Center for Rare Diseases Göttingen (ZSEG), Department of Pediatrics, University Medical Center Göttingen, Georg August University, 37075 Göttingen, Germany; Max-Planck-Institute for Multidisciplinary Sciences, 37075 Göttingen, Germany; Department of Neurology, Ökumenisches Hainich Klinikum, 99974 Mühlhausen, Germany.
Neurosurgical Medicine, Yavapai Regional Medical Group, Prescott, AZ 86301, USA.
Neurosci Lett. 2024 Oct 15;841:137959. doi: 10.1016/j.neulet.2024.137959. Epub 2024 Aug 31.
Understanding the sequence of cellular responses and their contributions to pathomorphogical changes in spinal white matter injuries is a prerequisite for developing efficient therapeutic strategies for spinal cord injury (SCI) as well as neurodegenerative and inflammatory diseases of the spinal cord such as amyotrophic lateral sclerosis and multiple sclerosis. We have developed several types of surgical procedures suitable for acute one-time and chronic recurrent in vivo multiphoton microscopy of spinal white matter [1]. Sophisticated surgical procedures were combined with transgenic mouse technology to image spinal tissue labeled with up to four fluorescent proteins (FPs) in axons, astrocytes, microglia, and blood vessels. To clearly separate the simultaneously excited FPs, spectral unmixing including iterative procedures was performed after imaging the diversely labeled spinal white matter with a custom-made 4-channel two-photon laser-scanning microscope. In our longitudinal multicellular studies of injured spinal white matter, we imaged axonal dynamics and invasion of microglia and astrocytes for a time course of over 200 days after SCI. Our methods offer ideal platforms for investigating acute and chronic cellular dynamics, cell-cell interactions, and metabolite fluctuations in health and disease as well as pharmacological manipulations in vivo.
了解细胞反应的顺序及其对脊髓白质损伤病理变化的贡献，是开发脊髓损伤 (SCI) 以及脊髓退行性和炎症性疾病（如肌萎缩侧索硬化症和多发性硬化症）有效治疗策略的前提。我们已经开发出几种适合急性一次性和慢性复发性体内多光子显微镜检查的脊髓白质的手术方法[1]。复杂的手术程序与转基因小鼠技术相结合，可对用多达四种荧光蛋白 (FP) 标记的脊髓组织进行成像，这些 FP 标记在轴突、星形胶质细胞、小胶质细胞和血管中。为了清楚地分离同时激发的 FP，在用定制的 4 通道双光子激光扫描显微镜对多样化标记的脊髓白质进行成像后，进行了包括迭代过程的光谱去混合。在我们对损伤的脊髓白质的纵向多细胞研究中，我们对 SCI 后超过 200 天的时间内轴突的动态和小胶质细胞和星形胶质细胞的入侵进行了成像。我们的方法为研究健康和疾病中的急性和慢性细胞动力学、细胞-细胞相互作用以及代谢物波动以及体内药理学操作提供了理想的平台。
