Advancing Precision in Neurotoxicity analysis with Tissue Clearing and Automated Quantification

초록

Neurotoxicity research plays a crucial role in understanding the impact of chemicals and compounds on the nervous system, particularly neurons. Historically, histological analysis of brain tissue has been the primary method for elucidating neurotoxicity mechanisms and outcomes. However, traditional techniques often involve section-based histology, leading to structural distortions and challenges in obtaining precise and reproducible data. Recent advancements in tissue clearing methods have transformed this field by enabling researchers to examine intact, three-dimensional neural structures without traditional sectioning. This paradigm shift aligns with the "3R" principles of animal research, emphasizing the reduction, refinement, and replacement of experimental procedures to minimize harm and improve research outcomes. Our study introduces a comprehensive pipeline that combines tissue clearing, Fluoro-Jade staining, and AMaSiNe software for quantification, addressing the challenges of neurotoxicity assessment. We demonstrate the feasibility of this approach by evaluating the effects of neurotoxic chemicals, such as acrylamide, kainic acid, and 2,4-dinitrotoluol, on mouse brain tissue. This approach not only enhances the accuracy and efficiency of neurotoxicity analysis but also provides valuable spatial information, particularly critical for understanding the diverse physiological functions of different brain regions. In accordance with the 3R principles in animal experimentation, our findings underscore the potential of this method to advance neurotoxicity research, providing a more refined and efficient approach for evaluating the effects of neurotoxic compounds.

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