Hawker Brooke, Connor Bronwen, McCaughey-Chapman Amy
Department of Pharmacology and Clinical Pharmacology, Centre for Brain Research, School of Medical Sciences, Faculty of Medical and Health Sciences, University of Auckland, Auckland, New Zealand.
Front Cell Neurosci. 2025 May 22;19:1609806. doi: 10.3389/fncel.2025.1609806. eCollection 2025.
The development of organotypic slice cultures of central nervous system (CNS) tissues has bridged the gap between simple in vitro cell cultures and complex in vivo whole animal studies. Organotypic brain slice cultures are a useful tool to study neurological disease, providing a more complex 3-dimensional system than standard 2-dimensional in vitro cell culture. In particular, organotypic brain slice cultures provide an excellent model to study the processes of demyelination and remyelination associated with neurological disease and injury. However, organotypic brain slice cultures are typically generated using coronal sectioning or regionspecific hippocampal or cerebellar tissue. We have previously reported the ability to generate sagittal organotypic brain slice cultures, allowing us to investigate the anterior-to-posterior integrity of the corpus callosum during demyelination and remyelination processes. To extend our sagittal organotypic brain slice culture model, this study compares the ability for two common demyelinating agents, cuprizone (CPZ) or lysolecithin (LPC), to induce demyelination of the corpus callosum.
Rat brain sagittal organotypic slice cultures were generated with clear visualization of the corpus callosum and treated either with CPZ (1 mM) or LPC (0.5 mg/mL).
We demonstrate that CPZ treatment induces acute demyelination followed by endogenous remyelination 1-week post-treatment. Conversely, we show that LPC treatment results in prolonged demyelination of the corpus callosum that is maintained 5 weeks post-treatment and is associated with an acute astroglia response.
Overall, this study demonstrates the use of CPZ and LPC to model either acute or prolonged demyelination in a sagittal organotypic brain slice culture system. These models provide a platform for studying acute and chronic demyelination and for testing new therapeutic approaches aimed at enhancing remyelination prior to conducting in vivo experiments.
中枢神经系统（CNS）组织的器官型切片培养的发展弥合了简单体外细胞培养与复杂体内全动物研究之间的差距。器官型脑切片培养是研究神经疾病的有用工具，提供了比标准二维体外细胞培养更复杂的三维系统。特别是，器官型脑切片培养为研究与神经疾病和损伤相关的脱髓鞘和再髓鞘化过程提供了一个极好的模型。然而，器官型脑切片培养通常使用冠状切片或特定区域的海马或小脑组织来生成。我们之前报道了生成矢状器官型脑切片培养的能力，这使我们能够在脱髓鞘和再髓鞘化过程中研究胼胝体从前到后的完整性。为了扩展我们的矢状器官型脑切片培养模型，本研究比较了两种常见脱髓鞘剂，铜螯合剂（CPZ）或溶血卵磷脂（LPC），诱导胼胝体脱髓鞘的能力。
生成大鼠脑矢状器官型切片培养，清晰显示胼胝体，并用CPZ（1 mM）或LPC（0.5 mg/mL）处理。
我们证明CPZ处理诱导急性脱髓鞘，随后在处理后1周内发生内源性再髓鞘化。相反，我们表明LPC处理导致胼胝体长期脱髓鞘，在处理后5周仍持续存在，并伴有急性星形胶质细胞反应。
总体而言，本研究证明了使用CPZ和LPC在矢状器官型脑切片培养系统中模拟急性或长期脱髓鞘。这些模型为研究急性和慢性脱髓鞘以及在进行体内实验之前测试旨在增强再髓鞘化的新治疗方法提供了一个平台。
