Effects of Huperzine used as pre-treatment against soman-induced seizures.

Huperzine A (HUP), an alkaloid isolated from the Chinese club moss, Huperzia serrata is a reversible inhibitor of cholinesterases which crosses the blood-brain barrier and shows high specificity for acetylcholinesterase (AChE) and a prolonged biological half-life. We tested, in vivo, its efficiency in protecting cortical AChE from soman inhibition and preventing subsequent seizures. The release of acetylcholine (ACh) was also followed in the cortex of freely moving rats using microdialysis techniques. We previously found that soman-induced seizures occurred in rodents only when the cortical AChE inhibition was over 65% and when the increase of ACh level was over 200 times the baseline level. This was verified in the present study in control animals intoxicated by 1 LD50 of soman (90 microg/kg). Using the same dose of soman in rats pre-treated with 500 microg/kg of HUP, we observed that 93% of the animals survived and none of them had seizures. This dose of HUP reduced AChE inhibition to 54% and increase of ACh level to 230 times baseline value. HUP thus appears as a promising compound to protect subjects against organophosphorus intoxication.

Acetylcholine release from frontal cortex in the waking rat measured by microdialysis without acetylcholinesterase inhibitors: effects of diisopropylfluorophosphate.

Acetylcholine (ACh) release was measured in frontal cortex of awake quietly resting rats by microdialysis without using cholinesterase blockers in the perfusate. Resting release was 16.61 +/- 2.05 fmol/h (+/- S.E.M., n = 18). Injection of sublethal doses of the acetylcholinesterase blocker, diisopropylfluorophosphate produced dose-dependent increases in ACh release, reaching 79.9 fmol/h with a dose of 0.7-times the LD50. Although this irreversible inactivation of acetylcholinesterase increased ACh recovery to more than 700% of control values, levels of ACh in the perfusate never reached those seen in physostigmine-treated animals. The relationship between the amount of acetylcholinesterase inactivation and the quantity of ACh in the perfusate suggests that the extracellular ACh concentrations are controlled by simple enzyme kinetics. Within 2 h after enzyme inactivation, extracellular choline levels fell significantly, suggesting that ACh degradation by acetylcholinesterase plays an important role in regulating the amount of choline in the extracellular space.