Ultrastructural pathology and cytochemical investigations of L-2-chloropropionic acid-induced neurointoxication of the rat cerebellum.
The objectives of the studies described were to assess the ultrastructural neuropathology, blood-brain barrier (BBB) integrity and calcium status of the cerebellum of rats following a single dose of 750 mg.kg-1 L-2-chloropropionic acid (L-2-CPA). The first indications of intoxication appeared at 36 h when condensation of many granule cells associated with Purkinje cell degeneration and marked astroglial swelling were observed. Some electron-lucent granule cells were also noted lying amongst these condensed forms. Condensed granule cells had swollen, electron-lucent mitochondria, dilated Golgi apparatus and nuclear crenation. Occasionally, areas of the granule cell necrosis were also present at this time. Granule cell condensation probably represents a preliminary and irreversible stage in an excitotoxic process that leads to necrosis. At 48 and 72 h, most granule cells were necrotic, and occasionally, extravasation of both erythrocytes and leucocytes into the expanded extravascular space was observed. Evaluation of the BBB by ultrastructural cytochemical visualisation of horseradish peroxidase injected i.v. 2 min before killing by perfusion fixation showed substantial leakage. At 36 h post-dose, ultrastructural calcium localisation using oxalate/pyroantimonate precipitation demonstrated a substantial increase in calcium pyroantimonate precipitate in mitochondria and other membranous cytoplasmic organelles (especially the Golgi apparatus) in condensed granule cells, but with little in their nuclei. However, their immediate neighbours (of ostensibly normal ultrastructural appearances) contained greater amounts of intranuclear precipitate. Swollen astroglial cells (especially the Bergmann glia) contained considerable quantities of precipitate. A possible excitotoxic mechanism via L-2-CPA-induced NMDA receptor agonism leading to overwhelming calcium influx and disruption of cellular calcium homeostasis is proposed.
Jones
H B
HB
Safety and Medicines Group, Zeneca Pharmaceuticals, Macclesfield, Cheshire, UK.
Jenkins
C R
CR
Bowdler
A L
AL
Simpson
M G
MG
Lock
E A
EA
D000818:Animals; D002531:Cerebellum / Q000187:drug effects* / Q000648:ultrastructure*; D006843:Hydrocarbons, Chlorinated; D008297:Male; D008854:Microscopy, Electron; D009498:Neurotoxins / Q000633:toxicity*; D011422:Propionates / Q000633:toxicity*; D051381:Rats; D017208:Rats, Wistar
D006843:Hydrocarbons, Chlorinated; D009498:Neurotoxins; D011422:Propionates; C021974:2-chloropropionic acid