically significant in the microsomal fraction. Ganglioside pattern was altered only in the 2 month rats, showing a reduction of GM1 and GM1a in the synaptosomal fraction and of GD1a in the microsomal fraction. UDP-Gal: asialo-mucin galactosyltransferase, UDP-Gal: GlcCer galactosyltransferase, and UDP-Gal: GM2 galactosyltransferase activities were decreased and could account for the observed modifications in glycoconjugate content and distribution.

The aim of this study was to investigate the lipid content and composition of rat cerebellar granule cells grown in the presence of ethanol (40, 55, or 80 mM) during in vitro differentiation. Quantitative analyses showed no effects of 40 mM ethanol, whereas a significant increase of total cholesterol was observed at 55 mM. Cells exposed to the highest ethanol dose (80 mM) were characterized by a higher sialidase activity, and by the modification of the ganglioside pattern and phospholipid fatty acid composition. The observed modifications were accompanied by changes of membrane anisotropy fluorescence assessed by the fluorescent probe 1,6-diphenyl-1,3,5-hexatriene.

Rats of two different ages (2 and 7 months) were treated with an ethanol-containing liquid diet for 24 days and change of the ceramide composition of gangliosides were studied in the brain synaptosomal, microsomal and myelin fractions. Greater differences were observed in the younger age, where ethanol treatment caused a significant increase of C20:1 LCB in GM1 ganglioside of synaptosomes and microsomes and in GD1a of myelin.