Brar Harjot Kaur, Bassi Dikshalee, Ghosh-Roy Anindya
National Brain Research Centre, Manesar, Gurugram, India.
Bio Protoc. 2025 Apr 20;15(8):e5280. doi: 10.21769/BioProtoc.5280.
Research into nervous system injuries and regeneration has emerged as a crucial field of study. In many cases such as trauma or stroke, both axons and dendrites are equally damaged; however, studying injury and repair mechanisms in both neurite processes (axons and dendrites) of the same neuron has been challenging. Additionally, correlating the behavioral aspects of neuronal injury with anatomical regeneration is important for a better understanding of the functional rewiring process. Here, we describe protocols for injuring the dendrites and the axon of the PVD neuron of using a two-photon infrared (IR) femtosecond laser system, and subsequent imaging of injured neurites during the course of regeneration. Additionally, we describe the protocols for the behavioral study concerning the PVD neuron and their analysis, which can offer valuable insights. These assays can be implemented to assess the function of the pathways that play specific roles in dendrite vs. axon regeneration. Key features • Dendritic and axonal injury in PVD neurons using a two-photon microscope. • Behavioral analysis involving harsh touch response and proprioception following neurite injury. • Correlation of dendrite and axon regeneration events with behavioral parameters.
神经系统损伤与再生的研究已成为一个关键的研究领域。在许多情况下,如创伤或中风,轴突和树突都会受到同等程度的损伤;然而,研究同一神经元的两种神经突(轴突和树突)的损伤和修复机制一直具有挑战性。此外,将神经元损伤的行为方面与解剖学再生相关联,对于更好地理解功能重新布线过程很重要。在这里,我们描述了使用双光子红外(IR)飞秒激光系统损伤PVD神经元的树突和轴突的方案,以及在再生过程中对受损神经突进行后续成像的方案。此外,我们还描述了关于PVD神经元的行为研究及其分析的方案,这些方案可以提供有价值的见解。这些测定可用于评估在树突与轴突再生中起特定作用的通路的功能。关键特性 • 使用双光子显微镜对PVD神经元进行树突和轴突损伤。 • 神经突损伤后涉及强烈触摸反应和本体感觉的行为分析。 • 树突和轴突再生事件与行为参数的相关性。