Scalable Nanostructured Carbon Electrode Arrays for Enhanced Dopamine Detection.

Dopamine is a neurotransmitter that modulates arousal and motivation in humans and animals. It plays a central role in the brain "reward" system. Its dysregulation is involved in several debilitating disorders such as addiction, depression, Parkinson's disease, and schizophrenia. Dopamine neurotransmission and its reuptake in extracellular space takes place with millisecond temporal and nanometer spatial resolution. Novel nanoscale electrodes are needed with superior sensitivity and improved spatial resolution to gain an improved understanding of dopamine dysregulation. We report on a scalable fabrication of dopamine neurochemical probes of a nanostructured glassy carbon that is smaller than any existing dopamine sensor and arrays of more than 6000 nanorod probes. We also report on the electrochemical dopamine sensing of the glassy carbon nanorod electrode. Compared with a carbon fiber, the nanostructured glassy carbon nanorods provide about 2× higher sensitivity per unit area for dopamine sensing and more than 5× higher signal per unit area at low concentration of dopamine, with comparable LOD and time response. These glassy carbon nanorods were fabricated by pyrolysis of a lithographically defined polymeric nanostructure with an industry standard semiconductor fabrication infrastructure. The scalable fabrication strategy offers the potential to integrate these nanoscale carbon rods with an integrated circuit control system and with other complementary metal oxide semiconductor (CMOS) compatible sensors.

Quantitation of levodopa and carbidopa in rat plasma by LC-MS/MS: The key role of ion-pairing reversed-phase chromatography.

A simple and selective bioanalytical method was developed for simultaneous determination of levodopa and carbidopa in rat plasma by LC-MS/MS. Levodopa and carbidopa are small polar molecules, posing challenges in the development of selective and efficient chromatography conditions. Perfluoropentanoic acid (PFPA), a volatile ion-pairing agent, was utilized to enhance chromatographic characteristics of both compounds in the reversed-phase mechanism. The ion-pairing chromatography played an essential role in mitigating matrix effects and achieving adequate separation between interfering background peaks and those of the analytes of interest, especially for levodopa. A 96-well based, automated liquid-liquid extraction, via the use Hamilton NIMBUS liquid handlers, was developed. Butyl alcohol, when mixed with ethyl acetate, greatly increased the recovery of both levodopa and carbidopa. The addition of PFPA further enhanced recovery for both analytes. Sodium metabisulfite, an antioxidant, was used to stabilize levodopa and carbidopa in rat plasma. The method was validated in the ranges of 50-10,000ng/mL and 25-5000ng/mL for levodopa and carbidopa, respectively, using levodopa-d3 and carbidopa-d3 as internal standards. The validated method was successfully applied to analyze rat plasma samples from in-life studies.

Phthalocyanine Doped Metal Oxide Nanoparticles on Multiwalled Carbon Nanotubes Platform for the detection of Dopamine.

The electrocatalytic properties of metal oxides (MO = Fe3O4, ZnO) nanoparticles doped phthalocyanine (Pc) and functionalized MWCNTs, decorated on glassy carbon electrode (GCE) was investigated. Successful synthesis of the metal oxide nanoparticles and the MO/Pc/MWCNT composite were confirmed using UV-Vis, EDX, XRD and TEM techniques. Successful modification of GCE with the MO and their composite was also confirmed using cyclic voltammetry (CV) technique. GCE-MWCNT/ZnO/29H,31H-Pc was the best electrode towards DA detection with very low detection limit (0.75 µM) which compared favourably with literature, good sensitivity (1.45 µA/µM), resistance to electrode fouling, and excellent ability to detect DA without interference from AA signal. Electrocatalytic oxidation of DA on GCE-MWCNT/ZnO/29H,31H-Pc electrode was diffusion controlled but characterized with some adsorption of electro-oxidation reaction intermediates products. The fabricated sensors are easy to prepare, cost effective and can be applied for real sample analysis of dopamine in drug composition. The good electrocatalytic properties of 29H,31H-Pc and 2,3-Nc were related to their (quantum chemically derived) frontier molecular orbital energies and global electronegativities. The better performance of 29H,31H-Pc than 2,3-Nc in aiding electrochemical oxidation of DA might be due to its better electron accepting ability, which is inferred from its lower ELUMO and higher χ.

Postmortem memantine concentration in a non-intoxication case, and the possibility of postmortem redistribution: A case report.

In this case study, we measured the concentration of memantine in the heart blood, peripheral blood, urine, liver, thigh muscle, and subcutaneous fat of a 64-year-old woman who was prescribed memantine for early-onset Alzheimer's disease. She died in hospital after an altercation with her husband. Cause of death was clearly not drug intoxication or overdose, so we investigated the postmortem redistribution (PMR) of memantine in the various tissues and blood ratios of the postmortem samples. Memantine concentrations detected were 1.31 µg/mL in the peripheral blood, 3.95 µg/mL in central blood, 2.09 µg/mL in the urine, 25.54 µg/g in the liver, 1.16 µg/g in the thigh muscle and 2.13 µg/g in the subcutaneous fat. In all samples, the concentrations were higher than the accepted therapeutic range (which is approximately 0.09-0.15 µg/mL). The central blood to peripheral blood (C/P) memantine ratio was 3.01 while the liver to peripheral blood (L/P) ratio was 19.5. It is documented that a C/P ratio exceeding 2 and L/P ratio exceeding 20 highlight a propensity for significant PMR. Although this is a single case study, our data suggest that memantine exhibits PMR. Additionally, a lowered pH was found in peripheral blood (pH 6.2) and central blood (pH 6.1). This postmortem reduction in blood pH may also promote the PMR of memantine. Because there is very little available postmortem toxicological data on memantine, our case study will serve as a foundation to assist in future forensic investigations.

Palladium nanoparticles decorated on activated fullerene modified screen printed carbon electrode for enhanced electrochemical sensing of dopamine.

In the present work, an enhanced electrochemical sensor for dopamine (DA) was developed based on palladium nanoparticles decorated activated fullerene-C60 (AC60/PdNPs) composite modified screen printed carbon electrode (SPCE). The scanning electron microscopy and elemental analysis confirmed the formation of PdNPs on AC60. The fabricated AC60/PdNPs composite modified electrode exhibited an enhanced electrochemical response to DA with a lower oxidation potential than that of SPCE modified with PdNPs and C60, indicating the excellent electrooxidation behavior of the AC60/PdNPs composite modified electrode. The electrochemical studies confirmed that the electrooxidation of DA at the composite electrode is a diffusion controlled electrochemical process. The differential pulse voltammetry was employed for the determination of DA; under optimum conditions, the electrochemical oxidation signal of DA increased linearly at the AC60/PdNPs composite from 0.35 to 133.35 µM. The limit of detection was found as 0.056 µM with a sensitivity of 4.23 µA µM(-1) cm(-2). The good recovery of DA in the DA injection samples further revealed the good practicality of AC60/PdNPs modified electrode.

Highly sensitive reduced graphene oxide microelectrode array sensor.

Reduced graphene oxide (rGO) has been fabricated into a microelectrode array (MEA) using a modified nanoimprint lithography (NIL) technique. Through a modified NIL process, the rGO MEA was fabricated by a self-alignment of conducting Indium Tin Oxide (ITO) and rGO layer without etching of the rGO layer. The rGO MEA consists of an array of 10µm circular disks and microelectrode signature has been found at a pitch spacing of 60µm. The rGO MEA shows a sensitivity of 1.91nAµm(-1) to dopamine (DA) without the use of mediators or functionalization of the reduced graphene oxide (rGO) active layer. The performance of rGO MEA remains stable when tested under highly resistive media using a continuous flow set up, as well as when subjecting it to mechanical stress. The successful demonstration of NIL for fabricating rGO microelectrodes on flexible substrate presents a route for the large scale fabrication of highly sensitive, flexible and thin biosensing platform.

Enhanced electrochemiluminescence sensor for detecting dopamine based on gold nanoflower@graphitic carbon nitride polymer nanosheet-polyaniline hybrids.

In this work, an enhanced electrochemiluminescence (ECL) sensor based on gold nanoflower@graphitic carbon nitride polymer nanosheet-polyaniline hybrids (AuNF@g-C3N4-PANI) was prepared for the detection of dapamine (DA). First, the bulk g-C3N4 was prepared through polymerizing melamine under 600 °C. And then the g-C3N4 nanosheet was obtained by ultrasonication-assisted liquid exfoliation of bulk g-C3N4. Finally, polyaniline (PANI) and gold nanoflowers (AuNFs) were successively formed on the g-C3N4 nanosheet through an in situ synthesis method. The resulting AuNF@g-C3N4-PANI hybrids were modified onto the surface of glassy carbon electrode to achieve a sensor (AuNF@g-C3N4-PANI/GCE) for detecting dopamine. Under the optimal conditions, the ECL signal increased linearly with the concentration of dopamine. The linear range of 5.0 × 10(-9) to 1.6 × 10(-6) M was obtained, while the detection limit was 1.7 × 10(-9) M. The prepared sensor exhibited a low detection limit and high sensitivity for the determination of dopamine. The combination of g-C3N4 nanosheet, PANI and AuNF would provide a new opportunity for the ECL sensor.

Synthesis of ZnO nanorods and their application in the construction of a nanostructure-based electrochemical sensor for determination of levodopa in the presence of carbidopa.

A novel carbon paste electrode modified with ZnO nanorods and 5-(4'-amino-3'-hydroxy-biphenyl-4-yl)-acrylic acid (3,4'-AAZCPE) was fabricated. The electrochemical study of the modified electrode, as well as its efficiency for the electrocatalytic oxidation of levodopa, is described. The electrode was employed to study the electrocatalytic oxidation of levodopa, using cyclic voltammetry (CV), chronoamperometry (CHA), and square-wave voltammetry (SWV) as diagnostic techniques. It has been found that the oxidation of levodopa at the surface of the modified electrode occurs at a potential of about 370 mV less positive than that of an unmodified carbon paste electrode. The SWV results exhibit a linear dynamic range from 1.0 × 10(-7) M to 7.0 × 10(-5) M and a detection limit of 3.5 × 10(-8) M for levodopa. In addition, this modified electrode was used for the simultaneous determination of levodopa and carbidopa. Finally, the modified electrode was used for the determination of levodopa and carbidopa in some real samples.

Potentiometric sensors doped with biomolecules as a new approach to small molecule/biomolecule binding kinetics analysis.

The most successful binding kinetics analysis systems at this moment include surface plasmon resonance (SPR), quartz microcrystal balance (QMB) and surface acoustic wave (SAW). Although these are powerful methods, they generally are complex, expensive and require the use of monolayers. Here, we report on potentiometric sensors as an inexpensive and simple alternative to do binding kinetics analysis between small molecules in solution and biomolecules (covalently) attached in a biopolymer sensor coating layer. As an example, dopamine and an anti-dopamine aptamer were used as the small molecule and the biomolecule respectively. Binding between both follows a Langmuir adsorption type model and creates a surface potential. The system operates in Flow Injection Analysis mode (FIA). Besides being an interesting new binding kinetics tool, the approach allows systematic design of potentiometric biosensors (in the present study a dopamine sensor), and gives new insights into the functioning of ion-selective electrodes (ISE's).

Carbon nanotube multi-electrode array chips for noninvasive real-time measurement of dopamine, action potentials, and postsynaptic potentials.

Multi-electrode arrays (MEAs) can be used for noninvasive, real-time, and long-term recording of electrophysiological activity and changes in the extracellular chemical microenvironment. Neural network organization, neuronal excitability, synaptic and phenotypic plasticity, and drug responses may be monitored by MEAs, but it is still difficult to measure presynaptic activity, such as neurotransmitter release, from the presynaptic bouton. In this study, we describe the development of planar carbon nanotube (CNT)-MEA chips that can measure both the release of the neurotransmitter dopamine as well as electrophysiological responses such as field postsynaptic potentials (fPSPs) and action potentials (APs). These CNT-MEA chips were fabricated by electroplating the indium-tin oxide (ITO) microelectrode surfaces. The CNT-plated ITO electrode exhibited electrochemical response, having much higher current density compared with the bare ITO electrode. Chronoamperometric measurements using these CNT-MEA chips detected dopamine at nanomolar concentrations. By placing mouse striatal brain slices on the CNT-MEA chip, we successfully measured synaptic dopamine release from spontaneous firings with a high S/N ratio of 62. Furthermore, APs and fPSPs were measured from cultured hippocampal neurons and slices with high temporal resolution and a 100-fold greater S/N ratio. Our CNT-MEA chips made it possible to measure neurotransmitter dopamine (presynaptic activities), postsynaptic potentials, and action potentials, which have a central role in information processing in the neuronal network. CNT-MEA chips could prove useful for in vitro studies of stem cell differentiation, drug screening and toxicity, synaptic plasticity, and pathogenic processes involved in epilepsy, stroke, and neurodegenerative diseases.

Sensitive and selective electrochemical detection of dopamine using an electrode modified with carboxylated carbonaceous spheres.

A highly sensitive and selective electrochemical sensor of dopamine (DA) has been developed by employing carboxylated carbonaceous spheres to modify glassy carbon electrodes (GCEs). Scanning electron microscopy (SEM) and Fourier transform infrared (FT-IR) spectroscopy were used to characterize as-prepared carbonaceous spheres. The results show that the diameter of carboxylated carbonaceous spheres is uniformly 500 nm and that their surfaces mainly expose carboxyl groups with negative charges. Electrochemical measurements demonstrate that carboxylated carbonaceous spheres greatly improve the accumulation of positively charged dopamine, leading to good sensing performance on a modified GCE. Through applying the differential pulse voltammetric approach, linear calibration curves were obtained in a range of about 0.1 to 40 µM with a detection limit down to 30 nM. Furthermore, depending on the charge-based discrimination, the modified electrode displays good selective detection of DA and reliable anti-interference to UA and glucose besides a weak and negligible response to AA. Therefore, the carboxylated carbonaceous sphere introduced here is a good candidate to develop electrochemical sensors for the sensitive and selective detection of DA.

An 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid)-immobilized electrode for the simultaneous detection of dopamine and uric acid in the presence of ascorbic acid.

An 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS)-immobilized carbon nanotube (CNT) electrode was used to simultaneously detect dopamine (DA) and uric acid (UA) in the presence of ascorbic acid (AA) with differential pulse voltammetry. When ABTS was immobilized onto the CNT electrode in the presence of DA, UA and 100 µM AA, the sensitivity to DA increased from 0.600 (±0.013) to 1.334 (±0.010) µA/µM in the concentration ranges of 0.90-10 µM and 1.87-20 µM, respectively, and the sensitivity to UA increased from 0.030 (±0.005) to 0.078 (±0.006) µA/µM in the concentration ranges of 2.16-240 µM and 3.07-400 µM, respectively. These findings demonstrate that the ABTS-immobilized CNT electrode attained a higher sensitivity to UA and also a wider linear range of concentrations.

UHPLC/ESI-Q-TOF-MS method for the measurement of dopamine in rodent striatal tissue: a comparative effects of intranasal administration of ropinirole solution over nanoemulsion.

An ultra high performance liquid chromatography-electrospray ionization-tandem mass spectrometric method (UHPLC/ESI-Q-TOF-MS) for the analysis of dopamine (DA) in Wistar rat brain homogenate has been developed and validated. The chromatographic separation was achieved on a Waters ACQUITY UPLC™ BEH C18 (100.0 mm × 2.1 mm; 1.7 µm) column using isocratic mobile phase, consisting of acetonitrile and Formic acid (0.1% w/v) (10: 90; v/v), at a flow rate of 0.15 ml min(-1) . The transitions occurred at m/z 154.04 → 137.006 for DA, and m/z 184.204 → 166.08 for the internal standard. The recovery of the analytes from Wistar rat brain homogenate was optimized using liquid-liquid extraction technique (LLE) in ethyl acetate. The total run time was 3.5 min and the elution of DA occurred at 1.44 ± 0.05 min. The linear dynamic range was established over the concentration range 75-750 ng mL(-1) (r(2) ; 0.9921 ± 0.0005) for DA. The intra-assay and inter-assay accuracy in terms of % CV was in the range 0.73-2.80. The lower limit of detection (LOD) and quantitation (LOQ) for DA was 0.278 and 0.844 ng mL(-1) , respectively. Analytes were stable under various conditions (in autosampler, during freeze-thaw, at room temperature, and under deep-freeze conditions). The developed method was successfully applied for in vivo profiling in rodents.

Application of ionic liquid-TiO2 nanoparticle modified carbon paste electrode for the voltammetric determination of benserazide in biological samples.

An ionic liquid-TiO2 nanoparticle modified carbon paste electrode (IL-TiO2/CPE) was used as a fast and sensitive tool for the investigation of the electrochemical oxidation of benserazide using voltammetry. This modified electrode has been fabricated using hydrophilic ionic liquid (n-hexyl-3-methylimidazolium hexafluoro phosphate) as a binder. The modified electrode offers a considerable improvement in voltammetric sensitivity toward benserazide, compared to the bare electrode. Using differential pulse voltammetry (DPV), the electrocatalytic oxidation peak current of benserazide shows a linear calibration curve in the range of 1.0-600 µmol L(-1) benserazide. The limit of detection was equal to 0.4 µmol L(-1). The relative standard deviation (RSD%) for eight successive assays of 10 µmol L(-1) benserazide was 1.1%. Finally, the proposed method was successfully applied to the determination of benserazide in real samples such as blood serum and urine.

Poly(methylene blue) functionalized graphene modified carbon ionic liquid electrode for the electrochemical detection of dopamine.

An ionic liquid 1-butylpyridinium hexafluorophosphate based carbon ionic liquid electrode (CILE) was used as the substrate electrode and a poly(methylene blue) (PMB) functionalized graphene (GR) composite film was co-electrodeposited on CILE surface by cyclic voltammetry. The PMB-GR/CILE exhibited better electrochemical performances with higher conductivity and lower electron transfer resistance. Electrochemical behavior of dopamine (DA) was further investigated by cyclic voltammetry and a pair of well-defined redox peaks appeared with the peak-to-peak separation (ΔE(p)) as 0.058V in 0.1 mol L(-1) pH 6.0 phosphate buffer solution, which proved a fast quasi-reversible electron transfer process on the modified electrode. Electrochemical parameters of DA on PMB-GR/CILE were calculated with the electron transfer number as 1.83, the charge transfer coefficients as 0.70, the apparent heterogeneous electron transfer rate constant as 1.72 s(-1) and the diffusional coefficient (D) as 3.45×10(-4) cm(2) s(-1), respectively. Under the optimal conditions with differential pulse voltammetric measurement, the linear relationship between the oxidation peak current of DA and its concentration was obtained in the range from 0.02 to 800.0 µmol L(-1) with the detection limit as 5.6 nmol L(-1) (3σ). The coexisting substances exhibited no interference and PMB-GR/CILE was applied to the detection of DA injection samples and human urine samples with satisfactory results.

Blood-brain barrier unlocked.

The brain is protected by a physiological blood-brain barrier (BBB) against toxins and some metabolites circulating in the blood. At the same time, the BBB limits penetration into the brain of many neuroactive drugs. Efficient ways to increase BBB permeability for delivery of drugs of different chemical nature into the brain are unknown. This work deals with delivery into the brain of 10(-2) M dopamine, a substance that does not penetrate the BBB under normal circumstances. It was studied in two independent experiments: (i) penetration of (3)H-labeled dopamine from its mixture with 10(-5) M H2O2 into hypothalamus and striatum structures of intact rat brain, and (ii) effect of unlabeled dopamine from a mixture with H(2)O(2) on the rat motor activity in a haloperidol catalepsy model. It was shown that (i) at the third minute after nasal application of the dopamine + H(2)O(2) mixture, the dopamine level increases 45-fold in the hypothalamus and almost 30-fold in the striatum and (ii) motility of animals in the catalepsy haloperidol model is recovered 90 sec after intranasal introduction of dopamine. No such effects were observed after replacement of H(2)O(2) by 0.9% NaCl solution. Thus, it was shown on the example of dopamine that its introduction into the nasal cavity simultaneously with H(2)O(2) provides for rapid delivery of the drug into the brain. These results expand our knowledge concerning the biological role of exoROS in modulating BBB permeability and may contribute to the development of a new therapeutic strategy for neurological diseases.

A novel electrochemical sensor for determination of dopamine based on AuNPs@SiO2 core-shell imprinted composite.

A novel core-shell composite of gold nanoparticles (AuNPs) and SiO(2) molecularly imprinted polymers (AuNPs@SiO(2)-MIPs) was synthesized through sol-gel technique and applied as a molecular recognition element to construct an electrochemical sensor for determination of dopamine (DA). Compared with previous imprinting recognition, the main advantages of this strategy lie in the introduction and combination of AuNPs and biocompatible porous sol-gel material (SiO(2)). The template molecules (DA) were firstly adsorbed at the AuNPs surface due to their excellent affinity, and subsequently they were further assembled onto the polymer membrane through hydrogen bonds and π-π interactions formed between template molecules and silane monomers. Cyclic voltammetry (CV) was carried out to extract DA molecules from the imprinted membrane, and as a result, DA could be rapidly and effectively removed. The AuNPs@SiO(2)-MIPs was characterized by ultraviolet visible (UV-vis) absorbance spectroscopy, transmission electron microscope (TEM) and Fourier transform infrared spectrometer (FT-IR). The prepared AuNPs@SiO(2)-MIPs sensor exhibited not only high selectivity toward DA in comparison to other interferents, but also a wide linear range over DA concentration from 4.8 × 10(-8) to 5.0 × 10(-5)M with a detection limit of 2.0 × 10(-8)M (S/N=3). Moreover, the new electrochemical sensor was successfully applied to the DA detection in dopamine hydrochloride injection and human urine sample, which proved that it was a versatile sensing tool for the selective detection of DA in real samples.

Efficient one-pot synthesis of molecularly imprinted silica nanospheres embedded carbon dots for fluorescent dopamine optosensing.

A new type of eco-friendly molecularly imprinted polymer (MIP) was synthesized through an efficient one-pot room-temperature sol-gel polymerization and applied as a molecular recognition element to construct dopamine (DA) fluorescence (FL) optosensor. Highly luminescent carbon dots (CDs) were firstly synthesized via a one-step reaction in organosilane, and their surface were anchored with MIP matrix (CDs@MIP). The resulting composite of a synergetic combination of CDs with MIP showed high photostability and template selectivity. Moreover, the composite allowed a highly sensitive determination of DA via FL intensity decreasing when removal of the original templates. The new MIP-based DA sensing protocol was applied to detect DA concentration in aqueous solution, the relative FL intensity of CDs@MIP decreased linearly with the increasing DA in the concentration range of 25-500nM with a detection limit (3σ) of 1.7 nM. Furthermore, the proposed method was successfully intended for the determination of trace DA in human urine samples without the interference of other molecules and ions.

Determination of dopamine in pharmaceutical formulation using enhanced luminescence from europium complex.

Biologically important compound dopamine plays an important role in the central and peripheral nervous systems. Insufficient dopamine level due to the loss of dopamine producing cells may lead to disease called Schizophrenia and Parkinson's disease. Hence, a simple and fast detection of dopamine is necessary to study in the fields of neurophysiology and clinical medicine. An enhanced fluorimetric determination of dopamine in the presence of ascorbic acid is achieved using photoluminescence of europium complex, Eu(III)-dipicolinic acid. In order to obtain better responses, several operational parameters have been investigated. Under the optimum conditions, the method showed good stability and reproducibility. The application of this method for the determination of dopamine neurotransmitters was satisfactory. Linear response was found down to 3.0 × 10(-7)M with limit of detection 1.0 × 10(-8)M. The relative standard deviation was found to be 3.33% from 20 independent measurements for 1.0 × 10(-5)M of dopamine.

Severe postnatal iron deficiency alters emotional behavior and dopamine levels in the prefrontal cortex of young male rats.

Iron deficiency in early human life is associated with abnormal neurological development. The objective of this study was to evaluate the effect of postnatal iron deficiency on emotional behavior and dopaminergic metabolism in the prefrontal cortex in a young male rodent model. Weanling, male, Sprague-Dawley rats were fed standard nonpurified diet (220 mg/kg iron) or an iron-deficient diet (2-6 mg/kg iron). After 1 mo, hematocrits were 0.42 ± 0.0043 and 0.16 ± 0.0068 (mean ± SEM; P < 0.05; n = 8), liver nonheme iron concentrations were 2.3 ± 0.24 and 0.21 ± 0.010 µmol/g liver (P < 0.05; n = 8), and serum iron concentrations were 47 ± 5.4 and 23 ± 7.1 µmol/L (P < 0.05; n = 8), respectively. An elevated plus maze was used to study emotional behavior. Iron-deficient rats displayed anxious behavior with fewer entries and less time spent in open arms compared to control rats (0.25 ± 0.25 vs. 1.8 ± 0.62 entries; 0.88 ± 0.88 vs. 13 ± 4.6 s; P < 0.05; n = 8). Iron-deficient rats also traveled with a lower velocity in the elevated plus maze (1.2 ± 0.15 vs. 1.7 ± 0.12 cm/s; P < 0.05; n = 8), behavior that reflected reduced motor function as measured on a standard accelerating rotarod device. Both the time on the rotarod bar before falling and the peak speed attained on rotarod by iron-deficient rats were lower than control rats (156 ± 12 vs. 194 ± 12 s; 23 ± 1.5 vs. 28 ± 1.6 rpm; P < 0.05; n = 7-8). Microdialysis experiments showed that these behavioral effects were associated with reduced concentrations of extracellular dopamine in the prefrontal cortex of the iron-deficient rats (79 ± 7.0 vs. 110 ± 14 ng/L; P < 0.05; n = 4). Altered dopaminergic signaling in the prefrontal cortex most likely contributes to the anxious behavior observed in young male rats with severe iron deficiency.