Neuronal nicotinic acetylcholine (AcCho) receptors composed of α7-subunits (α7-AcChoRs) are involved in many physiological activities. Nevertheless, very little is known about their single-channel characteristics. By using outside-out patch-clamp recordings from Xenopus oocytes expressing wild-type ( wt ) α7-AcChoRs, we identified two classes of channel conductance: a low conductance (γ L ) of 72 pS and a high one (γ H ) of 87 pS, with mean open-times (τ op ) of 0.6 ms. The same classes of conductances, but longer τ op (3 ms), were seen in experiments with chimeric α7 receptors in which the wtα7 extracellular C terminus was fused to the green fluorescent protein ( wt α7-GFP AcChoRs). In contrast, channels with three different conductances were gated by AcCho in oocytes expressing α7 receptors carrying a Leu-to-Thr 248 mutation ( mut α7) or oocytes expressing chimeric mut α7-GFP receptors. These conductance levels were significantly smaller, and their mean open-times were larger, than those of wt α7-AcChoRs. Interestingly, in the absence of AcCho, these oocytes showed single-channel openings of the same conductances, but shorter τ op , than those activated by AcCho. Accordingly, human homomeric wt α7 receptors open channels of high conductance and brief lifetime, and fusion to GFP lengthens their lifetime. In contrast, mut α7 receptors open channels of lower conductance and longer lifetime than those gated by wt α7-AcChoRs, and these parameters are not greatly altered by fusing the mut α7 to GFP. All this evidence shows that GFP-tagging can alter importantly receptor kinetics, a fact that has to be taken into account whenever tagged proteins are used to study their function.