Nicotinic acetylcholine receptors (nAChRs) and γ-aminobutyric acid type-A receptors (GABAARs) belong to the superfamily of ligand-gated ion channels. nAChRs are activated by the neurotransmitter acetylcholine and nicotine, mediating excitation. GABAARs are activated by GABA, conducting chloride, which mainly hyperpolarizes the membrane, thus inhibiting neurotransmission. The most abundant homomeric nAChR in the mammalian brain are the pentameric α7 nAChRs, which consist of five α7 subunits. Each subunit provides an orthosteric low affinity binding site for its endogenous ligand and a number of positive allosteric modulators (PAMs). Allosteric modulators of nAChRs have been implicated to be putative therapeutics in the pharmacological treatment of Alzheimer’s disease, schizophrenia and Parkinson’s disease. The GABAARs are also pentameric membrane proteins, possessing allosteric binding sites. PAMs of GABAARs are benzodiazepines and barbiturates, which are commonly used to treat the pathogenesis of epilepsy, neuropathic pain or developmental malfunctions. Hence, both, nACHRs and GABAARs have become potential targets for drugs, treating cognitive deficits, whereas PAMs get accredited a seminal role in the pharmacotherapy. Here we show the results of the target screen of the PAM PNU-120596 on nAChRs, and the PAM Diazepam on GABAARs performed on high throughput automated patch-clamp devices. Fast and accurate solution exchange (<10ms) and exposure times (<200ms) given by stacking solutions inside the pipette and rapid application to the cell enables reliable and repetitive activation of the GABAARs and even of fast desensitizing receptors such as nAChRs. Recently, Sitzia et al. (2011) also observed that physiological 37°C modulate the effect of PNU-120596 on nAChRs. To test, whether different temperatures affect PAMs of nAChRs and PAMs of GABAARs, we used a heated pipette to increase the temperature of the added solution and then rapidly applied to the cell. For both nAChRs and GABAARs, the PAM evoked currents significantly decreased with increasing temperature, supporting the idea of strong temperature dependence the allosteric modulation by PNU-102596 on nAChRs and by Diazepam on GABAARs. Taken together, we could reliably mimic the temperature dependence of PNU-120596. More importantly, we found for the first time an equal temperature dependence of GABAARs positive allosteric modulators.