title: L-2-chloropropionic acid inhibits glutamate and aspartate release from rat cerebellar slices but does not activate cerebellar NMDA receptors: implications for L-2-chloropropionic acid-induced neurotoxicity.
abstract: L-2-Chloropropionic acid (L-CPA), when orally administered at single high dose to rats produces a selective lesion in the cerebellum involving destruction of a high proportion of granule cells by a mechanism which involves N-methyl-D-aspartate (NMDA) receptors. Receptor binding studies demonstrated that L-CPA a had low affinity at the glutamate and glycine binding sites at NMDA receptors (530-660 microM), respectively, whereas L-CPA did not displace [3H]AMPA, [3H]NBQX or [3H]kainate from AMPA or kainate receptors. Whole cell-patch clamp experiments using cultured granule cells failed to demonstrate changes in membrane potential of cultured granule cells when either L-CPA (0.25 or 1 microM) was added alone to the bathing solution, or in combination with glycine (10 microM). Furthermore L-CPA did not alter the magnitude of the inward current produced by application of NMDA (100 microM)) to cultured granule cells, in the presence of glycine, as measured by patch clamp techniques. Experiments were also performed to discover whether L-CPA may alter the release of the excitatory amino acids from the cerebellum, which may then indirectly alter activity at glutamate receptors, leading to neuronal cell death. L-CPA (2 mM) did not affect either basal or stimulated (electrical or high potassium) endogenous aspartate release from superfused cerebellar slices nor did it alter the basal or stimulated release of [3H]aspartate from preloaded slices when introduced into the superfusion medium over 30 min. However, when cerebellar slices were preincubated with 2 mM L-CPA for 2 h at concentrations that are known to be neurotoxic to the brain in vivo, but not in vitro, the stimulated endogenous glutamate and aspartate net release was significantly attenuated, as compared to controls. Basal release was not significantly affected by the introduction of L-CPA-induced cerebellar neurotoxicity may be related to the inhibition of excitatory amino acid release from the cerebellum. In conclusion, although L-CPA does not appear to directly alter NMDA receptor activity the L-CPA-induced cerebellar neurotoxicity may be related to the inhibition of excitatory amino acid release from the cerebellum.
authors.affiliation: Neurotoxicology Research Group, ZENECA Central Toxicology Laboratory, ZENECA Pharmaceuticals, Alderley Park, Macclesfield, Cheshire, UK.
authors.forename: P S
authors.initials: PS
authors.lastname: Widdowson
authors.forename: I
authors.initials: I
authors.lastname: Briggs
authors.forename: R E
authors.initials: RE
authors.lastname: BoSmith
authors.forename: N C
authors.initials: NC
authors.lastname: Sturgess
authors.forename: A
authors.initials: A
authors.lastname: Rosbottom
authors.forename: J C
authors.initials: JC
authors.lastname: Smith
authors.forename: I
authors.initials: I
authors.lastname: Wyatt
mesh_terms: D000818:Animals; D001224:Aspartic Acid / Q000378:metabolism*; D001667:Binding, Competitive; D002478:Cells, Cultured; D002531:Cerebellum / Q000187:drug effects* / Q000378:metabolism; D004353:Drug Evaluation, Preclinical; D005260:Female; D018698:Glutamic Acid / Q000378:metabolism*; D006843:Hydrocarbons, Chlorinated; D066298:In Vitro Techniques; D008297:Male; D009498:Neurotoxins / Q000633:toxicity*; D018408:Patch-Clamp Techniques; D011422:Propionates / Q000633:toxicity*; D011869:Radioligand Assay; D051381:Rats; D016194:Receptors, N-Methyl-D-Aspartate / Q000187:drug effects* / Q000378:metabolism
chemical_list: D006843:Hydrocarbons, Chlorinated; D009498:Neurotoxins; D011422:Propionates; D016194:Receptors, N-Methyl-D-Aspartate; D001224:Aspartic Acid; D018698:Glutamic Acid; C021974:2-chloropropionic acid