Biosensors for brain trauma and dual laser doppler flowmetry: enoxaparin simultaneously reduces stroke-induced dopamine and blood flow while enhancing serotonin and blood flow in motor neurons of brain, in vivo.

Neuromolecular Imaging (NMI) based on adsorptive electrochemistry, combined with Dual Laser Doppler Flowmetry (LDF) is presented herein to investigate the brain neurochemistry affected by enoxaparin (Lovenox(®)), an antiplatelet/antithrombotic medication for stroke victims. NMI with miniature biosensors enables neurotransmitter and neuropeptide (NT) imaging; each NT is imaged with a response time in milliseconds. A semiderivative electronic reduction circuit images several NT's selectively and separately within a response time of minutes. Spatial resolution of NMI biosensors is in the range of nanomicrons and electrochemically-induced current ranges are in pico- and nano-amperes. Simultaneously with NMI, the LDF technology presented herein operates on line by illuminating the living brain, in this example, in dorso-striatal neuroanatomic substrates via a laser sensor with low power laser light containing optical fiber light guides. NMI biotechnology with BRODERICK PROBE(®) biosensors has a distinct advantage over conventional electrochemical methodologies both in novelty of biosensor formulations and on-line imaging capabilities in the biosensor field. NMI with unique biocompatible biosensors precisely images NT in the body, blood and brain of animals and humans using characteristic experimentally derived half-wave potentials driven by oxidative electron transfer. Enoxaparin is a first line clinical treatment prescribed to halt the progression of acute ischemic stroke (AIS). In the present studies, BRODERICK PROBE(®) laurate biosensors and LDF laser sensors are placed in dorsal striatum (DStr) dopaminergic motor neurons in basal ganglia of brain in living animals; basal ganglia influence movement disorders such as those correlated with AIS. The purpose of these studies is to understand what is happening in brain neurochemistry and cerebral blood perfusion after causal AIS by middle cerebral artery occlusion in vivo as well as to understand consequent enoxaparin and reperfusion effects actually while enoxaparin is inhibiting blood clots to alleviate AIS symptomatology. This research is directly correlated with the medical and clinical needs of stroke victims. The data are clinically relevant, not only to movement dysfunction but also to the depressive mood that stroke patients often endure. These are the first studies to image brain neurotransmitters while any stroke medications, such as anti-platelet/anti-thrombotic and/or anti-glycoprotein are working in organ systems to alleviate the debilitating consequences of brain trauma and stroke/brain attacks.

Laurate Biosensors Image Brain Neurotransmitters In Vivo: Can an Antihypertensive Medication Alter Psychostimulant Behavior?

Neuromolecular Imaging (NMI) with novel biosensors enables the selective detection of neurotransmitters in vivo within seconds, on line and in real time. Biosensors remain in place for continuing studies over a period of months. This biotechnological advance is based on conventional electrochemistry; the biosensors detect neurotransmitters by electron transfer. Simply stated, biosensors adsorb electrons from each neurotransmitter at specific oxidation potentials; the current derived from electron transfer is proportional to neurotransmitter concentration. Selective electron transfer properties of these biosensors permit the imaging of neurotransmitters, metabolites and precursors. The novel BRODERICK PROBE® biosensors we have developed, differ in formulation and detection capabilities from biosensors/electrodes used in conventional electrochemistry/ voltammetry. In these studies, NMI, specifically, the BRODERICK PROBE® laurate biosensor images neurotransmitter signals within mesolimbic neuronal terminals, nucleus accumbens (NAc); dopamine (DA), serotonin (5-HT), homovanillic acid (HVA) and Ltryptophan (L-TP) are selectively imaged. Simultaneously, we use infrared photobeams to monitor open-field movement behaviors on line with NMI in the same animal subjects. The goals are to investigate integrated neurochemical and behavioral effects of cocaine and caffeine alone and co-administered and further, to use ketanserin to decipher receptor profiles for these psychostimulants, alone and co-administered. The rationale for selecting this medication is: ketanserin (a) is an antihypertensive and cocaine and caffeine produce hypertension and (b) acts at 5-HT2A/2C receptors, prevalent in NAc and implicated in hypertension and cocaine addiction. Key findings are: (a) the moderate dose of caffeine simultaneously potentiates cocaine's neurochemical and behavioral responses. (b) ketanserin simultaneously inhibits cocaine-increased DA and 5-HT release in NAc and open-field behaviors and (c) ketanserin inhibits 5-HT release in NAc and open-field behaviors produced by caffeine, but, surprisingly, acts to increase DA release in NAc. Importantly, the latter effect may be a possible adverse effect of the moderate dose of caffeine in hypertensive patients. Thus, an antihypertensive medication is shown here to play a role in inhibiting brain reward possibly via antihypertensive mechanisms at DA and 5-HT receptor subtypes within DA motor neurons. An explanatory note for the results obtained, is the role likely played by the G Protein Receptor Complex (GPRC) family of proteins. Empirical evidence shows that GPRC dimers, heteromers and heterotrimers may cause cross-talk between distinct signalling cascade pathways in the actions of cocaine and caffeine. Ligand-directed functional selectivity, particularly for ketanserin, in addition to GPRCs, may also cause differential responses. The results promise new therapeutic strategies for drug addiction, brain reward and cardiovascular medicine.

Monoamine and motor responses to cocaine are co-deficient in the Fawn-Hooded depressed animal model.

The Fawn-Hooded (FH) genetic animal model of depression continues to be of interest because the FH model has limited biochemical and immune function. The FH animal has an inherited trait, platelet storage pool deficiency (PSPD), an hemorrhagic disorder that is also a component of Chediak-Higashi syndrome (CHS). CHS is a pyrogenic infectious childhood disease; few patients live past the age of 20. Our hypothesis was that FH animals may exhibit different monoamine and motor responses to cocaine versus the Sprague-Dawley (SD) "normal" animal strain, which does not have the FH trait. Therefore, selective neuromolecular imaging (NMI) of the monoamines, dopamine (DA) and 5-HT within nucleus accumbens (NAcc) of behaving male FH versus SD rats was performed in vivo with BRODERICK PROBE sensors and a semiderivative voltammetric circuit. Each animal was placed in a faraday chamber and electrochemical signals were detected via a mercury commutator and flexible cable. Baseline values for neurotransmitters and behavior were derived during the last half-hour of habituation behavior. Release of DA and 5-HT was detected selectively, at separate oxidation potentials, within seconds, before and after intraperitoneal administration of the psychostimulant, cocaine (10 mg/kg). At the same time, frequencies of ambulations and central ambulations were separately monitored with infrared photobeams, which surrounded the faraday chamber. Data were compared by ANOVA analysis followed by Tukey's post hoc test. The data showed that (1) DA release in NAcc of behaving FH animals did not respond to cocaine; neither first hour nor second hour values significantly differed from baseline (both hours, p>0.05), whereas SD animals exhibited a significant increase in cocaine-induced DA release in NAcc (both hours, p<0.001). The ability for acute cocaine to increase DA release in NAcc was significantly greater in SD than in FH animals (p<0.001). (2) 5-HT release in NAcc of behaving FH animals was not significantly increased by cocaine (both hours, p>0.05), whereas 5-HT release in NAcc of SD animals was significantly increased after cocaine (both hours, p<0.001). The ability for acute cocaine to increase 5-HT release was significantly greater in SD than in FH animals (p<0.001). (3) Ambulations in the FH strain were modestly, yet significantly, enhanced after cocaine during both hours of study (p<0.05, p<0.001, respectively) as were ambulations in the SD strain. Nonetheless, the ability for acute cocaine to increase ambulations was significantly greater in SD than in FH animals in the first hour (p<0.001). (4) Central ambulations in the FH strain was not affected by cocaine (both hours, p>0.05), whereas SD animals showed a significant increase in central ambulatory activity in both hours of the cocaine study (p<0.001). The ability for acute cocaine to increase central ambulations was significantly greater in SD than in FH animals (p<0.001). Thus, this is the first study to determine in vivo the neurochemical response to acute cocaine in the behaving FH animal. Moreover, this is the first study to determine in vivo and simultaneously the neurochemical and behavioral response to acute cocaine in the FH strain in comparison with SD animals, a "normal" strain. Remarkable deficiencies in the ability for acute cocaine to alter neurochemistry and behavior in animals with the FH trait are shown. These studies emphasize the need to look differentially at cocaine effects in biochemically and immune-compromised subjects versus "normal" subjects.

Cytokines, stressors, and clinical depression: augmented adaptation responses underlie depression pathogenesis.

By influencing the central nervous system, cytokines, which regulate immune function innately and adaptively, may play a key role in mediating depression-like neuro-behavioral changes. However, the similarity between cytokine and stressor-effects in animal models raises a question about the degree to which behavioral and neurochemical outcomes of cytokine challenge represent depressive disorder per se. The present review attempts to illustrate the degree of overlap between cytokines and stressors with respect to their effects on neurochemistry and behavior in animal models. The review also shows how short-term effects of cytokine exposure in typical animals may be discerned from characteristics that might otherwise be described as depression-like. By comparing outcomes of immune challenge in typical rodent strains (e.g., Sprague-Dawley [SD], Wistar) and an accepted animal model of depression (e.g., Fawn Hooded [FH] rodent strain), differences between short-term effects of cytokines and depression-like characteristics in rodents are demonstrated. Additionally, because it is known that preexisting vulnerability to depression may affect outcomes of immune challenge, we further compare immunological, biochemical and behavioral effects of cytokines between SD and FH rodent strains. Interestingly, the acute neurochemical and behavioral effects of the cytokine interleukin 1alpha (IL-1alpha) reveal stressor-like responses during behavioral habituation in both strains, though this appears to a stronger degree in FH animals. Further, the subacute response to IL-1alpha vastly differed between strains, indicating differences in adaptive mechanisms. Thus, stressor-like effects of immune challenge, particularly in FH animals, provide validation for recent "cross-sensitization" models of depression pathogenesis that incorporate immune factors.

Interleukin 1alpha alters hippocampal serotonin and norepinephrine release during open-field behavior in Sprague-Dawley animals: differences from the Fawn-Hooded animal model of depression.

Detection of two biogenic amine neurotransmitters, serotonin (5-HT) and norepinephrine (NE) within the CA1 region of the hippocampus (HPC) of behaving male laboratory animals (Rattus norvegicus), was performed with miniature carbon sensors (BRODERICK PROBES) and in vivo semidifferential microvoltammetry after acute administration of the soluble immune factor, human recombinant, interleukin (IL) 1alpha (10 and 100 ng/kg i.p.). Two animal models were compared, i.e., (a) the Sprague-Dawley (SD) model, a strain neither biochemically nor immune-challenged and (b) the Fawn-Hooded (FH) model, a biochemically (5-HT-deficient) and immune-challenged animal. Open-field behaviors, locomotion (ambulations) and stereotypy (fine movements of sniffing and grooming) were monitored with infrared photobeams while 5-HT and NE were selectively and separately detected within seconds in real time. Subchronic studies were performed in the same animals 24 h later at which time no further drug was administered. Results from acute treatment studies showed that IL-1alpha altered HPC monoamines and behavior viz-a-viz habituation values (baseline) in the SD strain differently from those in the FH strain as follows: (1) although 5-HT release was significantly increased within CA1 region of HPC in both SD and FH strains (P<.0001), the extent of the HPC 5-HT increase in the 5-HT-deficient FH strain was significantly less than that of the SD strain at both doses (P<.0001). The subchronic studies showed that 5-HT release within the HPC in the SD strain significantly increased (135%) over drug treatment values (P<.001), whereas HPC 5-HT release in the FH strain remained the same as that seen in the acute drug treatment studies; the difference between strains for the subchronic study was also statistically significant (P<.01). (2) IL-1alpha significantly decreased HPC NE release in the SD strain (P<.0004) while IL-1alpha decreased HPC NE release in the FH strain only at the 10-ng/kg dose (P<.001); at the 100-ng/kg dose in the FH strain, NE rebounded towards baseline and increased 15% above baseline reaching statistical significance (P<.05). Subchronic studies in the SD strain showed a further decreased NE signal to 38% below baseline (P<.0001), whereas subchronic studies in the FH strain showed a significant increase in NE release (P<.02). The difference between strains in the subchronic NE studies was significant (P<.001). (3) Ambulations were increased after IL-1alpha administration in acute studies in both the SD and the FH strains, but the increase did not reach statistical significance, whereas in the subchronic studies, both strains exhibited significant increases as revealed by post hoc analyses (P<.05). There was a statistically significant difference between strains in acute studies (P<.001), whereas no significant differences between models were seen in ambulation behavior in subchronic studies. (4) Fine movements increased over baseline after IL-1alpha administration in both animal models in acute studies, however, results did not reach statistical significance, likely due to the episodic effect of IL-1alpha on movement behavior in both the SD and the FH strains. However, the SD strain showed a significant increase in fine movement behavior during the subchronic studies (P<.02). Significant differences in fine movements between animal models were not observed either acutely or in subchronic studies. In summary, the data show that immune modulation by IL-1alpha affects HPC neurochemistry and behavior in SD versus FH animal models differently and/or to different degrees. The data show that while the FH animal model is subsensitive to 5-HT agonists, 5-HT function can be stimulated. Comparison of genetically diverse animal models provides a reliable means to identify and discern cytokine-induced depressive versus stressor properties. Selective sensor technology provides a powerful tool as movement behavior is monitored and interpreted as a function of monoamine neurotransmission.

Acute and subacute effects of risperidone and cocaine on accumbens dopamine and serotonin release using in vivo microvoltammetry on line with open-field behavior.

In vivo microvoltammetry was used to detect dopamine (DA) and serotonin (5-HT) release from nucleus accumbens (NAcc) of freely moving, male, Sprague-Dawley laboratory rats, while animals' locomotor (forward ambulations) and stereotypic behavior (fine movements of sniffing and grooming) were monitored at the same time with infrared photobeams. Monoamine release mechanisms were determined by using a depolarization blocker (gamma-butyrolactone, gamma BL). Miniature carbon sensors (BRODERICK PROBES microelectrodes) smaller than a human hair were used in conjunction with a semidifferential electrochemical circuit to detect release of each monoamine in separate signals and within seconds. The purpose was to evaluate the neuropharmacology of the 5-HT(2)/DA(2) antagonist risperidone in its current therapeutic role as an atypical antipsychotic medication as well as in its potential role as pharmacotherapy for cocaine psychosis and withdrawal symptoms. Acute (single drug dose) and subacute (24-h follow-up studies in the same animal, no drug administration) studies were performed for each treatment group. The hypothesis for the present studies is derived from a growing body of evidence that cocaine-induced psychosis and schizophrenic psychosis share similar neurochemical and behavioral manifestations. Results showed that (1) Acute administration of risperidone (2 mg/kg sc) significantly increased DA and 5-HT release in NAcc above baseline (habituation) values (P<.001) while locomotion and stereotypy were virtually unaffected. In subacute studies, DA release did not differ from baseline (P>.05), whereas 5-HT release was significantly increased above baseline (P<.001). Locomotion increased over baseline but not to a significant degree, while stereotypy was significantly increased above baseline (P<.05). (2) Acute administration of cocaine (10 mg/kg ip) significantly increased both DA and 5-HT release above baseline (P<.001), while locomotion and stereotypy were significantly increased over baseline (P<.001). In subacute studies, DA decreased significantly below baseline (P<.001) and significant decreases in 5-HT release occurred at 15, 20, 50 and 55 min (P<.05). Behavior increased above baseline but did not reach a statistically significant degree. (3) Acute administration of risperidone/cocaine (2 mg/kg sc and 10 mg/kg ip, respectively) showed a significant block of the cocaine-induced increase in DA release in the first hour (P<.001) and 5-HT release in both hours of study (P<.001). Cocaine-induced locomotion and stereotypy were blocked simultaneously with the monoamines (P<.001). In subacute studies, DA and 5-HT release returned to baseline while locomotion and stereotypy increased insignificantly above baseline. Thus, (a) these studies were able to tease out pharmacologically the critical differences between presynaptic and postsynaptic responses to drug treatment(s) and these differences may lead to more effective therapies for schizophrenic and/or cocaine psychosis. (b) Taken together with other data, these acute studies suggest that risperidone may possibly act via inhibition of presynaptic autoreceptors to produce the observed increases in accumbens DA and 5-HT release, whereas cocaine may be acting at least in part via serotoninergic modulation of DA postsynaptically. The subacute data suggest that pharmacokinetics may play a role in risperidone's action and that neuroadaptation may play a role in the mechanism of action of cocaine. Finally, the ability of risperidone to block cocaine-induced psychostimulant neurochemistry and behavior during acute studies while diminishing the withdrawal symptoms of cocaine during subacute studies suggests that risperidone may be a viable pharmacotherapy for cocaine psychosis and withdrawal.

Cocaine acts on accumbens monoamines and locomotor behavior via a 5-HT2A/2C receptor mechanism as shown by ketanserin: 24-h follow-up studies.

It is well known that cocaine's psychomotor stimulant properties derive from enhanced monoamines via synaptic transporter/reuptake inhibition and release mechanisms. However, to further understand mechanisms of action for cocaine, which may be receptor-related, ketanserin, a selective 5-HT(2A/2C) antagonist was used to ascertain a possible mediation for 5-HT(2A//2C) receptors in the monoamine and behavioral responses to cocaine. The studies were performed in the freely moving and behaving animal with In Vivo Microvoltammetry. Miniature carbon sensors, BRODERICK PROBE microelectrodes detected dopamine (DA) and serotonin (5-HT) concentrations in Nucleus Accumbens (NAcc) of male, Sprague-Dawley laboratory rats in separate signals and within seconds while at the same time, locomotor behavior was monitored with infrared photobeams. Synaptic release of each monoamine was detected because separate studies showed that the depolarization blocker, gamma-butyrolactone (gamma BL), decreased steady-state values [Pharmacol. Biochem. Behav. 40 (1991) 969]. Acute studies (Day 1) were performed; the animals received single injection of drug(s) in the faradaic behavioral chamber after a stable baseline during habituation behavior was achieved. After completion of the study, the animals were returned to their home cages. Subacute studies (Day 2) were also performed; these took place 24 h later in the faradaic behavioral chamber; same animal control was used and no further drug was administered. Day 2 data were compared to baseline (habituation data) on Day 1. Results showed that (1) Acute administration of Cocaine (10 mg/kg, i.p.) (N=5) increased DA and 5-HT release above baseline (p<0.001) while locomotion was also increased above baseline (p<0.001). (2) In Subacute studies in the cocaine group, when no further drug was administered, DA release decreased (p<0.001) and decreases in 5-HT release also occurred throughout the time course (p<0.05). Locomotor behavior increased above baseline and showed a trend toward statistical significance (p<0.07). (3) Acute administration of Ketanserin/Cocaine (3 mg/kg s.c. and 10 mg/kg i.p., respectively) (N=6) showed that ketanserin antagonized DA and 5-HT release (p<0.001), while locomotion was antagonized as well (p<0.001). (4) In Subacute studies, in the ketanserin/cocaine group, when no further drug was administered, DA decreased (p<0.001), but 5-HT increased (p<0.001), while locomotor activity increased above baseline and a trend toward statistical significance was seen (p<0.07). Additional saline controls were without effect (p>0.05). In summary, Acute studies showed that cocaine produced its psychostimulant responses on monoamines and behavior and ketanserin antagonized these responses, likely via a 5-HT(2A/2C) receptor mediation. Presynaptic and postsynaptic responses were not distinguished, suggesting, in addition, a role for 5-HT-ergic modulation of DA, likely DA(2) postsynaptic modulation Subacute cocaine studies showed that on Day 2, deficiencies in monoamines occurred, reflecting cocaine withdrawal mechanisms neurochemically, while locomotor behavior did not show such dramatic deficiencies. Indeed, behavior increased above baseline. Moreover, ketanserin reversed 5-HT-related and not DA-related cocaine monoamine responses, while locomotion continued to be insignificantly increased above baseline as was seen in the Subacute cocaine group. The data suggest that presynaptic 5-HT(2A/2C) receptor mechanisms may be important during withdrawal from single injection of cocaine.

Clozapine and cocaine effects on dopamine and serotonin release in nucleus accumbens during psychostimulant behavior and withdrawal.

There is an increasing awareness that a psychosis, similar to that of schizophrenic psychosis, can be derived from cocaine addiction. Thus, the prototypical atypical antipsychotic medication, clozapine, a 5-HT(2)/DA(2) antagonist, was studied for its effects on cocaine-induced dopamine (DA) and serotonin (5-HT) release in nucleus accumbens (NAcc) of behaving male Sprague-Dawley laboratory rats with In Vivo Microvoltammetry, while animals' locomotor (forward ambulations), an A(10) behavior, was monitored at the same time with infrared photobeams. Release mechanisms for monoamines were determined by using a depolarization blocker, gamma-butyrolactone (gammaBL). BRODERICK PROBE microelectrodes selectively detected release of DA and 5-HT within seconds and sequentially in A(10) nerve terminals, NAcc. Acute and subacute studies were performed for each treatment group. Acute studies are defined as single injection of drug(s) after a stable baseline of each monoamine and locomotor behavior has been achieved. Subacute studies are defined as 24-h follow-up studies on each monoamine and locomotor behavior, in the same animal at which time, no further drug was administered. Results showed that (1) acute administration of cocaine (10 mg/kg ip) (n=5) significantly increased both DA and 5-HT release above baseline (P<.001) while locomotion was also significantly increased above baseline (P<.001). In subacute studies, DA release decreased significantly below baseline (P<.001) and significant decreases in 5-HT release occurred at the 15-min mark and at each time point during the second part of the hour (P<.05); the maximum decrease in 5-HT was 40% below baseline. Locomotor behavior, on the other hand, increased significantly above baseline (P<.05). (2) Acute administration of clozapine/cocaine (20 and 10 mg/kg ip, respectively; n=6) produced a significant block of the cocaine-induced increase in DA (P<.001) and 5-HT release (P<.001). Cocaine-induced locomotion was blocked simultaneously with each monoamine by clozapine as well (P<.001). In subacute studies, DA release continued to be blocked presumably via clozapine by exhibiting a statistically significant decrease (P<.001), but 5-HT release increased significantly (P<.001), while cocaine-induced locomotor activity also continued to be antagonized by clozapine, i.e., locomotor activity exhibited no difference from baseline (P>.05). In summary, acute studies (a) support previous data from this laboratory and others that cocaine acts as a stimulant on the monoamines, DA and 5-HT and on locomotor behavior as well and (b) show that clozapine, 5-HT(2)/DA(2) antagonist, blocked enhanced DA, 5-HT and psychomotor stimulant behavior induced by cocaine. Subacute studies (a) suggest that withdrawal responses occurred in the cocaine group, based on recorded deficiencies in monoamine neurotransmitters (b) show that withdrawal effects in the cocaine group likely presynaptic, were distinguished from locomotor behavior, classically known to be mediated postsynaptically, and finally, (c) suggest that clozapine, with longer lived pharmacokinetic properties, reversed 5-HT cocaine-related withdrawal effects, but was unable to reverse DA cocaine-related withdrawal responses. Taken together with data from this laboratory, in which the 5-HT(2A/2C) antagonist, ketanserin, affected cocaine neurochemistry in much the same way as did clozapine, a mediation by either separate or combined 5-HT(2A/2C) receptors for these clozapine/cocaine interactions, is suggested. Further studies, designed to tease out the responses of selective 5-HT(2A) and 5-HT(2C) receptor compounds to cocaine and clozapine/cocaine, are underway.

Cocaine Shifts the Estrus Cycle Out of Phase and Caffeine Restores It.

Background: Sex differences in cocaine abuse are well established. Females have a higher sensitivity and thus higher vulnerability to cocaine abuse compared to males. There are many studies showing that sensitivity to cocaine reward varies during the estrus cycle. Methods: Vaginal smears were examined through a DIFF staining kit and viewed through a microscope to determine the estrus cycle stage. Smears were taken immediately before and after cocaine and/or caffeine injections. Furthermore, we suggest a new tool to analyze the estrus cycle by using electrical resistance of the vaginal mucosa. Results: In the present study, we discovered that cocaine directly induced changes in the estrus cycle. Interestingly, caffeine did not affect the estrus cycle and nor did the combination of cocaine and caffeine. We observed that caffeine blocked the cocaine-induced estrus cycle changes using conventional exfoliate cytology. Therefore, caffeine may have neuroprotective properties on the changes induced by cocaine. Conclusion: These phase changes in the estrus cycle may be the underlying cause of sex differences in cocaine addiction that can be blocked by caffeine. Thus, we propose a valuable insight into sex differences in cocaine abuse and reveal a possible treatment with antagonizing the adenosine system.

Caffeine's Attenuation of Cocaine-Induced Dopamine Release by Inhibition of Adenosine.

Background: It is well known that the reinforcing properties of cocaine addiction are caused by the sharp increase of dopamine (DA) in the reward areas of the brain. However, other mechanisms have been speculated to contribute to the increase. Adenosine is one system that is associated with the sleep-wake cycle and is most important in regulating neuronal activity. Thus, more and more evidence is pointing to its involvement in regulating DA release. The current study set out to examine the role of adenosine in cocaine-induced DA release. Methods: Increasing doses of cocaine, caffeine, and their combination, as well as, 8-cyclopentyltheophylline (CPT), an adenosine A1 antagonist (alone and in combination with cocaine) were used to denote a response curve. A novel biosensor, the BRODERICK PROBE® was implanted in the nucleus accumbens to image the drug-induced surge of DA release in vivo, in the freely moving animal in real time. Results: Combinations of cocaine and caffeine were observed to block the increased release of DA moderately after administration of the low dose (2.5 mg/kg cocaine and 12.5 mg/kg caffeine) and dramatically after administration of the high dose (10 mg/kg cocaine and 50 mg/kg caffeine), suggesting neuroprotection. Similarly, CPT and cocaine showed a decreased DA surge when administered in combination. Thus, the low and high dose of a nonselective adenosine antagonist, caffeine, and a moderate dose of a selective adenosine antagonist, CPT, protected against the cocaine-induced DA release. Conclusions: These results show a significant interaction between adenosine and DA release and suggest therapeutic options for cocaine addiction and disorders associated with DA dysfunction.

Neuromolecular Imaging Shows Temporal Synchrony Patterns between Serotonin and Movement within Neuronal Motor Circuits in the Brain.

The present discourse links the electrical and chemical properties of the brain with neurotransmitters and movement behaviors to further elucidate strategies to diagnose and treat brain disease. Neuromolecular imaging (NMI), based on electrochemical principles, is used to detect serotonin in nerve terminals (dorsal and ventral striata) and somatodendrites (ventral tegmentum) of reward/motor mesocorticolimbic and nigrostriatal brain circuits. Neuronal release of serotonin is detected at the same time and in the same animal, freely moving and unrestrained, while open-field behaviors are monitored via infrared photobeams. The purpose is to emphasize the unique ability of NMI and the BRODERICK PROBE® biosensors to empirically image a pattern of temporal synchrony, previously reported, for example, in Aplysia using central pattern generators (CPGs), serotonin and cerebral peptide-2. Temporal synchrony is reviewed within the context of the literature on central pattern generators, neurotransmitters and movement disorders. Specifically, temporal synchrony data are derived from studies on psychostimulant behavior with and without cocaine while at the same time and continuously, serotonin release in motor neurons within basal ganglia, is detected. The results show that temporal synchrony between the neurotransmitter, serotonin and natural movement occurs when the brain is NOT injured via, e.g., trauma, addictive drugs or psychiatric illness. In striking contrast, in the case of serotonin and cocaine-induced psychostimulant behavior, a different form of synchrony and also asynchrony can occur. Thus, the known dysfunctional movement behavior produced by cocaine may well be related to the loss of temporal synchrony, the loss of the ability to match serotonin in brain with motor activity. The empirical study of temporal synchrony patterns in humans and animals may be more relevant to the dynamics of motor circuits and movement behaviors than are studies of static parameters currently relied upon within the realms of science and medicine. There are myriad applications for the use of NMI to discover clinically relevant diagnoses and treatments for brain disease involving the motor system.

Sex-specific brain deficits in auditory processing in an animal model of cocaine-related schizophrenic disorders.

Cocaine is a psychostimulant in the pharmacological class of drugs called Local Anesthetics. Interestingly, cocaine is the only drug in this class that has a chemical formula comprised of a tropane ring and is, moreover, addictive. The correlation between tropane and addiction is well-studied. Another well-studied correlation is that between psychosis induced by cocaine and that psychosis endogenously present in the schizophrenic patient. Indeed, both of these psychoses exhibit much the same behavioral as well as neurochemical properties across species. Therefore, in order to study the link between schizophrenia and cocaine addiction, we used a behavioral paradigm called Acoustic Startle. We used this acoustic startle paradigm in female versus male Sprague-Dawley animals to discriminate possible sex differences in responses to startle. The startle method operates through auditory pathways in brain via a network of sensorimotor gating processes within auditory cortex, cochlear nuclei, inferior and superior colliculi, pontine reticular nuclei, in addition to mesocorticolimbic brain reward and nigrostriatal motor circuitries. This paper is the first to report sex differences to acoustic stimuli in Sprague-Dawley animals (Rattus norvegicus) although such gender responses to acoustic startle have been reported in humans (Swerdlow et al. 1997 [1]). The startle method monitors pre-pulse inhibition (PPI) as a measure of the loss of sensorimotor gating in the brain's neuronal auditory network; auditory deficiencies can lead to sensory overload and subsequently cognitive dysfunction. Cocaine addicts and schizophrenic patients as well as cocaine treated animals are reported to exhibit symptoms of defective PPI (Geyer et al., 2001 [2]). Key findings are: (a) Cocaine significantly reduced PPI in both sexes. (b) Females were significantly more sensitive than males; reduced PPI was greater in females than in males. (c) Physiological saline had no effect on startle in either sex. Thus, the data elucidate gender-specificity to the startle response in animals. Finally, preliminary studies show the effect of cocaine on acoustic startle in tandem with effects on estrous cycle. The data further suggest that hormones may play a role in these sex differences to acoustic startle reported herein.

Real Time Imaging of Biomarkers in the Parkinson's Brain Using Mini-Implantable Biosensors. II. Pharmaceutical Therapy with Bromocriptine.

We used Neuromolecular Imaging (NMI) and trademarked BRODERICK PROBE® mini-implantable biosensors, to selectively and separately detect neurotransmitters in vivo, on line, within seconds in the dorsal striatal brain of the Parkinson's Disease (PD) animal model. We directly compared our results derived from PD to the normal striatal brain of the non-Parkinson's Disease (non-PD) animal. This advanced biotechnology enabled the imaging of dopamine (DA), serotonin (5-HT), homovanillic acid (HVA) a metabolite of DA, L-tryptophan (L-TP) a precursor to 5-HT and peptides, dynorphin A 1-17 (Dyn A) and somatostatin (somatostatin releasing inhibitory factor) (SRIF). Each neurotransmitter and neurochemical was imaged at a signature electroactive oxidation/half-wave potential in dorsal striatum of the PD as compared with the non-PD animal. Both endogenous and bromocriptine-treated neurochemical profiles in PD and non-PD were imaged using the same experimental paradigm and detection sensitivities. Results showed that we have found significant neurotransmitter peptide biomarkers in the dorsal striatal brain of endogenous and bromocriptine-treated PD animals. The peptide biomarkers were not imaged in dorsal striatal brain of non-PD animals, either endogenously or bromocriptine-treated. These findings provide new pharmacotherapeutic strategies for PD patients. Thus, our findings are highly applicable to the clinical treatment of PD.

Novel research translates to clinical cases of schizophrenic and cocaine psychosis.

Pharmacotherapies for schizophrenic and cocaine psychoses are complex but similar because of similarities in their brain neurochemistry and behavioral outcomes. Their neurochemical neuronal mechanisms of action, as shown in preclinical and clinical studies, involve primarily dopaminergic dysfunction and, secondarily, neuroadaptive effects that seem to involve central serotonergic function. Behavioral outcomes of both disorders include hyperactivity and antipsychotic medications can ameliorate psychotic symptoms. Patients with both disorders often arrive at emergency departments and present floridly psychotic with a predominance of positive symptoms, often prompting physicians to select a typical antipsychotic medication such as haloperidol. While this has become conventional wisdom, we believe that to use an atypical antipsychotic medication, such as risperidone, in the treatment of both psychoses is quite rational for long-term management of both positive and negative symptoms. Also, controlled clinical studies have shown that risperidone, an atypical antipsychotic medication, is successful in the treatment of cocaine dependence and withdrawal (Smelson et al 1997, 2002; Grabowski et al 2000). Furthermore, the availability and effectiveness of long-acting risperidone in injectable form opens new possibilities for the long-term management of both disorders. In this paper, we present data which show that the use of risperidone is plausible for effective pharmacotherapy of schizophrenic and cocaine psychoses.

Central monoamine dysfunction in diabetes: psychotherapeutic implications; electroanalysis by voltammetry

Se examinaron en ratas, mediante voltametría, los efectos de la diabetes inducida por estreptozotocina o de la hiperglucemia, sobre la liberción de dopamina y serotonina en el estriado in vivo. En el estado diabético agudo se observó un incremento del 67% en la liberación de dopamina, mientras que el incremento observado en la diabetes crónica fue menor (19%). En las ratas a los 3 días de inducida la diabetes la señal electroquímica correspondiente a serotonina aumentó en un 62%, efecto que desapareció en la diabetes crónica. Luego de la inyección de L-triptófano en ratas normales, se detectó una disminución del 45% en la dopamina estriatal liberada y un aumento del 25% en la liberación de serotonina. Este incremento fue máximo a los 90 min más. Los animales crónicamente diabéticos mstraron una disminución significativa en la liberación de dopamina y serotonina estriatal luego de inyectar triptófano. Los efectos de la hiperflucemia en ratas no diabéticas fueron una disminución (52%) de la liberación de dopamina, y un incremento (304%) de la liberación de serotonina. Estos cambios indican que el estado diabético no tratado se asocia con una disminución progresiva de la liberación de neurotrasmisor, y que las modificaciones emocionales observadas en la disponibilidad de dopamina y serotonina en sinapsis centrales

Central monoamine dysfunction in diabetes: psychotherapeutic implications; electroanalysis by voltammetry

Se examinaron en ratas, mediante voltametría, los efectos de la diabetes inducida por estreptozotocina o de la hiperglucemia, sobre la liberción de dopamina y serotonina en el estriado in vivo. En el estado diabético agudo se observó un incremento del 67% en la liberación de dopamina, mientras que el incremento observado en la diabetes crónica fue menor (19%). En las ratas a los 3 días de inducida la diabetes la señal electroquímica correspondiente a serotonina aumentó en un 62%, efecto que desapareció en la diabetes crónica. Luego de la inyección de L-triptófano en ratas normales, se detectó una disminución del 45% en la dopamina estriatal liberada y un aumento del 25% en la liberación de serotonina. Este incremento fue máximo a los 90 min más. Los animales crónicamente diabéticos mstraron una disminución significativa en la liberación de dopamina y serotonina estriatal luego de inyectar triptófano. Los efectos de la hiperflucemia en ratas no diabéticas fueron una disminución (52%) de la liberación de dopamina, y un incremento (304%) de la liberación de serotonina. Estos cambios indican que el estado diabético no tratado se asocia con una disminución progresiva de la liberación de neurotrasmisor, y que las modificaciones emocionales observadas en la disponibilidad de dopamina y serotonina en sinapsis centrales (AU)