Optimized Fabrication of Carbon-Fiber Microbiosensors for Codetection of Glucose and Dopamine in Brain Tissue.

Dopamine (DA) signaling is critically important in striatal function, and this metabolically demanding process is fueled largely by glucose. However, DA and glucose are typically studied independently and, as such, the precise relationship between DA release and glucose availability remains unclear. Fast-scan cyclic voltammetry (FSCV) is commonly coupled with carbon-fiber microelectrodes to study DA transients. These microelectrodes can be modified with glucose oxidase (GOx) to generate microbiosensors capable of simultaneously quantifying real-time and physiologically relevant fluctuations of glucose, a nonelectrochemically active substrate, and DA, which is readily oxidized and reduced at the electrode surface. A chitosan hydrogel can be electrodeposited to entrap the oxidase enzyme on the sensor surface for stable, sensitive, and selective codetection of glucose and DA using FSCV. This strategy can also be used to entrap lactate oxidase on the carbon-fiber surface for codetection of lactate and DA. However, these custom probes are individually fabricated by hand, and performance is variable. This study characterizes the physical nature of the hydrogel and its effects on the acquired electrochemical data in the detection of glucose (2.6 mM) and DA (1 µM). The results demonstrate that the electrodeposition of the hydrogel membrane is improved using a linear potential sweep rather than a direct step to the target potential. Electrochemical impedance spectroscopy data relate information on the physical nature of the electrode/solution interface to the electrochemical performance of bare and enzyme-modified carbon-fiber microelectrodes. The electrodeposition waveform and scan rate were characterized for optimal membrane formation and performance. Finally, codetection of both DA/glucose and DA/lactate was demonstrated in intact rat striatum using probes fabricated according to the optimized protocol. Overall, this work improves the reliable fabrication of carbon-fiber microbiosensors for codetection of DA and important energetic substrates that are locally delivered to the recording site to meet metabolic demand.

Simultaneous, Real-Time Detection of Glutamate and Dopamine in Rat Striatum Using Fast-Scan Cyclic Voltammetry.

Glutamate and dopamine (DA) represent two key contributors to striatal functioning, a region of the brain that is essential to motor coordination and motivated behavior. While electroanalytical techniques can be utilized for rapid, spatially resolved detection of DA in the interferent-rich brain environment, glutamate, a nonelectroactive analyte, cannot be directly detected using electroanalytical techniques. However, it can be probed using enzyme-based sensors, which generate an electroactive reporter in the presence of glutamate. The vast majority of glutamate biosensors have relied on amperometric sensing, which is an inherently nonselective detection technique. This approach necessitates the use of complex and performance-limiting modifications to ensure the desired single-analyte specificity. Here, we present a novel glutamate microbiosensor fabricated on a carbon-fiber microelectrode substrate and coupled with fast-scan cyclic voltammetry (FSCV) to enable the simultaneous quantification of glutamate and DA at single recording sites in the brain, which is impossible when using typical amperometric approaches. The glutamate microbiosensors were characterized for sensitivity, stability, and selectivity by using a voltammetric waveform optimized for the simultaneous detection of both species. The applicability of these sensors for the investigation of neural circuits was validated in the rat ventral striatum. Electrically evoked glutamate and DA release were recorded at single-micrometer-scale locations before and after pharmacological manipulation of glutamatergic signaling. Our novel glutamate microbiosensor advances the state of the art by providing a powerful tool for probing coordination between these two species in a way that has previously not been possible.

A Student Perspective on the 18th Monitoring Molecules in Neuroscience Meeting in Lyon.

After being postponed twice due to the global COVID-19 pandemic, approximately 200 scientists gathered in Lyon, France, in late June 2022 for the 18th Biennial Monitoring Molecules in Neuroscience (MMiN) Research Conference. Although there were unprecedented challenges involved with coordinating the 18th MMiN conference, the meeting was a huge success. The meeting provided a wonderful opportunity for young neuroscientists to network and learn about the current state of molecular monitoring in neuroscience research. The topics spanned advancements in well-established analytical techniques to novel method development. Some of the noteworthy techniques expediting our understanding of circuit-level neurochemical function include multiplexed detection of numerous neurochemicals, well-established sensors leveraging enzymes and other biologic components, and the development of diverse, customizable genetically encoded sensors.

Simultaneous voltammetric detection of glucose and lactate fluctuations in rat striatum evoked by electrical stimulation of the midbrain.

Glucose and lactate provide energy for cellular function in the brain and serve as an important carbon source in the synthesis of a variety of biomolecules. Thus, there is a critical need to quantitatively monitor these molecules in situ on a time scale commensurate with neuronal function. In this work, carbon-fiber microbiosensors were coupled with fast-scan cyclic voltammetry to monitor glucose and lactate fluctuations at a discrete site within rat striatum upon electrical stimulation of the midbrain projection to the region. Systematic variation of stimulation parameters revealed the distinct dynamics by which glucose and lactate responded to the metabolic demand of synaptic function. Immediately upon stimulation, extracellular glucose and lactate availability rapidly increased. If stimulation was sufficiently intense, concentrations then immediately fell below baseline in response to incurred metabolic demand. The dynamics were dependent on stimulation frequency, such that more robust fluctuations were observed when the same number of pulses was delivered at a higher frequency. The rates at which glucose was supplied to, and depleted from, the local recording region were dependent on stimulation intensity, and glucose dynamics led those of lactate in response to the most substantial stimulations. Glucose fluctuated over a larger concentration range than lactate as stimulation duration increased, and glucose fell further from baseline concentrations. These real-time measurements provide an unprecedented direct comparison of glucose and lactate dynamics in response to metabolic demand elicited by neuronal activation. Graphical abstract.