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J Neurosci Methods. 1997 Apr 25;73(1):61-7.

Effect of neostigmine on concentration and extraction fraction of acetylcholine using quantitative microdialysis.

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Department of Chemistry, Emory University, Atlanta, GA 30322, USA.


A quantitative microdialysis method was used to determine the effect of local perfusion of 0, 100, 200, and 300 nM neostigmine (NEO) on acetylcholine (ACh) extracellular concentration and microdialysis extraction fraction (E(d)) in the striatum of the rat. Because of the efficiency of AChE, the inhibition of this enzyme is expected to result in a substantial increase in ACh levels and a decrease in the E(d) of ACh. The extracellular concentration of ACh increased linearly with increasing concentrations of NEO. The control ACh concentration was determined to be 18.4 +/- 11.8 nM (n = 10; mean +/- S.E.M.) The ACh extracellular concentration for the remaining groups was determined to be 173 +/- 14 nM (n = 5), 329 +/- 52.5 nM (n = 13), and 581 +/- 109 nM (n = 10) for the 100, 200, and 300 nM NEO groups, respectively. Perfusion with 300 nM NEO resulted in a significant reduction in the E(d) of ACh (64.5 +/- 3.5% vs. 43.6 +/- 7.5%, P < 0.05). In contrast to ACh, perfusion with 0, 1, and 10 microM hemicholinium-3, an inhibitor of high-affinity choline uptake, increased choline levels but did not affect the E(d) of choline. The effects on E(d) are consistent with E(d) being influenced by rapid clearance mechanisms.

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