Abstract:
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 Ca2+ 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.
摘要:
我们通过在类器官培养装置上培养DRG外植体,开发了大鼠背根神经节(DRG)衍生的感觉神经器官型模型。使用这种方法,只需接种来自大鼠胚胎的DRG外植体,就可以同时产生大量可重复性高的器官型培养物。与以前的DRG外植体模型不同,这个器官型模型由神经节和带有髓鞘A纤维的轴突束组成,无髓鞘C纤维,和Ranvier的立体髓鞘形成节点。该模型还表现出响应于对神经末梢的化学刺激的细胞体中的Ca2信号传导。Further,轴突横切增加神经节中激活转录因子3的mRNA水平。显示轴突和髓鞘在横切后14天再生。我们的感觉器官模型能够分析响应疼痛刺激的神经元兴奋性,并跟踪几周内轴突束的形态变化。