Characterization of two classes of benzodiazepine binding sites in Schistosoma mansoni

As we have recently shown that GABA should be considered a putative neurotransmitter in Schistosoma mansoni, the present work aimed to search for GABAA receptors in adult worms using [H-3]-flunitrazepam to label the allosteric benzodiazepine binding site which is classically present on GABAA receptor complexes. We detected a large population (B-max=8 center dot 25 +/- 1 center dot 1 pmol . mg protein(-1)) of high affinity (K-d = 33 center dot 6 +/- 1 center dot 5 nM) binding sites for flunitrazepam. These sites harboured a singular pharmacological modulation that does not fit well with a mammalian central benzodiazepine receptor, mainly due to avery high affinity for Ro5-4864 and a very low affinity for clonazepam. We also detected a second population of benzodiazepine binding sites labelled with high affinity (IC50 = 85 nM) by [H-3]-PK11195, a selective ligand of the mammalian peripheral benzodiazepine receptor. In conclusion, this work describes the pharmacological properties of a large population of central-like benzodiazepine receptors supporting their study as putative new targets for the development of anti-patasitic agents. We also describe, for the first time, the presence of peripheral benzodiazepine receptors in this parasite.