Finneran Matthew C, Rhamani Tara, Salioski Ismaël Valentin, Schmitd Ligia B, Passino Ryan, Johnson Craig N, Giger Roman J
Neuroscience Graduate Program, University of Michigan Medical School, Ann Arbor, MI, USA.
Cell and Developmental Biology, University of Michigan Medical School, Ann Arbor, MI, USA.
bioRxiv. 2025 Jul 3:2025.07.02.662816. doi: 10.1101/2025.07.02.662816.
Recent advances in experimental strategies that promote axon regeneration in adult mammals lay the foundation for future therapies. Reliable and unbiased quantification of regenerated axons is challenging, yet essential for comparing the efficacy of individual treatments and identification of most efficacious combinatorial therapies. Here, we introduce , a user-friendly and freely available software for the rapid quantification of regenerated axons in longitudinal nerve tissue sections. automatically identifies and traces regenerated axons, generating quantitative measurements that closely match conventional manual quantification but with significantly greater speed. Key features include length-dependent axon quantification at defined intervals from the injury site and normalization of axon density to nerve diameter to account for anatomical variability. To facilitate high-throughput analysis, the software includes an image queuing function. Additional features of allow quantification of a range of labeled cellular structures. As a proof of concept, we demonstrate accurate quantification of regenerated axons in the optic nerve, retinal ganglion cells density in retinal flat-mounts, and regenerated axon bundles in injured sciatic nerves. Collectively, we introduce a new platform that is expected to streamline and standardize regenerative outcome assessments across diverse experimental conditions and laboratories.
促进成年哺乳动物轴突再生的实验策略的最新进展为未来的治疗奠定了基础。对再生轴突进行可靠且无偏差的量化具有挑战性，但对于比较个体治疗的疗效以及确定最有效的联合治疗方法至关重要。在此，我们介绍了一种用户友好且免费可用的软件，用于快速量化纵向神经组织切片中的再生轴突。该软件能自动识别并追踪再生轴突，生成与传统手动量化结果紧密匹配但速度显著更快的定量测量结果。关键特性包括从损伤部位开始按特定间隔进行长度依赖性轴突量化，以及将轴突密度归一化至神经直径以考虑解剖学变异性。为便于高通量分析，该软件具备图像排队功能。该软件的其他特性还允许对一系列标记的细胞结构进行量化。作为概念验证，我们展示了对视神经中再生轴突、视网膜平铺标本中视网膜神经节细胞密度以及坐骨神经损伤后再生轴突束的准确量化。总体而言，我们引入了一个新平台，有望在不同实验条件和实验室中简化并标准化再生结果评估。
