James C. Leiter

Neuroprotective mechanisms of cerium oxide nanoparticles in a mouse hippocampal brain slice model of ischemia.

Cerium oxide nanoparticles (nanoceria) are widely used as catalysts in industrial applications because of their potent free radical-scavenging properties. Given that free radicals play a prominent role in the pathology of many neurological diseases, we explored the use of nanoceria as a potential therapeutic agent for stroke. Using a mouse hippocampal brain slice model of cerebral ischemia, we show here that ceria nanoparticles reduce ischemic cell death by approximately 50%. The neuroprotective effects of nanoceria were due to a modest reduction in reactive oxygen species, in general, and ~15% reductions in the concentrations of superoxide (O(2)(•-)) and nitric oxide, specifically. Moreover, treatment with nanoceria markedly decreased (~70% reduction) the levels of ischemia-induced 3-nitrotyrosine, a modification to tyrosine residues in proteins induced by the peroxynitrite radical. These findings suggest that scavenging of peroxynitrite may be an important mechanism by which cerium oxide nanoparticles mitigate ischemic brain injury. Peroxynitrite plays a pivotal role in the dissemination of oxidative injury in biological tissues. Therefore, nanoceria may be useful as a therapeutic intervention to reduce oxidative and nitrosative damage after a stroke.

Amperometric detection of dopamine in vivo with an enzyme based carbon fiber microbiosensor.

We developed a novel implantable enzyme-based carbon fiber biosensor for in vivo monitoring of dopamine. The biosensor is fabricated using tyrosinase immobilized in a biocompatible matrix consisting of a biopolymer, chitosan and ceria-based metal oxides, deposited onto the surface of a carbon fiber microelectrode with a diameter of approximately 100 microm. Tyrosinase catalyzes the conversion of dopamine to o-dopaquinone, and the reduction of o-dopaquinone, which requires a low potential difference, was detected electrochemically. The role of each component in the sensing layer was systematically investigated in relation to the analytical performance of the biosensor. In its optimal configuration, the biosensor demonstrated a detection limit of 1 nM dopamine, a linear range of 5 orders of magnitude between 10 nM and 220 microM, a sensitivity of 14.2 nA x microM(-1), and good selectivity against ascorbic acid, uric acid, serotonin, norepinephrine, epinephrine, and 3,4-dihydroxy-l-phenylalanine (L-DOPA). The system provided continuous, real time monitoring of electrically stimulated dopamine release in the brain of an anesthetized rat. Levels of dopamine up to 1.69 microM were measured. This new implantable dopamine biosensor provides an alternative to fast scan cyclic voltammetry for in vivo monitoring of dopamine.

Dopamine efflux in the rat striatum evoked by electrical stimulation of the subthalamic nucleus: potential mechanism of action in Parkinson's disease

The precise mechanism whereby continuous high‐frequency electrical stimulation of the subthalamic nucleus ameliorates motor symptoms of Parkinsons disease is unknown. We examined the effects of high‐frequency stimulation of regions dorsal to and within the subthalamic nucleus on dopamine efflux in the striatum of urethane‐anaesthetized rats using constant potential amperometry. Complementary extracellular electrophysiological studies determined the activity of subthalamic nucleus neurons in response to similar electrical stimulation of the subthalamic nucleus. High‐frequency stimulation of the subthalamic nucleus increased action potential firing in the subthalamic nucleus only during the initial stimulation period and was followed by a cessation of firing over the remainder of stimulation. Electrical stimulation of the subthalamic nucleus with 15 pulses elicited stimulus‐time‐locked increases in striatal dopamine efflux with maximal peak effects occurring at 50 Hz frequency and 300 µA intensity. Extended subthalamic nucleus stimulation (1000 pulses at 50 Hz; 300 µA) elicited a similar peak increase in striatal dopamine efflux that was followed by a relatively lower steady‐state elevation in extracellular dopamine over the course of stimulation. In contrast, extended stimulation immediately adjacent and dorsal to the subthalamic nucleus resulted in an 11‐fold greater increase in dopamine efflux that remained elevated over the course of the stimulation. Immunohistochemical staining for tyrosine hydroxylase revealed catecholaminergic fibers running immediately dorsal to and through the subthalamic nucleus. Taken together, these results suggest that enhanced dopamine release within the basal ganglia may be an important mechanism whereby high‐frequency stimulation of the subthalamic nucleus improves motor symptoms of Parkinsons disease.

Low levels of dietary methylmercury inhibit growth and gonadal development in juvenile walleye (Stizostedion vitreum)

Mercury levels in the aquatic environment of North America have been increasing, raising the possibility that this highly toxic heavy metal might alter fish populations. Previous investigations have demonstrated toxic effects of mercury on teleost reproduction, but these findings were observed following unrealistically high exposures. In this study, we used concentrations frequently observed in North American lakes to investigate the effects of dietary methylmercury on growth, gonadal development, and plasma cortisol levels in juvenile walleye (Stizostedion vitreum). For a period of 6 months, two groups of walleye were reared on untainted catfish fillets, while two test groups were fed fillets injected with methylmercury, one group receiving 0.1 kg Hg g-’ food (low-mercury diet) and the other receiving 1 .O 1.18 Hg g 1 food (high-mercury diet). After the exposure period, fish fed the low- and high-mercury diets had mean body burdens of 0.254 f. 0.015 pg Hg g-l and 2.37 f 0.09 Fg Hg g-‘, respectively. Dietary mercury significantly impaired both growth and gonadal development in males, which was apparent as reduced fish length, weight, and gonadosomatic index. Testicular atrophy was observed in fish fed the mercury-tainted fillets, but was nonexistent in control animals. Mercury also suppressed plasma cortisol in juveniles (sexes combined). The findings of this study suggest that dietary methylmercury, at levels currently found in the aquatic environment, might reduce juvenile survival by impairing growth and immune function. Furthermore, these results suggest that methylmercury might also affect reproductive potential of teleosts by impairing testicular development in young.

Maternal nicotine depresses eupneic ventilation of neonatal rats

Maternal smoking is a risk factor for the sudden infant death syndrome (SIDS). We hypothesized that pre-natal exposure to nicotine would result in abnormalities of ventilatory activity in newborns. Neonatal rats which had been exposed to nicotine had significantly lower minute ventilation breathing air and hypoxic gas mixtures than did control animals. In both groups, anoxia elicited gasping which was equally effective in restoring eupnea. Maternal exposure to nicotine may result in a reduced metabolic rate and/or chronic hypoventilation in the newborn.

Inhibition of Monocarboxylate Transporter 2 in the Retrotrapezoid Nucleus in Rats: A Test of the Astrocyte–Neuron Lactate-Shuttle Hypothesis

The astrocyte-neuronal lactate-shuttle hypothesis posits that lactate released from astrocytes into the extracellular space is metabolized by neurons. The lactate released should alter extracellular pH (pHe), and changes in pH in central chemosensory regions of the brainstem stimulate ventilation. Therefore, we assessed the impact of disrupting the lactate shuttle by administering 100 μm α-cyano-4-hydroxy-cinnamate (4-CIN), a dose that blocks the neuronal monocarboxylate transporter (MCT) 2 but not the astrocytic MCTs (MCT1 and MCT4). Administration of 4-CIN focally in the retrotrapezoid nucleus (RTN), a medullary central chemosensory nucleus, increased ventilation and decreased pHe in intact animals. In medullary brain slices, 4-CIN reduced astrocytic intracellular pH (pHi) slightly but alkalinized neuronal pHi. Nonetheless, pHi fell significantly in both cell types when they were treated with exogenous lactate, although 100 μm 4-CIN significantly reduced the magnitude of the acidosis in neurons but not astrocytes. Finally, 4-CIN treatment increased the uptake of a fluorescent 2-deoxy-d-glucose analog in neurons but did not alter the uptake rate of this 2-deoxy-d-glucose analog in astrocytes. These data confirm the existence of an astrocyte to neuron lactate shuttle in intact animals in the RTN, and lactate derived from astrocytes forms part of the central chemosensory stimulus for ventilation in this nucleus. When the lactate shuttle was disrupted by treatment with 4-CIN, neurons increased the uptake of glucose. Therefore, neurons seem to metabolize a combination of glucose and lactate (and other substances such as pyruvate) depending, in part, on the availability of each of these particular substrates.

Deep brain stimulation of the nucleus accumbens reduces alcohol intake in alcohol-preferring rats

OBJECT The authors tested the hypothesis that deep brain stimulation (DBS) in the nucleus accumbens (NAcc) decreases alcohol intake in alcohol-preferring (P) rats after each animal has established a stable, large alcohol intake and after P rats with an established intake have been deprived of alcohol for 4-6 weeks. METHODS Bipolar stimulating electrodes were bilaterally placed in the NAcc using stereotactic coordinates. In the first study, P rats (9 animals) were allowed to establish a stable pattern of alcohol intake (about 5-7 g/day) over approximately 2 weeks, and the acute effects of DBS in the NAcc (140-150 Hz, 60-microsec pulse width, and 200-microA current intensity) on alcohol intake and alcohol preference were studied. Each animal acted as its own control and received 1 hour of DBS followed by 1 hour of sham-DBS or vice versa on each of 2 sequential days. The order of testing (sham-DBS vs DBS) was randomized. In the second study, each animal was allowed to establish a stable alcohol intake and then the animal was deprived of alcohol for 4-6 weeks. Animals received DBS (6 rats) or sham-DBS (5 rats) in the NAcc for 24 hours starting when alcohol was reintroduced to each animal. RESULTS Deep brain stimulation in the NAcc, as compared with a period of sham-DBS treatment in the same animals, acutely decreased alcohol preference. Furthermore, alcohol consumption and preference were significantly reduced in the DBS group compared with the sham treatment group during the first 24 hours that alcohol was made available after a period of forced abstinence. CONCLUSIONS The NAcc plays a key role in the rewarding and subsequent addictive properties of drugs of abuse in general and of alcohol in particular. Deep brain stimulation in the NAcc reduced alcohol consumption in P rats both acutely and after a period of alcohol deprivation. Therefore, DBS in the NAcc coupled with other neurophysiological measurements may be a useful tool in determining the role of the NAcc in the mesocorticolimbic reward circuit. Deep brain stimulation in the NAcc may also be an effective treatment for reducing alcohol consumption in patients who abuse alcohol and have not responded to other forms of therapy.

Neonatal maturation of the hypercapnic ventilatory response and central neural CO2 chemosensitivity

The ventilatory response to CO2 changes as a function of neonatal development. In rats, a ventilatory response to CO2 is present in the first 5 days of life, but this ventilatory response to CO2 wanes and reaches its lowest point around postnatal day 8. Subsequently, the ventilatory response to CO2 rises towards adult levels. Similar patterns in the ventilatory response to CO2 are seen in some other species, although some animals do not exhibit all of these phases. Different developmental patterns of the ventilatory response to CO2 may be related to the state of development of the animal at birth. The triphasic pattern of responsiveness (early decline, a nadir, and subsequent achievement of adult levels of responsiveness) may arise from the development of several processes, including central neural mechanisms, gas exchange, the neuromuscular junction, respiratory muscles and respiratory mechanics. We only discuss central neural mechanisms here, including altered CO2 sensitivity of neurons among the various sites of central CO2 chemosensitivity, changes in astrocytic function during development, the maturation of electrical and chemical synaptic mechanisms (both inhibitory and excitatory mechanisms) or changes in the integration of chemosensory information originating from peripheral and multiple central CO2 chemosensory sites. Among these central processes, the maturation of synaptic mechanisms seems most important and the relative maturation of synaptic processes may also determine how plastic the response to CO2 is at any particular age.

Mechanisms of pathogenesis in the Sudden Infant Death Syndrome.

The likely processes of the Sudden Infant Death Syndrome (SIDS) were identified many years ago (apnea, failed arousal, failed autoresuscitation, etc.). The neurophysiological basis of these processes and the neurophysiological reasons some infants die of SIDS and others do not are, however, only emerging now. We reviewed recent studies that have shed light on the way in which epidemiological risk factors, genetics, neurotransmitter receptor defects and neonatal cardiorespiratory reflex responses interact to lead to sudden death during sleep in a small number of normal appearing infants. As a result of this review and analysis, we hypothesize that the neurophysiological basis of SIDS resides in a persistence of fetal reflex responses into the neonatal period, amplification of inhibitory cardiorespiratory reflex responses and reduced excitatory cardiorespiratory reflex responses. The hypothesis we developed explores the ways in which multiple subtle abnormalities interact to lead to sudden death and emphasizes the difficulty of ante-mortem identification of infants at risk for SIDS, although identification of infants at risk remains an essential goal of SIDS research.

Deep Brain Stimulation Results in Local Glutamate and Adenosine Release: Investigation into the Role of Astrocytes

BACKGROUNDSeveral neurological disorders are treated with deep brain stimulation; however, the mechanism underlying its ability to abolish oscillatory phenomena associated with diseases as diverse as Parkinsons disease and epilepsy remain largely unknown. OBJECTIVETo investigate the role of specific neurotransmitters in deep brain stimulation and determine the role of non-neuronal cells in its mechanism of action. METHODSWe used the ferret thalamic slice preparation in vitro, which exhibits spontaneous spindle oscillations, to determine the effect of high-frequency stimulation on neurotransmitter release. We then performed experiments using an in vitro astrocyte culture to investigate the role of glial transmitter release in high-frequency stimulation-mediated abolishment of spindle oscillations. RESULTSIn this series of experiments, we demonstrated that glutamate and adenosine release in ferret slices was able to abolish spontaneous spindle oscillations. The glutamate release was still evoked in the presence of the Na+ channel blocker tetrodotoxin, but was eliminated with the vesicular H+-ATPase inhibitor bafilomycin and the calcium chelator 2-bis(2-aminophenoxy)-ethane-N,N,N′,N′-tetraacetic acid tetrakis acetoxymethyl ester. Furthermore, electrical stimulation of purified primary astrocytic cultures was able to evoke intracellular calcium transients and glutamate release, and bath application of 2-bis (2-aminophenoxy)-ethane-N,N,N′,N′-tetraacetic acid tetrakis acetoxymethyl ester inhibited glutamate release in this setting. CONCLUSIONVesicular astrocytic neurotransmitter release may be an important mechanism by which deep brain stimulation is able to achieve clinical benefits.