In Vivo Monitoring of pH in Subacute PD Mouse Brains with a Ratiometric Electrochemical Microsensor Based on Poly(melamine) Films

In vivo monitoring of cerebral pH is of great significance because its disturbance is related to some pathological processes such as neurodegenerative diseases, for example, Parkinson’s disease (PD). In this study, we developed an electrochemical microsensor based on poly­(melamine) (PMel) films for ratiometric monitoring of pH in subacute PD mouse brains. In this microsensor, PMel films were prepared from a simple electropolymerization approach in a melamine-containing solution, serving as the selective pH recognition membrane undergoing a 2H+/2e– process. Meanwhile, electrochemically oxidized graphene oxide (EOGO) produced a built-in correction signal which helped avoid the environmental interference of the complicated brain systems. The potential difference between the peaks generated from EOGO and PMel gradually decreased with the aqueous pH increasing from 4.0 to 9.0, constituting the detection foundation of the ratiometric electrochemical microsensor (REM). The in vitro studies demonstrated that this proposed method exhibited a high sensitivity (a Nernstian response of −61.35 mV/pH) and remarkable selectivity against amino acids, anions, cations, and biochemical and reactive oxygen species coexisting in the brain. Coupled with its excellent stability and reproducibility and good antibiofouling based on short-term detection, the developed REM could serve as a disposable sensor for the determination of cerebral pH in vivo. Its following successful application in the real-time measurement of pH in the striatum, hippocampus, and cortex of rat brains in the events of global cerebral ischemia/reperfusion verified the reliability of this method. Finally, we adopted this robust REM to systematically analyze and compare the average pH in different regions of normal and subacute PD mouse brains.