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1. An integrated microfluidic chip for studying the effects of neurotransmitters on neurospheroids

   https://doi.org/10.1039/D2LC00755J  

   Mao, Subin; Fonder, Catherine; Rubby, Md Fazlay; Phillips, Gregory J.; Sakaguchi, Donald S.; Que, Long (March 2023, Lab on a Chip)

   To improve our understanding of how the central nervous system functions in health and disease, we report the development of an integrated chip for studying the effects of the neurotransmitters dopamine and serotonin on adult rat hippocampal progenitor cell (AHPC) neurospheroids. This chip allows dopamine or serotonin located in one chamber to diffuse to AHPC neurospheroids cultured in an adjacent chamber through a built-in diffusion barrier created by an array of intentionally misaligned micropillars. The gaps among the micropillars are filled with porous poly(ethylene glycol) (PEG) gel to tune the permeability of the diffusion barrier. An electrochemical sensor is also integrated within the chamber where the neurospheroids can be cultured, thereby allowing monitoring of the concentrations of dopamine or serotonin. Experiments show that concentrations of the neurotransmitters inside the neurospheroid chamber can be increased over a period of several hours to over 10 days by controlling the compositions of the PEG gel inside the diffusion barrier. The AHPC neurospheroids cultured in the chip remain highly viable following dopamine or serotonin treatment. Cell proliferation and neuronal differentiation have also been observed following treatment, revealing that the AHPC neurospheroids are a valuable in vitro brain model for neurogenesis research. Finally, we show that by tuning the permeability of diffusion barrier, we can block transfer of Escherichia coli cells across the diffusion barrier, while allowing dopamine or serotonin to pass through. These results suggest the feasibility of using the chip to better understand the interactions between microbiota and brain via the gut–brain axis. 

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2. Polymer modified carbon fiber-microelectrodes and waveform modifications enhance neurotransmitter metabolite detection

   https://doi.org/10.1039/C8AY02737D  

   Raju, Dilpreet; Mendoza, Alexander; Wonnenberg, Pauline; Mohanaraj, Sanuja; Sarbanes, Mulugeta; Truong, Carly; Zestos, Alexander G. (March 2019, Analytical Methods)

   Carbon-fiber microelectrodes (CFMEs) have been used for several years for the detection of neurotransmitters such as dopamine. Dopamine is a fundamentally important neurotransmitter and is also metabolized at a subsecond timescale. Recently, several metabolites of dopamine have been shown to be physiologically important such as 3-methoxytyramine (3-MT), 3,4-dihydroxyphenylacetic acid (DOPAC), and homovanillic acid (HVA). Many of these neurotransmitter metabolites are currently only detected with microdialysis coupled with liquid chromatography with relatively low temporal and spatial resolution. Current electrochemical methods such as the dopamine waveform (scanning from −0.4 to 1.3 V at 400 V s −1 ) are utilized to electrostatically repel anions such as DOPAC and promote dopamine adsorption to the surface of the electrode. Moreover, polymer coatings such as Nafion have been shown to electrostatically repel anions such as 5-hydroxyindoleacetic acid (5-HIAA). In this study, we develop novel polymer and waveform modifications for enhanced DOPAC detection. Applying the DOPAC waveform (scanning from 0 to 1.3 V at 400 V s −1 ) enhances DOPAC detection significantly because it does not include the negative holding potential of the dopamine waveform. Moreover, positively charged cationic polymers such as polyethyleneimine (PEI) allow for the preconcentration of DOPAC to the surface of the carbon fiber through an electrostatic attraction. The limit of detection for DOPAC for PEI coated CFMEs with the DOPAC waveform applied is 58.2 ± 2 nM as opposed to 291 ± 10 nM for unmodified electrodes applying the dopamine waveform ( n = 4). This work offers promise for the development of novel electrode materials and waveforms for the specific detection of several important biomolecules such as dopamine metabolite neurotransmitters. 

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3. Carbon Nanotube Yarn Microelectrodes Promote High Temporal Measurements of Serotonin Using Fast Scan Cyclic Voltammetry

   https://doi.org/10.3390/s20041173  

   Mendoza, Alexander; Asrat, Thomas; Liu, Favian; Wonnenberg, Pauline; Zestos, Alexander G. (February 2020, Sensors)

   Carbon fiber-microelectrodes (CFMEs) have been the standard for neurotransmitter detection for over forty years. However, in recent years, there have been many advances of utilizing alternative nanomaterials for neurotransmitter detection with fast scan cyclic voltammetry (FSCV). Recently, carbon nanotube (CNT) yarns have been developed as the working electrode materials for neurotransmitter sensing capabilities with fast scan cyclic voltammetry. Carbon nanotubes are ideal for neurotransmitter detection because they have higher aspect ratios enabling monoamine adsorption and lower limits of detection, faster electron transfer kinetics, and a resistance to surface fouling. Several methods to modify CFMEs with CNTs have resulted in increases in sensitivity, but have also increased noise and led to irreproducible results. In this study, we utilize commercially available CNT-yarns to make microelectrodes as enhanced neurotransmitter sensors for neurotransmitters such as serotonin. CNT-yarn microelectrodes have significantly higher sensitivities (peak oxidative currents of the cyclic voltammograms) than CFMEs and faster electron transfer kinetics as measured by peak separation (ΔEP) values. Moreover, both serotonin and dopamine are adsorption controlled to the surface of the electrode as measured by scan rate and concentration experiments. CNT yarn microelectrodes also resisted surface fouling of serotonin onto the surface of the electrode over thirty minutes and had a wave application frequency independent response to sensitivity at the surface of the electrode. 

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4. Continuous Real-Time Detection of Serotonin Using an Aptamer-Based Electrochemical Biosensor

   https://doi.org/10.3390/bios13110983  

   Ahmad, Habib_M N; Andrade, Arturo; Song, Edward (November 2023, Biosensors)

   Serotonin (5-HT) is a critical neurotransmitter involved in many neuronal functions, and 5-HT depletion has been linked to several mental diseases. The fast release and clearance of serotonin in the extracellular space, low analyte concentrations, and a multitude of interfering species make the detection of serotonin challenging. This work presents an electrochemical aptamer-based biosensing platform that can monitor 5-HT continuously with high sensitivity and selectivity. Our electrochemical sensor showed a response time of approximately 1 min to a step change in the serotonin concentration in continuous monitoring using a single-frequency EIS (electrochemical impedance spectroscopy) technique. The developed sensing platform was able to detect 5-HT in the range of 25–150 nM in the continuous sample fluid flow with a detection limit (LOD) of 5.6 nM. The electrochemical sensor showed promising selectivity against other species with similar chemical structures and redox potentials, including dopamine (DA), norepinephrine (NE), L-tryptophan (L-TP), 5-hydroxyindoleacetic acid (5-HIAA), and 5-hydroxytryptophan (5-HTP). The proposed sensing platform is able to achieve high selectivity in the nanomolar range continuously in real-time, demonstrating the potential for monitoring serotonin from neurons in organ-on-a-chip or brain-on-a-chip-based platforms. 

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5. Drosophila mutants lacking the glial neurotransmitter-modifying enzyme Ebony exhibit low neurotransmitter levels and altered behavior

   https://doi.org/10.1038/s41598-023-36558-7  

   Pantalia, Meghan; Lin, Zhi; Tener, Samantha J.; Qiao, Bing; Tang, Grace; Ulgherait, Matthew; O’Connor, Reed; Delventhal, Rebecca; Volpi, Julia; Syed, Sheyum; et al (June 2023, Scientific Reports)

   Abstract Inhibitors of enzymes that inactivate amine neurotransmitters (dopamine, serotonin), such as catechol-O-methyltransferase (COMT) and monoamine oxidase (MAO), are thought to increase neurotransmitter levels and are widely used to treat Parkinson's disease and psychiatric disorders, yet the role of these enzymes in regulating behavior remains unclear. Here, we investigated the genetic loss of a similar enzyme in the model organismDrosophila melanogaster. Because the enzyme Ebony modifies and inactivates amine neurotransmitters, its loss is assumed to increase neurotransmitter levels, increasing behaviors such as aggression and courtship and decreasing sleep. Indeed,ebonymutants have been described since 1960 as aggressive mutants, though this behavior has not been quantified. Using automated machine learning-based analyses, we quantitatively confirmed thatebonymutants exhibited increased aggressive behaviors such as boxing but also decreased courtship behaviors and increased sleep. Through tissue-specific knockdown, we found thatebony’s role in these behaviors was specific to glia. Unexpectedly, direct measurement of amine neurotransmitters inebonybrains revealed that their levels were not increased but reduced. Thus, increased aggression is the anomalous behavior for this neurotransmitter profile. We further found thatebonymutants exhibited increased aggression only when fighting each other, not when fighting wild-type controls. Moreover, fights betweenebonymutants were less likely to end with a clear winner than fights between controls or fights betweenebonymutants and controls. Inebonyvs. control fights,ebonymutants were more likely to win. Together, these results suggest thatebonymutants exhibit prolonged aggressive behavior only in a specific context, with an equally dominant opponent. 

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