Department of Biomedical Sciences, Korea University College of Medicine, #722 Moonsook Bldg., Inchon-Ro 73, Seongbuk-Gu, Seoul, 02841, South Korea.
College of Pharmacy, Chung-Ang University, Heukseok-Ro 84, Dongjak-Gu, Seoul, 06974, South Korea.
J Mol Neurosci. 2017 Dec;63(3-4):422-430. doi: 10.1007/s12031-017-0993-y. Epub 2017 Oct 31.
Primary sensory afferent neurons detect environmental and painful stimuli at their peripheral termini. A group of transient receptor potential ion channels (TRPs) are expressed in these neurons and constitute sensor molecules for the stimuli such as thermal, mechanical, and chemical insults. We examined whether a mouse sensory neuronal line, N18D3, shows the sensory TRP expressions and their functionality. In Ca imaging and electrophysiology with these cells, putative TRPV4-mediated responses were observed. TRPV4-specific sensory modalities including sensitivity to a specific agonist, hypotonicity, or an elevated temperature were reproduced in N18D3 cells. Electrophysiological and pharmacological profiles conformed to those from native TRPV4 of primarily cultured neurons. The TRPV4 expression in N18D3 was also confirmed by RT-PCR and Western blot analyses. Thus, N18D3 cells may represent TRPV4-expressing sensory neurons. Further, using this cell lines, we discovered a novel synthetic TRPV4-specific agonist, MLV-0901. These results suggest that N18D3 is a reliable cell line for functional and pharmacological TRPV4 assays. The chemical information from the novel agonist will contribute to TRPV4-targeting drug design.
初级感觉传入神经元在其外周末端检测环境和疼痛刺激。一组瞬时受体电位离子通道 (TRP) 在这些神经元中表达，构成了对热、机械和化学刺激等刺激的传感器分子。我们检查了小鼠感觉神经元系 N18D3 是否表现出感觉 TRP 的表达及其功能。在这些细胞的钙成像和电生理学中，观察到了推定的 TRPV4 介导的反应。N18D3 细胞再现了 TRPV4 特异性感觉模态，包括对特定激动剂、低渗或高温的敏感性。电生理学和药理学特征与原代培养神经元中的天然 TRPV4 一致。N18D3 中 TRPV4 的表达也通过 RT-PCR 和 Western blot 分析得到证实。因此，N18D3 细胞可能代表表达 TRPV4 的感觉神经元。此外，我们使用该细胞系发现了一种新型合成 TRPV4 特异性激动剂 MLV-0901。这些结果表明，N18D3 是用于功能性和药理学 TRPV4 测定的可靠细胞系。新型激动剂的化学信息将有助于 TRPV4 靶向药物设计。
