Amblyopia, a neurodevelopmental visual disorder characterized by impaired stereoacuity, is commonly modeled in animals using monocular deprivation (MD) during a critical period of visual development. Despite extensive research at the synaptic, cellular and circuit levels of analysis, reliable behavioral assays to study stereoscopic deficits in mice are limited. This study aimed to characterize the Visual Cliff Assay (VCA) and the Pole Descent Cliff Task (PDCT) in mice, and to evaluate their utility in detecting binocular dysfunction. Using these assays, we investigated the impact of clinically relevant manipulations of binocular vision, including monocular occlusion, pupillary dilation, and amblyopia induced by long-term MD. Our findings reveal that optimal performance in both the VCA and PDCT are dependent on balanced binocular input. However, deficits after MD in the VCA exhibited relatively small effect sizes (7-14%), requiring large sample sizes for statistical comparisons. In contrast, the PDCT demonstrated larger effect sizes (43-61%), allowing for reliable detection of binocular dysfunction with a smaller sample size. Both assays were validated using multiple monocular manipulations relevant to clinical paradigms, with the PDCT emerging as the preferred assay for detecting deficits in stereoscopic depth perception in mice. These findings provide a robust framework for using the VCA and PDCT in mechanistic and therapeutic studies in mice, offering insights into the neural mechanisms of binocular vision and potential interventions for amblyopia.
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