title: Phospholipase B activity and organophosphorus compound toxicity in cultured neural cells.
abstract: Organophosphorus compounds (OP) such as phenyl saligenin phosphate (PSP) and mipafox (MPX) which cause delayed neuropathy, inhibit neuropathy target esterase (NTE), while OPs such as paraoxon (PXN) react more readily with acetylcholinesterase. In yeast and mammalian cell lines, NTE has been shown to have phospholipase B (PLB) activity which deacylates intracellular phosphatidylcholine to glycerophosphocholine (GroPCho) and can be detected by metabolic labeling with [(14)C]choline. Here we investigated PLB activity in primary cultures of mouse neural cells. In cortical and cerebellar granule neurons and astrocytes, [(14)C]GroPCho labeling was inhibited by PSP and MPX: phenyl dipentylphosphinate (PDPP), a non-neuropathic NTE inhibitor, was more potent, while PXN, was substantially less so. In all three cell types, conversion of [(14)C]phosphatidylcholine to [(14)C]GroPCho over 24 h was relatively small (2.3-14%). Consequently, even with >80% inhibition of [(14)C]GroPCho production, increased [(14)C]phosphatidylcholine was not detected. At concentrations of 1-10 microM, only PSP was cytotoxic to cortical and cerebellar granule neurons after 24-h exposure. Moreover, dramatic changes in glial cell morphology were induced by PSP, but not PDPP or MPX, with rapid (2-3 h) rounding up of astrocytes and of Schwann cells in cultures of dissociated mouse dorsal root ganglia. We conclude that PLB activity is present in a variety of cultured mouse neural cell types but that acute loss of this activity is not cytotoxic. Conversely, the rapid toxic effects of PSP in vitro suggest that a serine hydrolase distinct from NTE is required continuously by neurons and glia.
authors.affiliation: MRC Toxicology Unit, University of Leicester, LE1 9HN, UK.
authors.forename: David J
authors.initials: DJ
authors.lastname: Read
authors.forename: Lynda
authors.initials: L
authors.lastname: Langford
authors.forename: Helen R
authors.initials: HR
authors.lastname: Barbour
authors.forename: Philip J
authors.initials: PJ
authors.lastname: Forshaw
authors.forename: Paul
authors.initials: P
authors.lastname: Glynn
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology; D002540:Cerebral Cortex / Q000166:cytology; D008245:Lysophospholipase / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D015394:Molecular Structure; D009474:Neurons / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D009943:Organophosphorus Compounds / Q000737:chemistry / Q000633:toxicity*; D013329:Structure-Activity Relationship
chemical_list: D009943:Organophosphorus Compounds; D008245:Lysophospholipase