Single-Vesicle Electrochemistry in Fresh Brain Slices Enables In Situ Quantification of Vesicular Monoamine

The quantitative analysis of vesicular neurotransmitters in neurons in situ is paramount for investigating neurodegenerative disorders, such as Alzheimer's disease and Parkinson's disease (PD). Unfortunately, a direct approach for monitoring neurotransmitter chemistry in single vesicles in fresh brain tissue has remained inaccessible so far. Here, we introduce an innovative platform of single-vesicle electrochemistry (SVE) in fresh brain tissue, enabling the quantification of neurotransmitters at the single-vesicle level for both soma and varicosity. Utilizing this methodology, we investigated a PD animal model, which demonstrated a significant reduction in both vesicular dopamine (DA) storage and exocytotic release in DA neurons in the substantia nigra. Furthermore, SVE unveiled the heterogeneous nature of chemical neurotransmission among DA neurons across different brain regions. Importantly, this tissue-based SVE approach can be extended to quantify other monoamine neurotransmitters, such as norepinephrine and serotonin, at the single-vesicle level. The introduction of this methodology marks a significant advancement, offering a novel avenue to explore neurological and psychiatric disorders through the lens of neurotransmitter signaling in the mammalian brain. This breakthrough has the potential to deepen our understanding of the underlying mechanisms of these complex disorders and may pave the way for the development of targeted therapeutic interventions.