Department of Medical Neuropharmacology, School of Pharmaceutical Sciences, Wakayama Medical University, Wakayama 640-8156, Japan.
Department of Clinical Pharmacology and Therapeutics, Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto 606-8507, Japan.
Cell Rep Methods. 2024 Aug 19;4(8):100835. doi: 10.1016/j.crmeth.2024.100835. Epub 2024 Aug 7.
We developed a rat dorsal root ganglion (DRG)-derived sensory nerve organotypic model by culturing DRG explants on an organoid culture device. With this method, a large number of organotypic cultures can be produced simultaneously with high reproducibility simply by seeding DRG explants derived from rat embryos. Unlike previous DRG explant models, this organotypic model consists of a ganglion and an axon bundle with myelinated A fibers, unmyelinated C fibers, and stereo-myelin-forming nodes of Ranvier. The model also exhibits Ca signaling in cell bodies in response to application of chemical stimuli to nerve terminals. Further, axonal transection increases the activating transcription factor 3 mRNA level in ganglia. Axons and myelin are shown to regenerate 14 days following transection. Our sensory organotypic model enables analysis of neuronal excitability in response to pain stimuli and tracking of morphological changes in the axon bundle over weeks.
我们通过在类器官培养装置上培养 DRG 外植体,开发了一种大鼠背根神经节 (DRG) 衍生的感觉神经器官型模型。通过这种方法,只需将源自大鼠胚胎的 DRG 外植体接种,就可以同时大量且具有高度重现性地产生大量器官型培养物。与以前的 DRG 外植体模型不同,这种器官型模型由神经节和带有髓鞘 A 纤维、无髓鞘 C 纤维和立体髓鞘形成的郎飞结的轴突束组成。该模型还表现出细胞体对神经末梢化学刺激的钙信号反应。此外,轴突横断增加了神经节中激活转录因子 3 mRNA 的水平。在横断后 14 天,轴突和髓鞘被证明可以再生。我们的感觉器官型模型能够分析神经元对疼痛刺激的兴奋性,并在数周内跟踪轴突束的形态变化。