We tested dinotefuran on hybrid SADβ2 nicotinic acetylcholine receptors (nAChRs) expressed in Xenopus laevis oocytes by electrophysiology to investigate how its unique structure influences agonist actions. Three structural variations (3-5) of dinotefuran were prepared by combining acyclic/cyclic nitroguanidine and tetrahydrofuran/acyclic ether to elucidate isosterism around the dinotefuran structure. Dinotefuran evoked dose-dependent inward currents in the hybrid nAChR with a pEC50 (–log EC50) of 3.65 ± 0.02 and an Imax (normalized maximum response) of 1.04 ± 0.03 (full agonist), while imidacloprid had a higher pEC50 (5.55 ± 0.06) but lower Imax (0.50 ± 0.02, partial agonist). Compound 3 (1-(3-tetrahydrofuranylmethyl)-2-nitroiminoimidazolidine) had a similar pEC50 (3.83 ± 0.06) to dinotefuran but much lower Imax (0.12 ± 0.01), and compounds 4 (1-(3-ethoxy-2-methyl)propyl-3-methyl-2-nitroguanidine) and 5 (1-(3-ethoxy-2-methyl)propyl-2-nitroiminoimidazolidine) showed no agonistic effects. The study demonstrated that dinotefuran has lower affinity but stronger channel-opening ability than imidacloprid for the SADβ2 hybrid nAChR, and it remains to be clarified how these electrophysiological features relate to its insecticidal potency and unique spectra in practice.