In Situ Investigation of Intercellular Signal Transduction Based on Detection of Extracellular pH and ROS by Scanning Electrochemical Microscopy

Intercellular signal transduction plays an important role in the regulation of biological activities. Herein, a Transwell chamber-based two-layer device combined with scanning electrochemical microscopy (SECM) technology has been proposed for in situ investigation of intercellular signal transduction. The cells in the device were cultured on two layers: the lower layer was for signaling cells, and the upper layer was for signal-receiving cells. The extracellular pH (pHe) and ROS (reactive oxygen species, ROSe) were in situ monitored by SECM potentiometric mode and SECM-MPSW (multipotential step waveform), respectively. When the signaling cells, including MCF-7, HeLa, and HFF cells, were electrically stimulated, the ROS release of the signal-receiving cells was promoted. By detecting the pH at the cell surface, it was found that more H+ generated by the signaling cells and two cell layers at a shorter distance could both cause the signal-receiving cells to release more ROS, revealing that H+ is one of the signaling molecules of intercellular communication. This SECM-based in situ monitoring strategy provides an effective way to investigate intercellular signal transduction and explore the corresponding mechanism.

Automated summary

A Transwell chamber-based two-layer device combined with scanning electrochemical microscopy (SECM) technology was used for in situ investigation of intercellular signal transduction. The cells in the device were cultured on two layers, with the lower layer for signaling cells and the upper layer for signal-receiving cells. By monitoring the extracellular pH (pHe) and ROS release with SECM, it was found that H+ generated by the signaling cells and a shorter distance between the two cell layers both promoted the release of ROS in the signal-receiving cells, indicating H+ as a signaling molecule of intercellular communication. This SECM-based in situ monitoring strategy provides an effective method to study intercellular signal transduction and explore the underlying mechanism.