The elevated plus-maze is a widely employed behavioural assay for anxiolytic and anxiogenic agents in rodents. Among laboratories, however, the apparatus often differs between the use of transparent and opaque walls. This inconsistency may represent an unnecessary confound in the literature when comparing results. Here, we directly contrasted the two designs with respect to (1) mouse strain differences (C57BL/6, 129/Sv, and C3H/He) and (2) sensitivity to diazepam (0.5, 1.0, and 1.5 mg/kg, intraperitoneal). Both mazes yielded similar results indicating that 129/Sv mice were more anxious than C57BL/6 or C3H mice, with the transparent elevated plus-maze tending to encourage open-arm exploration in all three strains. Next, we examined the effect of diazepam in the 129/Sv strain across the two mazes. Systemic diazepam at 1.5 mg/kg led to increased percentage time spent in the open arms in both elevated plus-mazes; the drug was ineffective in both elevated plus-mazes at a dose of 0.5 mg/kg. Although our results revealed little practical difference between the two mazes in terms of their ability to detect differences in anxiety-related behaviour, the baseline difference in open-arm exploration between the two elevated plus-mazes suggests that the transparent design may be more congenial for the detection of anxiogenic manipulations, and the opaque maze for anxiolytic manipulations.