Long Keyao, Xu Wanyue, Miao Xun, Wang Su, Rui Menglong
School of Life Science and Technology, the Key Laboratory of Developmental Genes and Human Disease, Southeast University, Nanjing, China.
Bio Protoc. 2025 Jul 5;15(13):e5367. doi: 10.21769/BioProtoc.5367.
Over the lifespan of an individual, brain function requires adjustments in response to environmental changes and learning experiences. During early development, neurons overproduce neurite branches, and neuronal pruning removes the unnecessary neurite branches to make a more accurate neural circuit. motoneurons prune their intermediate axon bundles rather than the terminal neuromuscular junction (NMJ) by degeneration, which provides a unique advantage for studying axon pruning. The pruning process of motor axon bundles can be directly analyzed by real-time imaging, and this protocol provides a straightforward method for monitoring the developmental process of motor neurons using live cell imaging. Key features • Long-range projecting axon bundles of motor neurons extending from soma on the ventral nerve cord (VNC) undergo degeneration rather than retraction during metamorphosis. • The pruning process of motor axon bundles can be directly observed by real-time live-cell imaging. • The complete clearance of axon bundles occurs approximately 22 h after pupal formation (22 h APF). • Mushroom body (MB) γ neuron axon pruning regulatory genes are conserved for motor neurons.
在个体的整个生命周期中，大脑功能需要根据环境变化和学习经历进行调整。在早期发育过程中，神经元会过度产生神经突分支，而神经元修剪会去除不必要的神经突分支，以形成更精确的神经回路。运动神经元通过退化来修剪其中间轴突束，而不是终端神经肌肉接头（NMJ），这为研究轴突修剪提供了独特的优势。运动轴突束的修剪过程可以通过实时成像直接分析，本方案提供了一种使用活细胞成像监测运动神经元发育过程的直接方法。关键特征 • 从腹侧神经索（VNC）上的胞体延伸出的运动神经元的长距离投射轴突束在变态过程中发生退化而不是回缩。 • 运动轴突束的修剪过程可以通过实时活细胞成像直接观察到。 • 轴突束在蛹形成后约22小时（22 h APF）完全清除。 • 蘑菇体（MB）γ神经元轴突修剪调节基因在运动神经元中是保守的。
