Enhancement of α7 nicotinic receptor (nAChR) function by positive allosteric modulators (PAMs) is a promising therapeutic strategy to improve cognitive deficits. PAMs have been classified only on the basis of their macroscopic effects as type I, which only enhance agonist-induced currents, and type II, which also decrease desensitization and reactivate desensitized nAChRs. To decipher the molecular basis underlying these distinct activities, we explored the effects on single-α7 channel currents of representative members of each type and of less characterized compounds. Our results reveal that all PAMs enhance open-channel lifetime and produce episodes of successive openings, thus indicating that both types affect α7 kinetics. Different PAM types show different sensitivity to temperature, suggesting different mechanisms of potentiation. By using a mutant α7 receptor that is insensitive to the prototype type II PAM (PNU-120596), we show that some though not all type I PAMs share the structural determinants of potentiation. Overall, our study provides novel information on α7 potentiation, which is key to the ongoing development of therapeutic compounds.

Ligand-gated ion channel kinetics were studied in mammalian transfected cells encoding adult mouse muscle acetylcholine (ACh) receptors. We measured macroscopic and single-channel currents using the outside-out and cell-attached patch-clamp configurations. Cultured cells were exposed to moderate intensity inhomogeneous static magnetic fields up to 180 mT and measurements were performed for temperatures ranging from 5 to 50 °C. We found no significant changes in ACh-elicited macroscopic or single-channel currents. We observed the expected dependence in current decay constants with temperature, but negligible magnetic field influence on the channel's kinetics.