Non-specific inhibition of imipramine binding argues against an endogenous ligand

Abstract

We have attempted to characterize the product(s) inhibiting [3H]imipramine binding from rat cortical membranes. Fractions inhibiting [3H]imipramine binding were analysed by gas chromatography-mass spectrometry (GC-MS) and high performance liquid chromatography-mass spectrometry (HPLC-MS). Tissue extracts showed marked inhibitory activity (two different peaks); however, extracts containing no brain tissue (blanks) also exhibited inhibitory activity (one peak after HPLC pre-purification). The chemical composition of the compound that inhibited [3H]imipramine binding was identical in both cases (tissue and blank extracts) and consisted of zinc chelates, probably formed by the use of zinc sulfate during the extraction procedure. The second inhibitory peak in the tissue extracts was 5-hydroxytryptamine (serotonin, 5-HT). These results suggest that the inhibitory activity on [3H]imipramine binding attributed to a low-molecular weight putative endacoid is non-specific and due to ions introduced in the sample during purification.