We constructed a composite membrane-modified carbon fiber microelectrode for ascorbic acid detection and used it to observe the process of glutamate-induced brain cytotoxic edema with AA release and reveal the mechanism. Our study is significant for monitoring neurochemicals, understanding the molecular basis of nervous system diseases, and discovering biomarkers of brain diseases.
Although neuroelectrochemical sensing technology offers unique benefits for neuroscience research, its application is limited by substantial interference in complex brain environments while ensuring biosafety requirements. In this study, we introduced poly(3-hexylthiophene) (P3HT) and nitrogen-doped multiwalled carbon nanotubes (N-MWCNTs) to construct a composite membrane-modified carbon fiber microelectrode (CFME/P3HT-N-MWCNTs) for ascorbic acid (AA) detection. The microelectrode presented good linearity, selectivity, stability, antifouling, and biocompatibility and exhibited great performance for application in neuroelectrochemical sensing. Subsequently, we applied CFME/P3HT-N-MWCNTs to monitor AA release from in vitro nerve cells, ex vivo brain slices, and in vivo living rat brains and determined that glutamate can induce cell edema and AA release. We also found that glutamate activated the N-methyl-d-aspartic acid receptor, which enhanced Na+ and Cl- inflow to induce osmotic stress, resulting in cytotoxic edema and ultimately AA release. This study is the first to observe the process of glutamate-induced brain cytotoxic edema with AA release and to reveal the mechanism. Our work can benefit the application of P3HT in in vivo implant microelectrode construction to monitor neurochemicals, understand the molecular basis of nervous system diseases, and discover certain biomarkers of brain diseases.
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