The effect of water soluble cyanotoxin(s) produced by two species of Anabaena on the release of acetylcholine from the peripheral cholinergic nervous system of the rat airway.

A water extract of the lyophilised fresh-water alga Anabaena flos-aquae enhanced substantially the release of [(3)H]acetylcholine ([(3)H]acetylcholine and [(3)H]choline) from cholinergic nerves of rat bronchi. Parallel experiments performed with the related species Anabaena lemmermannii did not demonstrate this effect. The effect on the release of [(3)H]acetylcholine by A. flos-aquae extract was concentration dependent. The A. flos-aquae induced [(3)H]acetylcholine release was not reduced by exposure to a low concentration of Ca(2+), but ω-conotoxin GVIA (1.0 μM), a blocker of N-type Ca(2+) channels reduced the release of [(3)H]acetylcholine induced by the A. flos-aquae extract. Addition of verapamil in a concentration (1.0 μM) specific for inhibition of L-type Ca(2+) channels had no effect on the neurotransmitter release. A reduction in the release was, moreover, observed with the intracellular Ca(2+) chelator BAPTA/AM (30 μM) and with the Na(+) channel blocker tetrodotoxin (3.0 μM). During patch-clamp studies of GH(4)C(1) neuronal cells, which have L- and T-type Ca(2+) channels, but no Na(+) channels, it was shown that a water extract of A. flos-aquae depolarised these cells and reduced, rather than enhanced, the influx of Ca(2+). Such an effect was not seen following exposure of GH(4)C(1) cells to water extracts of A. lemmermannii. In addition to its presynaptic activity, the water extract of A. flos-aquae showed an antimuscarinic effect by displacing [(3)H]QNB binding from muscarinic receptors in homogenates of rat bronchi. A similar but more potent effect was observed during experiments with water extract of A. lemmermannii. None of the respective water extracts showed any effects on cholinesterase activities in rat bronchial smooth muscle. The present observations suggest, therefore, that water extracts of A. flos-aquae may depolarise cells by activation of mono and divalent cation channels in cholinergic nerve cells. These channels are probably Na(+) channels and N-type, but not L- or T-type Ca(2+) channels. L- and T-type Ca(2+) channels were blocked in experiments with GH(4)C(1) cells and high concentrations of Ca(2+) channel blockers were necessary to reduce the effects of A. flos-aquae extract in cholinergic nerves in the airways. Furthermore, A. flos-aquae extract may also mobilise Ca(2+) from intracellular compartments. A. lemmermannii, on the other hand, does not contain components which alter mono and divalent cation-fluxes across cell membranes, but may rather have substances with more potent antagonistic effects on muscarinic cholinergic receptors than what is observed in experiments with A. flos-aquae.