Developmental, time-of-day, and stimulus-specific Golgi-Cox staining patterns detected in the mouse brain

Well-coordinated brain activity is a crucial driver of bodily functions and is refined by environmental input. Understanding the structure of brain areas that regulate various functions and how the environment affects the neural responses in the brain has been fundamental in advancing the field of neuroscience and medicine. The Golgi-Cox method is a histological approach that has allowed researchers to view neuronal structures with unmatched and detailed resolution, allowing for the comparison between non-diseased and diseased models, for instance. However, this method is known to stain neurons sparsely, which is useful for distinguishing structural components, but unpredictably, which is difficult for reproducibility and targeted studies. Here, we use three approaches to demonstrate a predictable pattern of cell staining using the Golgi-Cox method. We show that neuronal maturity, time of day, and response to environmental stimuli affect the number of cells stained by the Golgi-Cox method. Furthermore, we found low variability within each experimental group, which indicates staining reproducibility under controlled environments. Our study highlights important parameters for using the Golgi-Cox method and demonstrates its feasibility for broader application in answering neuroscience-based questions.