Complementary chromatographic analysis of free diacylglycerols and potential glycerophospholipid precursors in human SH-SY5Y neuroblastoma cells following incubation with lithium chloride.
We performed detailed chromatographic analyses on the molecular species of the major glycerophospholipids (GPLs) and free sn-1,2-diacylglycerols (DAGs) from SH-SY5Y human neuroblastoma cells following incubation with or without LiCl. For this comparison the inositol, choline, ethanolamine and serine GPLs were dephosphorylated with phospholipase C and the released sn-1,2-diacylglycerols along with the DAGs were subjected to high-temperature GLC on polar and non-polar capillary columns as their trimethylsilyl and tert.-butyl-dimethylsilyl ethers. A 30-min incubation with 10 mM LiCl increased the total amount of human neuroblastoma DAGs by 32-58% (P < 0.05) to 2.6 pmol/micrograms cell protein. This was accompanied by a limited qualitative shift in the molecular species pattern, the most obvious of which was the increase (13%) in the major saturated-polyunsaturated molecular species and the ca. 46% increase in the minor 18:1-18:1 species over control levels. The DAGs originated mainly from the inositol GPLs (IGPLs), as indicated by the high levels of the characteristic 18:0-20:4n6 (18:0-20:3n9) species in both IGPLs and DAGs, and to a lesser extent from the choline GPLs (CGPLs), as indicated by the high proportion in CGPLs of the oligoenoic species, which were largely absent from IGPLs. Alkenylacylglycerols were not detected in DAGs, although they made up some 60% of the total ethanolamine GPLs (EGPLs). No significant changes in the molecular species composition of the cellular GPLs, including IGPLs, were detected after exposure to LiCl.
Liepkalns
V A
VA
Ralph Lowell Laboratories, Mclean Hospital, Belmont, MA.
Myher
J J
JJ
Kuksis
A
A
Leli
U
U
Freysz
N
N
Hauser
G
G
D001932:Brain Neoplasms / Q000737:chemistry*; D002845:Chromatography; D004075:Diglycerides / Q000032:analysis*; D005994:Glycerophosphates / Q000032:analysis*; D006801:Humans; D007202:Indicators and Reagents; D018021:Lithium Chloride / Q000494:pharmacology*; D009447:Neuroblastoma / Q000737:chemistry*; D017646:Organosilicon Compounds; D014297:Trimethylsilyl Compounds / Q000737:chemistry; D014407:Tumor Cells, Cultured
D004075:Diglycerides; D005994:Glycerophosphates; D007202:Indicators and Reagents; D017646:Organosilicon Compounds; D014297:Trimethylsilyl Compounds; C009660:t-butyldimethylsilyl compounds; D018021:Lithium Chloride