N-methyl-D-aspartate and non-N-methyl-D-aspartate receptors mediate seizures and CA1 hippocampal damage induced by dendrotoxin-K in rats.
The epileptogenic and neurodegenerative effects of dendrotoxin K, from Dendroaspis polylepis, a specific blocker of a non-inactivating, voltage-sensitive K+ channel, were studied after focal injection into one dorsal hippocampus in rats. Administration of 35 pmol dendrotoxin K elicited motor seizures and bilateral electrocortical discharges after a latent period (5.3 +/- 2.1 min), in all of the treated animals (n = 6). At 24 h, histological examination of brain (n = 5) coronal sections (10 microns; n = 6 per brain) detected bilateral damage to the hippocampal formation which extended 300 microns rostral and caudal to the injection tract. Quantitation of the damage revealed significant bilateral neuronal cell loss in the CA1 and CA4 pyramidal cell layer relative to the corresponding brain regions of rats (n = 3) injected with bovine serum albumin (105 pmol), which per se was ineffective in all respects. Dendrotoxin K (35 pmol) also caused a significant loss of CA3 pyramidal neurons and dentate gyrus granule cells ipsilateral to the site of toxin injection. In one out of six rats, a lower dose (3.5 pmol) of dendrotoxin K produced convulsive behaviour and electrocortical seizures but after a longer latency and these were accompanied by significant neuronal loss in the CA1, CA3 and CA4 pyramidal cell layer ipsilateral to the injected side. The lowest dose (0.35 pmol; n = 6 rats) of dendrotoxin K used failed to induce seizures and did not cause hippocampal damage (n = 6 rats). Systemic (i.p.) treatment with dizocilpine maleate (3 mg/kg) or LY 274614 (5 mg/kg i.p.), two N-methyl-D-aspartate receptor antagonists (given 15 min beforehand), prevented dendrotoxin K (35 pmol)-induced motor seizures and electrocortical epileptogenic discharges in 100% of the animals (n = 6 per group) treated. Similarly, these antagonists minimized the damage typically produced in the rat hippocampus, with no significant neuronal loss being observed. By contrast, NBQX (30 mg/kg, i.p. given 15 min previously), a non-N-methyl-D-aspartate antagonist, failed to prevent seizures normally evoked by dendrotoxin K (35 pmol; n = 6 rats); also, this treatment was unable to abolish CA1 pyramidal cell loss but minimized the loss in hippocampal sectors distant to the site of dendrotoxin K injection. However, complete protection against motor and electrocortical seizures and hippocampal damage was afforded by GYKI 52466 (10 mg/kg i.p.; n = 6 rats), a more effective non-N-methyl-D-aspartate receptor antagonist. These findings differ from the reported lack of protection by N-methyl-D-aspartate and non-N-methyl-D-aspartate receptor antagonists to rats receiving intra-hippocampal injection of alpha-dendrotoxin; this difference may stem from the ability of alpha-dendrotoxin to block predominantly a slowly inactivating K+ current whereas dendrotoxin K inhibits a non inactivating variant. In conclusion, the present data on dendrotoxin K, together with the previously described pattern of neurotoxicity for alpha-dendrotoxin, show that these homologues act via different mechanisms and, thus, can be used effectively as complementary tools to study seizures and neuronal cell death.
Bagetta
G
G
Department of Neuroscience, University of Cagliari, Italy.
Iannone
M
M
Palma
E
E
Nisticò
G
G
Dolly
J O
JO
D000818:Animals; D016923:Cell Death; D004546:Elapid Venoms / Q000494:pharmacology*; D006624:Hippocampus / Q000187:drug effects* / Q000473:pathology*; D008297:Male; D015221:Potassium Channels / Q000187:drug effects; D051381:Rats; D017208:Rats, Wistar; D016194:Receptors, N-Methyl-D-Aspartate / Q000187:drug effects*; D012640:Seizures / Q000473:pathology*; D013997:Time Factors
D004546:Elapid Venoms; D015221:Potassium Channels; D016194:Receptors, N-Methyl-D-Aspartate; C020273:dendrotoxin