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Fast scan cyclic voltammetry (FSCV) at carbon fiber microelectrodes (CFMs) is a method traditionally used for real-time quantification of neurotransmitters in biological systems. Reliable calibration of CFMs is essential for converting FSCV signals to analyte concentrations and generally employs flow injection analysis (FIA) performed with flow cells fabricated in-house. Such FSCV FIA cells often require significant and ongoing troubleshooting with pulsing, leaking, flow inconsistencies and dead volume being major causes of common challenges. In this work, we address these issues by creating a robust, plug-and-play FSCV flow cell. This novel design permits reproducible, high-precision, and stable flow injection profiles using low-cost materials to improve FSCV calibration. The ready-to-print computer-aided designs and hardware list are provided.

Depression pathology remains elusive. The monoamine hypothesis has placed much focus on serotonin, but due to the variable clinical efficacy of monoamine reuptake inhibitors, the community is looking for alternative therapies such as ketamine (synaptic plasticity and neurogenesis theory of antidepressant action). There is evidence that different classes of antidepressants may affect serotonin levels; a notion we test here. We measure hippocampal serotonin in mice with voltammetry and study the effects of acute challenges of antidepressants. We find that pseudo-equivalent doses of these drugs similarly raise ambient serotonin levels, despite their differing pharmacodynamics because of differences in Uptake 1 and 2, rapid SERT trafficking and modulation of serotonin by histamine. These antidepressants have different pharmacodynamics but have strikingly similar effects on extracellular serotonin. Our findings suggest that serotonin is a common thread that links clinically effective antidepressants, synergizing different theories of depression (synaptic plasticity, neurogenesis and the monoamine hypothesis).

Psychiatric diseases, like depression, largely affect the central nervous system (CNS). While the underlying neuropathology of depressive illness remains to be elucidated, several hypotheses have been proposed as molecular underpinnings for major depressive disorder, including the monoamine hypothesis and the cytokine hypothesis. The monoamine hypothesis has been largely supported by the pharmaceuticals that target monoamine neurotransmitters as a treatment for depression. However, these antidepressants have come under scrutiny due to their limited clinical efficacy, side effects, and delayed onset of action. The more recent, cytokine hypothesis of depression is supported by the ability of immune-active agents to induce "sickness behaviour" akin to that seen with depression. However, treatments that more selectively target inflammation have yielded inconsistent antidepressive results. As such, neither of these hypotheses can fully explain depressive illness pathology, implying that the underlying neuropathological mechanisms may encompass aspects of both theories. The goal of the current review is to integrate these two well-studied hypotheses and to propose a role for histamine as a potential unifying factor that links monoamines to cytokines. Additionally, we will focus on stress-induced depression, to provide an updated perspective of depressive illness research and thereby identify new potential targets for the treatment of major depressive disorder.

Deep brain stimulation (DBS) is an invasive neuromodulatory technique used for the treatment of individuals with neurological disorders. Open-loop DBS delivers fixed stimulation pulses while closed-loop DBS senses the state of disorder and delivers stimulation pulses optimized for the sensed state adaptively. This review focuses on current miniature closed-loop DBS devices. The latest progress in stimulation techniques and electrode configurations are discussed. We cover common circuit configurations used for stimulation and outline the latest methods for delivering rectangular and non-rectangular stimulation waveforms. Recent advances in feedback control for closing the loop in DBS are highlighted, as are cutting edge developments in artificial intelligence (AI) methods that analyse the sensed data to determine the state of disorder. Finally, we critically discuss present issues and future directions of miniaturized closed-loop DBS.

Carbon is the material of choice for electroanalysis of biological systems, being particularly applicable to neurotransmitter analysis as carbon fiber microelectrodes (CFMs). CFMs are most often applied to dopamine detection; however, the scope of CFM analysis has rapidly expanded over the last decade with our laboratory's focus being on improving serotonin detection at CFMs, which we achieved in the past via Nafion modification. We began this present work by seeking to optimize this modification to gain increased analytical sensitivity toward serotonin under the assumption that exposure of bare carbon to the in vivo environment rapidly deteriorates analytical performance. However, we were unable to experimentally verify this assumption and found that electrodes that had been exposed to the in vivo environment were more sensitive to evoked and ambient dopamine. We hypothesized that high in vivo concentrations of ambient extracellular glutamate could polymerize with a negative charge onto CFMs and facilitate response to dopamine. We verified this polymerization electrochemically and characterized the mechanisms of deposition with micro- and nano-imaging. Importantly, we identified that the application of 1.3 V as a positive upper waveform limit is a crucial factor for facilitating glutamate polymerization, thus improving analytical performance. Critically, information gained from these dopamine studies were extended to an in vivo environment where a 2-fold increase in sensitivity to evoked serotonin was achieved. Thus, we present here the novel finding that innate aspects of the in vivo environment are auspicious for detection of dopamine and serotonin at carbon fibers, offering a solution to our goal of an improved fast-scan cyclic voltammetry serotonin detection paradigm.

Commonly prescribed selective serotonin reuptake inhibitors (SSRIs) inhibit the serotonin transporter to correct a presumed deficit in extracellular serotonin signaling during depression. These agents bring clinical relief to many who take them; however, a significant and growing number of individuals are resistant to SSRIs. There is emerging evidence that inflammation plays a significant role in the clinical variability of SSRIs, though how SSRIs and inflammation intersect with synaptic serotonin modulation remains unknown. In this work, we use fast in vivo serotonin measurement tools to investigate the nexus between serotonin, inflammation, and SSRIs. Upon acute systemic lipopolysaccharide (LPS) administration in male and female mice, we find robust decreases in extracellular serotonin in the mouse hippocampus. We show that these decreased serotonin levels are supported by increased histamine activity (because of inflammation), acting on inhibitory histamine H3 heteroreceptors on serotonin terminals. Importantly, under LPS-induced histamine increase, the ability of escitalopram to augment extracellular serotonin is impaired because of an off-target action of escitalopram to inhibit histamine reuptake. Finally, we show that a functional decrease in histamine synthesis boosts the ability of escitalopram to increase extracellular serotonin levels following LPS. This work reveals a profound effect of inflammation on brain chemistry, specifically the rapidity of inflammation-induced decreased extracellular serotonin, and points the spotlight at a potentially critical player in the pathology of depression, histamine. The serotonin/histamine homeostasis thus, may be a crucial new avenue in improving serotonin-based treatments for depression. SIGNIFICANCE STATEMENT Acute LPS-induced inflammation (1) increases CNS histamine, (2) decreases CNS serotonin (via inhibitory histamine receptors), and (3) prevents a selective serotonin reuptake inhibitor (SSRI) from effectively increasing extracellular serotonin. A targeted depletion of histamine recovers SSRI-induced increases in extracellular hippocampal serotonin.

Fast-scan cyclic voltammetry at carbon fiber microelectrodes measures low concentrations of analytes in biological systems. There are ongoing efforts to simplify FSCV analysis and several custom platforms are available for filtering and multi-modal analysis of FSCV signals but there is no single, easily accessible platform that has capacity for all these features. Here we present The Analysis Kid: a free, open-source cloud application that does not require a specialized runtime environment and is easily accessible via common browsers. We show that a user-friendly interface can analyze multi-platform file formats to provide multimodal visualization of FSCV color plots with digital background subtraction. We highlight key features that allow interactive calibration and parametric analysis via peak finding algorithms to automatically detect the maximum amplitude, area under the curve and clearance rate of the signal. Finally, The Analysis Kid enables semi-automatic fitting of data with Michaelis Menten kinetics with single or dual reuptake models. The Analysis Kid can be freely accessed at https://analysis-kid.herokuapp.com/. The web application code is found, under an MIT license, at https://github.com/sermeor/The-Analysis-Kid.

The coauthors have been working together for ten years on serotonin, dopamine, and histamine and their connection to neuropsychiatric illnesses. Hashemi has pioneered many new experimental techniques for measuring serotonin and histamine in real time in the extracellular space in the brain. Best, Reed, and Nijhout have been making mathematical models of brain metabolism to help them interpret Hashemi’s data. Hashemi demonstrated that brain histamine inhibits serotonin release, giving a direct mechanism by which inflammation can cause a decrease in brain serotonin and therefore depression. Many new biological phenomena have come out of their joint research including 1) there are two different reuptake mechanisms for serotonin; 2) the effect of the serotonin autoreceptors is not instantaneous and is long-lasting even when the extracellular concentrations have returned to normal; 3) that mathematical models of serotonin metabolism and histamine metabolism can explain Hashemi’s experimental data; 4) that variation in serotonin autoreceptors may be one of the causes of serotonin-linked mood disorders. Here we review our work in recent years for biological audiences, medical audiences, and researchers who work on mathematical modeling of biological problems. We discuss the experimental techniques, the creation and investigation of mathematical models, and the consequences for neuropsychiatric diseases.

Depression is quickly becoming one of the world's most pressing public health crises, and there is an urgent need for better diagnostics and therapeutics. Behavioral models in animals and humans have not adequately addressed the diagnosis and treatment of depression, and biomarkers of mental illnesses remain ill-defined. It has been very difficult to identify biomarkers of depression because of

Serotonin's involvement in many physiological processes including anxiety, stress, aggression, and mood has been known for decades; however, a more in-depth characterization of serotonin function in the brain is still missing. Insufficient progress in our understanding of serotonin function is in part due to the lack of effective tools to selectively measure this neurotransmitter on a relevant time scale in vivo. An analytical technique for serotonin measurements, fast-scan cyclic voltammetry was recently pioneered in anesthetized rodents and is beginning to shed light on the complexity of serotonergic activity on a subsecond time scale. Complementary, due to even more recent technological developments, ambient serotonin levels can now also be assessed with high temporal resolution using a method called fast-scan controlled-adsorption voltammetry. Here, we discuss the traditional experimental challenges of measuring serotonin, how voltammetry overcomes some of these difficulties, and future plans to better understand the function of serotonin in behavior.

Histamine is well known for mediating peripheral inflammation, but histamine is also found in high concentrations in the brain where amongst other roles, this amine is thought to be neuromodulator. Neuromodulation is critical for brain function, yet histamine dynamics are very difficult to measure and thus several fundamental aspects of the mechanisms that control the extracellular and modulatory behavior of this messenger remain undefined. In this work we undertake the first in-depth characterization of in vivo histamine dynamics in real time using fast-scan cyclic voltammetry at carbon fiber microelectrodes. We measure electrically evoked histamine in the mouse hypothalamus and find that histamine release is sensitive to pharmacological manipulation at the level of synthesis, packaging, autoreceptor control of release, and metabolism. We find two breakthrough aspects of histamine modulation. First, there are differences in H3 receptor regulation of histamine between sexes showing that histamine release in female mice is more much tightly regulated than in male mice under H3 or inflammatory drug challenge. We hypothesize that this finding may contribute to hormone-mediated neuroprotection mechanisms in female mice. Second, we find that a high dose of a commonly available antihistamine, the H1 receptor inverse agonist diphenhydramine, rapidly decreases serotonin levels. This high dose is considered overdose; however, this finding highlights the sheer significance of better consideration of the modulatory nuances of histamine on serotonin.We, thus, present the first in depth in vivo characterization of fast histamine dynamics and highlight two breakthrough regulatory aspects of this elusive modulator. The implications of our study are new avenues to better understand and treat histamine related disorders of the brain (such as neuroinflammation), emphasizing that sex and modulation (of serotonin) are critical factors to consider when studying/designing new histamine targeting therapeutics.

Histamine is well known for mediating peripheral inflammation, but histamine is also found in high concentrations in the brain where amongst other roles, this amine is thought to be neuromodulator. Neuromodulation is critical for brain function, yet histamine dynamics are very difficult to measure and thus several fundamental aspects of the mechanisms that control the extracellular and modulatory behavior of this messenger remain undefined. In this work we undertake the first in-depth characterization of in vivo histamine dynamics in real time using fast-scan cyclic voltammetry at carbon fiber microelectrodes. We measure electrically evoked histamine in the mouse hypothalamus and find that histamine release is sensitive to pharmacological manipulation at the level of synthesis, packaging, autoreceptor control of release, and metabolism. We find two breakthrough aspects of histamine modulation. First, there are differences in H3 receptor regulation of histamine between sexes showing that histamine release in female mice is more much tightly regulated than in male mice under H3 or inflammatory drug challenge. We hypothesize that this finding may contribute to hormone-mediated neuroprotection mechanisms in female mice. Second, we find that a high dose of a commonly available antihistamine, the H1 receptor inverse agonist diphenhydramine, rapidly decreases serotonin levels. This high dose is considered overdose; however, this finding highlights the sheer significance of better consideration of the modulatory nuances of histamine on serotonin.We, thus, present the first in depth in vivo characterization of fast histamine dynamics and highlight two breakthrough regulatory aspects of this elusive modulator. The implications of our study are new avenues to better understand and treat histamine related disorders of the brain (such as neuroinflammation), emphasizing that sex and modulation (of serotonin) are critical factors to consider when studying/designing new histamine targeting therapeutics.

Fast-scan adsorption-controlled voltammetry (FSCAV) was recently derived from fast-scan cyclic voltammetry to estimate the absolute concentrations of neurotransmitters by using the innate adsorption properties of carbon fiber microelectrodes. This technique has improved our knowledge of serotonin dynamics

While the neurochemistry that underpins the behavioral phenotypes of depression is the subject of many studies, oxidative stress caused by the inflammation comorbid with depression has not adequately been addressed. In this study, we described novel antidepressant-antioxidant agents consisting of selenium-modified fluoxetine derivatives to simultaneously target serotonin reuptake (antidepressant action) and oxidative stress. Excitingly, we show that one of these agents (

Carbon is the material of choice for electroanalysis of biological systems, being particularly applicable to neurotransmitter analysis as carbon fiber microelectrodes (CFMs). CFMs are most often applied to dopamine detection; however, the scope of CFM analysis has rapidly expanded over the last decade with our laboratory's focus being on improving serotonin detection at CFMs, which we achieved in the past

Neuromodulators are critical regulators of the brain's signaling processes, and thus they are popular pharmacological targets for psychoactive therapies. It is clear that monoamine uptake mechanisms are complicated and subject to multiple uptake mechanisms. Uptake 1 describes uptake of the monoamine via its designated transporter (SERT for serotonin, NET for norepinephrine, and DAT for dopamine), whereas Uptake 2 details multiple transporter types on neurons and glia taking up different types of modulators, not necessarily specific to the monoamine. While Uptake 1 processes have been well-studied over the past few decades, Uptake 2 mechanisms have remained more difficult to study because of the limitations in methods that have the sensitivity and spatiotemporal resolution to look at the subtleties in uptake profiles. In this chapter we review the different experimental approaches that have yielded important information about Uptake 2 mechanisms in vivo. The techniques (scintillation microspectrophotometry, microdialysis, chronoamperometry, and voltammetry) are described in detail, and pivotal studies associated with each method are highlighted. It is clear from these reviewed works that Uptake 2 processes are critical to consider to advance our understanding of the brain and develop effective neuropsychiatric therapies.

It is important to monitor serotonin neurochemistry in the context of brain disorders. Specifically, a better understanding of biophysical alterations and associated biochemical functionality within subregions of the brain will enable better of understanding of diseases such as depression. Fast voltammetric tools at carbon fiber microelectrodes provide an opportunity to make direct evoked and ambient serotonin measurements in vivo in mice. In this study, we characterize novel stimulation and measurement circuitries for serotonin analyses in brain regions relevant to psychiatric disease. Evoked and ambient serotonin in these brain areas, the CA2 region of the hippocampus and the medial prefrontal cortex, are compared to ambient and evoked serotonin in the substantia nigra pars reticulata, an area well established previously for serotonin measurements with fast voltammetry. Stimulation of a common axonal location evoked serotonin in all three brain regions. Differences are observed in the serotonin release and reuptake profiles between these three brain areas which we hypothesize to arise from tissue physiology heterogeneity around the carbon fiber microelectrodes. We validate this hypothesis mathematically and via confocal imaging. We thereby show that fast voltammetric methods can provide accurate information about local physiology and highlight implications for chemical mapping. Cover Image for this issue: doi: 10.1111/jnc.14739.

Between 30% and 60% of HIV seropositive individuals develop symptoms of clinical depression and/or apathy. Dopamine and serotonin are associated with motivational alterations; however, histamine is less well studied. In the present study, we used fast-scan cyclic voltammetry in HIV-1 transgenic (Tg) rats to simultaneously analyze the kinetics of nucleus accumbens dopamine (DA), prefrontal cortical serotonin (5-HT), and hypothalamic histamine (HA). For voltammetry, subjects were 15 HIV-1 Tg (7 male, 8 female) and 20 F344/N (11 male, 9 female) adult rats. Both serotonergic and dopaminergic release and reuptake kinetics were decreased in HIV-1 Tg animals relative to controls. In contrast, rates of histamine release and reuptake increased in HIV-1 Tg rats. Additionally, we used immunohistochemical (IHC) methods to identify histaminergic neurons in the tuberomammillary nucleus (TMN) of the hypothalamus. For IHC, subjects were 9 HIV-1 Tg (5 male, 4 female) and 9 F344/N (5 male, 4 female) adult rats. Although the total number of TMN histaminergic cells did not differ between HIV-1 Tg rats and F344/N controls, a significant sex effect was found; with females having an increased number of histaminergic neurons, relative to males. Collectively, these findings illustrate neurochemical alterations that potentially underlie or exacerbate the pathogenesis of clinical depression and/or apathy in HIV-1.

Metal speciation analysis has wide applicability and reveals how and when metals are available to engage in chemical processes. Real systems can be physically hard to reach, chemically dynamic, and sample collection and pretreatment fundamentally alter speciation. Therefore, there is great interest in in situ trace metal speciation sensors that can be integrated into on-site, portable analysis devices. Electrochemistry is an attractive method since the sensing component can be fashioned into a submersible probe. In this review, we define a set of criteria that an electrochemical approach must conform to for in situ metal analysis and review methods that closely follow these criteria. As an emerging technique, fast-scan cyclic voltammetry is introduced. Next, design and material of analysis probes is detailed, followed by a discussion of portable instrumentation. Finally, studies that show promise towards a portable, in situ metal analysis device are showcased and ongoing analytical challenges are outlined.

Fast-scan cyclic voltammetry (FSCV) is an analytical tool used to probe neurochemical processes in real-time. A major drawback for specialized applications of FSCV is that instrumentation must be constructed or modified in-house by those with expertise in electronics. One such specialized application is the newly developed fast-scan controlled-adsorption voltammetry (FSCAV) that measures basal (tonic) in vivo dopamine and serotonin concentrations. FSCAV requires additional software and equipment (an operational amplifier coupled to a transistor-transistor logic) allowing the system to switch between applying a FSCV waveform and a constant potential to the working electrode. Herein we describe a novel, simplified switching component to facilitate the integration of FSCAV into existing FSCV instruments, thereby making this method more accessible to the community. Specifically, we employ two light emitting diodes (LEDs) to generate the voltage needed to drive a NPN bipolar junction transistor, substantially streamlining the circuitry and fabrication of the switching component. We performed in vitro and in vivo analyses to compare the new LED circuit vs. the original switch. Our data shows that the novel simplified switching component performs equally well when compared to traditional instrumentation. Thus, we present a new, simplified scheme to perform FSCAV that is cheap, simple, and easy to construct by individuals without a background in engineering and electronics. Keywords: Dopamine, Fast-scan cyclic voltammetry, Basal, Circuit, Instrumentation, Component

Fast-scan cyclic voltammetry at carbon fiber microelectrodes measures low concentrations of analytes in biological systems. There are ongoing efforts to simplify FSCV analysis and several custom platforms are available for filtering and multi-modal analysis of FSCV signals but there is no single, easily accessible platform that has capacity for all these features. Here we present The Analysis Kid: a free, open-source cloud application that does not require a specialized runtime environment and is easily accessible via common browsers. We show that a user-friendly interface can analyze multi-platform file formats to provide multimodal visualization of FSCV color plots with digital background subtraction. We highlight key features that allow interactive calibration and parametric analysis via peak finding algorithms to automatically detect the maximum amplitude, area under the curve and clearance rate of the signal. Finally, The Analysis Kid enables semi-automatic fitting of data with Michaelis Menten kinetics with single or dual reuptake models. The Analysis Kid can be freely accessed at https://analysis-kid.herokuapp.com/. The web application code is found, under an MIT license, at https://github.com/sermeor/The-Analysis-Kid.

Developing tools that are able to monitor transient neurochemical dynamics is important to decipher brain chemistry and function. Multifunctional polymer-based fibers have been recently applied to monitor and modulate neural activity. Here, we explore the potential of polymer fibers comprising six graphite-doped electrodes and two microfluidic channels within a flexible polycarbonate body as a platform for sensing pH and neurometabolic lactate. Electrodes were made into potentiometric sensors (responsive to pH) or amperometric sensors (lactate biosensors). The growth of an iridium oxide layer made the fiber electrodes responsive to pH in a physiologically relevant range. Lactate biosensors were fabricated via platinum black growth on the fiber electrode, followed by an enzyme layer, making them responsive to lactate concentration. Lactate fiber biosensors detected transient neurometabolic lactate changes in an in vivo mouse model. Lactate concentration changes were associated with spreading depolarizations, known to be detrimental to the injured brain. Induced waves were identified by a signature lactate concentration change profile and measured as having a speed of ∼2.7 mm/min (n = 4 waves). Our work highlights the potential applications of fiber-based biosensors for direct monitoring of brain metabolites in the context of injury.

Neuromodulators are critical regulators of the brain's signaling processes, and thus they are popular pharmacological targets for psychoactive therapies. It is clear that monoamine uptake mechanisms are complicated and subject to multiple uptake mechanisms. Uptake 1 describes uptake of the monoamine via its designated transporter (SERT for serotonin, NET for norepinephrine, and DAT for dopamine), whereas Uptake 2 details multiple transporter types on neurons and glia taking up different types of modulators, not necessarily specific to the monoamine. While Uptake 1 processes have been well-studied over the past few decades, Uptake 2 mechanisms have remained more difficult to study because of the limitations in methods that have the sensitivity and spatiotemporal resolution to look at the subtleties in uptake profiles. In this chapter we review the different experimental approaches that have yielded important information about Uptake 2 mechanisms in vivo. The techniques (scintillation microspectrophotometry, microdialysis, chronoamperometry, and voltammetry) are described in detail, and pivotal studies associated with each method are highlighted. It is clear from these reviewed works that Uptake 2 processes are critical to consider to advance our understanding of the brain and develop effective neuropsychiatric therapies.

Clinical studies indicate that obese individuals have an increased risk of developing co-morbid depressive illness and that these patients have reduced responses to antidepressant therapy, including selective serotonin reuptake inhibitors (SSRIs). Obesity, a condition of chronic mild inflammation including obesity-induced neuroinflammation, is proposed to contribute to decreases in synaptic concentrations of neurotransmitters like serotonin (5HT) by decreasing 5HT synthesis in the dorsal raphe nucleus (DRN) and/or affecting 5HT reuptake in DRN target regions like the hippocampus. In view of these observations, the goal of the current study was to examine inflammatory markers and serotonergic dynamics in co-morbid obesity and depression. Biochemical and behavioral assays revealed that high-fat diet produced an obesity and depressive-like phenotype in one cohort of rats and that these changes were marked by increases in key pro-inflammatory cytokines in the hippocampus. In real time using fast scan cyclic voltammetry (FSCV), we observed no changes in basal levels of hippocampal 5HT; however responses to escitalopram were significantly impaired in the hippocampus of obese rats compared to diet resistant rats and control rats. Further studies revealed that these neurochemical observations could be explained by increases in serotonin transporter (SERT) expression in the hippocampus driven by elevated neuroinflammation. Collectively, these results demonstrate that obesity-induced increases in neuroinflammation adversely affect SERT expression in the hippocampus of obese rats, thereby providing a potential synaptic mechanism for reduced SSRI responsiveness in obese subjects with co-morbid depressive illness.

Jean-François Masson, Parastoo Hashemi and Martyn Boutelle introduce the Analyst themed collection on Analytical Science in Neurochemistry.

There is great interest in rapidly monitoring metals of biological and environmental interest. Electrochemistry is traditionally a powerful tool for metal analysis but can be limited by its scope and low temporal resolution. The scope is limited by the potential window of the working electrode and rapid analysis is limited, in part, by the need for nucleation/growth for preconcentration. In prior work, we showed that a rapid equilibrium mediated preconcentration process facilitated fast scan cyclic voltammetry (FSCV) responses to Cu(II) and Pb(II) at carbon fiber microelectrodes (CFMs). In this manuscript, we apply this same principle to Ca(II), Al(III), Mg(II), and Zn(II), metal ions that are traditionally difficult to electroanalyze. We demonstrate FSCV reduction peaks for these four metals whose positions and amplitudes are dependent on scan rate. The adsorption profiles of these ions onto CFMs follow Langmuir's theory for monolayer coverage. We calculate the thermodynamic equilibrium constant of metal adsorption onto CFMs and find that these constants follow the same order as those previously reported by other groups on other activated carbon materials. Finally, a real-time complexation study is performed with ligands that have preference for divalent or multivalent ions to probe the selectivity of the real-time signal. We observe a linear relationship between formation constant ( k

Background Histamine (HA), a small molecule that is synthesized from the amino acid histidine, plays an important role in the immune system where it is associated with allergies, inflammation, and T-cell regulation. In the brain, histamine is stored in mast cells and other non-neuronal cells and also acts as a neurotransmitter. The histamine neuron cell bodies are in the tuberomammillary (TM) nucleus of the hypothalamus and these neurons send projections throughout the central nervous system (CNS), in particular to the cerebral cortex, amygdala, basal ganglia, hippocampus, thalamus, retina, and spinal cord. HA neurons make few synapses, but release HA from the cell bodies and from varicosities when the neurons fire. Thus the HA neural system seems to modulate and control the HA concentration in projection regions. It is known that high HA levels in the extracellular space inhibit serotonin release, so HA may play a role in the etiology of depression. Results We compare model predictions to classical physiological experiments on HA half-life, the concentration of brain HA after histidine loading, and brain HA after histidine is dramatically increased or decreased in the diet. The model predictions are also consistent with in vivo experiments in which extracellular HA is measured, using Fast Scan Cyclic Voltammetry, in the premammillary nucleus (PM) after a 2 s antidromic stimulation of the TM, both without and in the presence of the H 3 autoreceptor antagonist thioperamide. We show that the model predicts well the temporal behavior of HA in the extracellular space over 30 s in both experiments. Conclusions Our ability to measure in vivo histamine dynamics in the extracellular space after stimulation presents a real opportunity to understand brain function and control. The observed extracellular dynamics depends on synthesis, storage, neuronal firing, release, reuptake, glial cells, and control by autoreceptors, as well as the behavioral state of the animal (for example, depression) or the presence of neuroinflammation. In this complicated situation, the mathematical model will be useful for interpreting data and conducting in silico experiments to understand causal mechanisms. And, better understanding can suggest new therapeutic drug targets.

The mechanisms that control extracellular serotonin levels in vivo are not well-defined. This shortcoming makes it very challenging to diagnose and treat the many psychiatric disorders in which serotonin is implicated. Fast-scan cyclic voltammetry (FSCV) can measure rapid serotonin release and reuptake events but cannot report critically important ambient serotonin levels. In this Article, we use fast-scan controlled adsorption voltammetry (FSCAV), to measure serotonin’s steady-state, extracellular chemistry. We characterize the “Jackson” voltammetric waveform for FSCAV and show highly stable, selective, and sensitive ambient serotonin measurements in vitro. In vivo, we report basal serotonin levels in the CA2 region of the hippocampus as 64.9 ± 2.3 nM (n = 15 mice, weighted average ± standard error). We electrochemically and pharmacologically verify the selectivity of the serotonin signal. Finally, we develop a statistical model that incorporates the uncertainty in in vivo measurements, in addition to electrode variability, to more critically analyze the time course of pharmacological data. Our novel method is a uniquely powerful analysis tool that can provide deeper insights into the mechanisms that control serotonin’s extracellular levels.

The precise regulation of synaptic dopamine (DA) content by the dopamine transporter (DAT) ensures the phasic nature of the DA signal, which underlies the ability of DA to encode reward prediction error, thereby driving motivation, attention, and behavioral learning. Disruptions to the DA system are implicated in a number of neuropsychiatric disorders, including attention deficit hyperactivity disorder (ADHD) and, more recently, Autism Spectrum Disorder (ASD). An ASD-associated de novo mutation in the SLC6A3 gene resulting in a threonine to methionine substitution at site 356 (DAT T356M) was recently identified and has been shown to drive persistent reverse transport of DA (i.e. anomalous DA efflux) in transfected cells and to drive hyperlocomotion in Drosophila melanogaster. A corresponding mutation in the leucine transporter, a DAT-homologous transporter, promotes an outward-facing transporter conformation upon substrate binding, a conformation possibly underlying anomalous dopamine efflux. Here we investigated in vivo the impact of this ASD-associated mutation on DA signaling and ASD-associated behaviors. We found that mice homozygous for this mutation display impaired striatal DA neurotransmission and altered DA-dependent behaviors that correspond with some of the behavioral phenotypes observed in ASD.

It is extremely challenging to chemically diagnose disorders of the brain. There is hence great interest in designing and optimizing tools for direct detection of chemical biomarkers implicated in neurological disorders to improve diagnosis and treatment. Tools that are capable of monitoring brain chemicals, neurotransmitters in particular, need to be biocompatible, perform with high spatiotemporal resolution, and ensure high selectivity and sensitivity. Recent advances in electrochemical methods are addressing these criteria; the resulting devices demonstrate great promise for in vivo neurotransmitter detection. None of these devices are currently used for diagnostic purposes, however these cutting-edge technologies are promising more sensitive, selective, faster, and less invasive measurements. Via this review we highlight significant technical advances and in vivo studies, performed in the last 5 years, that we believe will facilitate the development of diagnostic tools for brain disorders.

The subcallosal cingulate (SCC) region, or its rodent homologue the medial prefrontal cortex (mPFC), and midbrain dorsal raphe (DR) are crucial nodes of the widespread network implicated in emotional regulation. Stimulation of the SCC is being explored as a potential treatment for depression. Because modulation of the 5-HT system is the most common pharmacological means of treating depression, we sought to establish 5-HT's role in the mPFC-DR projection. Using anaesthetized mice, we recorded neuronal activity in 49 neurons of the DR before, during, and after high frequency stimulation (HFS) of the mPFC. The majority of DR cells (74%) significantly decreased firing rate during HFS (p

Rapid, selective detection of metals in complex samples remains an elusive goal that could provide critical analytical information for biological and environmental sciences and industrial waste management. Fast-scan cyclic voltammetry (FSCV) using carbon-fiber microelectrodes (CFMs) is an emerging technique for metal analysis with broad potential applicability because of its rapid response to changes in analyte concentration and minimal disturbance to the analysis medium. In this communication, we report the first effective application of covalently modified CFMs to achieve highly selective, subsecond Cu(II) measurements using FSCV. A two-part strategy is employed for maximum selectivity: Cu(II) binding is augmented by a covalently grafted ionophore, while binding of other metals is prevented by chemical blocking of nonselective surface adsorption sites. The resulting electrodes selectively detect Cu(II) in a complex medium comprising several interfering metals. Overall, this strategy represents a transformative innovation in real-time electrochemical detection of metal analytes.

Our understanding of the world around us has been greatly enhanced by our ability to detect and quantify analytes in the environment and in biology. It is highly beneficial to perform analysis at the site of interest as many situations do not afford the luxury of time required for samples to be collected and sent off for analysis. Some processes necessitate immediate detection to facilitate rapid action. As an example, harmful environmental phenomena can result from dynamic events such as the recent spate of tropical cyclones in the Caribbean and Southeastern region of the United States. These storms have mobilized nutrients, facilitating large-scale algal blooms that are damaging to marine ecosystems and human health. The rapid delivery of analytical information during such events is the key to quickly mitigating detrimental effects. Another example is analytically monitoring the quality of drinking water in the context of avoiding residential crises such as in Flint, MI where residents were exposed to h...