Two phosphatidylethanol classes separated by thin layer chromatography are produced by phospholipase D in rat brain hippocampal slices.
Noradrenaline- and ionomycin-stimulated as well as basal phospholipase D activity from rat hippocampus produced, in the presence of ethanol, two different classes of [32P]phosphatidylethanol (designated I and II), which were separated by thin layer chromatography. Endogenous labeling experiments using 3H-fatty acids showed that two different classes of phosphatidylcholine, separated by two-dimensional TLC, one enriched with high incorporation of [3H]arachidonic acid (B) and the other with [3H]myristic acid (A), were the most likely sources for the two classes of phosphatidylethanol. Experiments where individual 32P-phospholipids extracted from [32P]Pi-labeled hippocampal slices were incubated with cabbage phospholipase D, in the presence of ethanol, showed that each class of [32P]phosphatidylcholine, i.e. A and B, produced a different band of [32P]phosphatidylethanol, with the same mobility in TLC as phosphatidylethanol II and I, respectively.
Sarri
E
E
Departament de Bioquimica i Biologia Molecular, Facultat de Medicina, Universitat Autònoma de Barcelona, Bellaterra, Spain.
Servitja
J M
JM
Picatoste
F
F
Claro
E
E
D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism; D001345:Autoradiography; D002855:Chromatography, Thin Layer; D000431:Ethanol / Q000378:metabolism; D005227:Fatty Acids / Q000032:analysis; D020404:Glycerophospholipids; D006624:Hippocampus / Q000201:enzymology*; D066298:In Vitro Techniques; D019814:Myristic Acid; D009227:Myristic Acids / Q000378:metabolism; D010710:Phosphates / Q000378:metabolism; D010712:Phosphatidic Acids / Q000302:isolation & purification / Q000378:metabolism*; D010713:Phosphatidylcholines / Q000302:isolation & purification / Q000378:metabolism; D010739:Phospholipase D / Q000378:metabolism*; D010761:Phosphorus Radioisotopes; D051381:Rats; D014316:Tritium
D005227:Fatty Acids; D020404:Glycerophospholipids; D009227:Myristic Acids; D010710:Phosphates; D010712:Phosphatidic Acids; D010713:Phosphatidylcholines; D010761:Phosphorus Radioisotopes; C051521:phosphatidylethanol; D019814:Myristic Acid; D014316:Tritium; D016718:Arachidonic Acid; D000431:Ethanol; D010739:Phospholipase D