Microfabricated Device to Record Axonal Conduction Under Pharmacological Treatment for Functional Evaluation of Axon Ion Channel.

Abstract

Neuronal networks are fundamental structures for information processing in the central nervous system. This processing function is severely impaired by abnormal axonal conduction from changes in functional ion channel expression. The evaluation of axonal conduction properties can be effective in the early diagnosis of information-processing abnormalities. However, little is known about functional ion channel expression in axons owing to lack of an appropriate method. In this study, we developed a device to measure changes in axonal conduction properties by selective pharmacological stimulation for the functional evaluation of Na channels expressed in axons. Axons of rat cortical neurons were guided across a pair of electrodes through microtunnel structures by employing surface patterning. The developed device detected more than 50 axons while recording for 10 min. The conduction delay along the axons decreased by 22.5% with neuron maturation. Tetrodotoxin and lidocaine (Na channel blockers) increased the conduction delay in a concentration-dependent manner depending on their working concentrations, indicating the effectiveness of the device. Finally, selective Na channel blockers for various Na channel subtypes were used. Phrixotoxin, a Nav1.2 blocker, markedly increased the conduction delay, suggesting that Nav1.2 is functionally expressed in the unmyelinated axons of the cerebral cortex. These results show that our device is feasible for the high-throughput functional evaluation of Na channel subtypes in axons. The results obtained can contribute to the understanding of the pathogenic mechanisms of neurological diseases that involve changes in the functional expression states of ion channels in axons.