RatDISCO, a tissue clearing and immunolabelling protocol for large rat brains.
Craigie, K. J.; Till, S. M.; van de Gevel, B.; Vandrey, B.; Strangward, P.; Sieger, D.; Sulser, A. G.; Kind, P.; Rochefort, N. L.; Duguid, I.; Martinez-Gonzalez, C.
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RatDISCO is a simple, cost-effective, and reproducible tissue-clearing protocol optimised for immunolabeling in adult rat brains. It enables robust detection of diverse neuronal subtypes, glial populations, and vasculature, overcoming key limitations in antibody penetration and optical transparency. It is compatible with virally labelled-and transgenic mice, as well as human-derived brain organoids, highlighting its versatility across species and models. Functionally, it enables the detection of activity-dependent markers after behaviour, verified in a rat model of neurodevelopmental disorders, the Fragile X syndrome. These findings highlight RatDISCO as a broadly applicable tool for investigating neuroanatomical and functional alterations in rat models of disorders. HighlightsO_LIOptimised for immunolabeling in adult rat brains C_LIO_LIEnables detection of neurons, glial cells, and vasculature C_LIO_LIAchieves high tissue transparency without requiring specialised equipment. C_LIO_LICompatible with virally labelled mice and human brain organoids C_LIO_LIFacilitates mapping of activity-dependent markers in rat models of neurodevelopmental disorders C_LI
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