Full
Outline
Voltage-gated sodium channels (VGSCs, NaV) are essential for the generation and conduction of action potentials in excitable cells. The NaV1.8 subtype is predominantly expressed in sensory neurons, particularly in small-diameter neurons of the dorsal root ganglia (DRG), and plays a critical role in the signal transduction of chronic, neuropathic, and inflammatory pain. Due to its tetrodotoxin-resistant properties and electrophysiological characteristics of slow activation and inactivation, NaV1.8 is a key target for analgesic drug development, facilitating repeated neuronal firing. Consequently, establishing an in vitro drug screening model based on NaV1.8 is crucial for identifying lead compounds with analgesic potential. However, heterologous expression of NaV1.8 remains challenging, primarily due to low expression levels in non-neuronal cell lines, which hinders effective drug screening. In this study, we established an in vitro expression model of rat NaV1.8 using Xenopus laevis oocytes as the expression system. Channel property was characterized using the two electrode voltage clamp (TEVC) technique, providing a platform for screening active compounds. Quantitative polymerase chain reaction confirmed that rat NaV1.8 is predominantly expressed in the DRG and trigeminal ganglia (Guangxi University Ethics Committee Review and approval number: GXU-2022-159). We cloned the full-length NaV1.8 gene and constructed two chimeric variants, NaV1.8-N and NaV1.8-C. Following in vitro transcription of the corresponding capped RNA, it was injected into Xenopus laevis oocytes for heterologous expression. TEVC recordings were used to detect NaV1.8 and chimera expression, and channel activity was evaluated through I-V curves, steady-state activation curves, and the specific NaV1.8 antagonists A-803467 and VX-548. The chimera NaV1.8-C resolved the problem in stable expression of NaV1.8 in vitro and exhibited similar pharmacological properties with native NaV1.8, offering a valuable tool for future screening of compounds targeting voltage-gated sodium channel.
218
98
| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
关闭全屏
No. 86 Xueyuan South Road, Haidian District, Beijing
010-62662699(Operation and Technical Support)
qkjq@cast.org.cn
Copyright © 2025 China Association for Science and Technology. All rights reserved. For all open access content, the relevant licensing terms apply.
Sponsored by the Office of the Leading Group for Cybersecurity and Informatization of CAST, and supported by Science and Technology Review Publishing House
