Center for Immunotherapy, Vaccines, and Virotherapy, Biodesign Institute, and School of Life Sciences, Arizona State University, Tempe, AZ, USA.
Methods Mol Biol. 2023;2597:89-104. doi: 10.1007/978-1-0716-2835-5_8.
The mechanisms underlying nervous system injury, such as spinal cord injury (SCI), traumatic brain injury (TBI), and peripheral nerve injury are complex and not well understood. Following acute tissue damage and cell death, inflammatory processes cause ongoing damage. Many factors regulate this inflammation, including factors that modulate chemokine expression. Serine proteases, including those of the thrombotic and thrombolytic pathways (e.g., thrombin, tPA, uPA) are upregulated during nervous system damage and can modulate the release and bioavailability of many chemokines. Virus-derived immunomodulators, such as Serp-1, a serine protease inhibitor (serpin), have protective effects by reducing inflammation and tissue damage. However, the precise mechanisms of Serp-1 neuroprotection are still being studied. Compartmentalized in vitro neuron culture systems, such as the Campenot trichamber, are useful for such mechanistic studies. This chapter provides a protocol for assembling and culturing rodent embryonic superior cervical ganglion (SCG) and dorsal root ganglion (DRG) neurons in Campenot trichambers, as well as instructive examples of the types of experiments enabled by these methods.
神经系统损伤的机制,如脊髓损伤 (SCI)、创伤性脑损伤 (TBI) 和周围神经损伤,非常复杂,目前还不完全清楚。在急性组织损伤和细胞死亡后,炎症过程会导致持续的损伤。许多因素调节这种炎症,包括调节趋化因子表达的因素。丝氨酸蛋白酶,包括血栓形成和纤溶途径中的那些(例如,凝血酶、tPA、uPA),在神经系统损伤过程中上调,并可调节许多趋化因子的释放和生物利用度。病毒衍生的免疫调节剂,如 Serp-1,一种丝氨酸蛋白酶抑制剂(丝氨酸蛋白酶抑制剂),通过减少炎症和组织损伤具有保护作用。然而,Serp-1 神经保护的确切机制仍在研究中。分区体外神经元培养系统,如 Campenot 三腔室,对于这些机制研究非常有用。本章提供了在 Campenot 三腔室中组装和培养啮齿动物胚胎颈上神经节 (SCG) 和背根神经节 (DRG) 神经元的方案,以及这些方法可实现的实验类型的示例。