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Biomedical subjects

C R Freed

Publications and source records attributed to C R Freed.

At least 73 records · Page 4Linked to original sources
PubMed1987

In vivo electrochemical recording of acetaminophen in non human primate brain.

In vivo electrochemical recordings were obtained from the caudate nucleus of three young adult pigtail monkeys (M. nemestrina) following the oral administration of acetaminophen (APAP) (75 mg/kg). Using linear sweep voltammetry, electrodes in the right and left caudate were scanned alternately every five minutes. The electrochemical peak resulting from the APAP was monitored for at least 140 minutes following drug administration. Maximum APAP levels were detected in the monkey caudate 40 minutes following drug administration. Both right and left caudate displayed an identical time course, with oxidation potentials (Eox) similar on both sides of the brain. Blood samples were collected from one monkey by means of an intravenous catheter. Samples were obtained at approximately 5-minute intervals and over a period of 140 minutes following oral administration of APAP. Concentration of APAP in serum peaked 25 minutes after administration. In this animal the maximum electrochemical peak height was detected 40 minutes following APAP administration. These findings demonstrate the ability to measure APAP in the caudate nucleus of awake monkeys by means of electrochemical detection. This method may be useful for calibrating electrochemical electrodes in vivo, and it also provides a model system for studying drug kinetics in the brain.

Acetaminophen↗
PubMed1987

Wire electrodes for chronic single unit recording of dopamine cells in substantia nigra pars compacta of awake rats.

Stainless steel wire electrodes of varying sizes and coated with different insulating materials were tested in order to find a flexible fine wire which would allow the recording of small cells in deep structures of the brain in unrestrained, awake rats. Our main interest is to record from cells of substantia nigra pars compacta during locomotion. We found that stress relieved 18 micron stainless steel wire doubly insulated with lacquer and Parylene C provided the impedance, physical size, and flexibility needed to record single units during intense motor behavior.

Animals↗
PubMed1986

Factors affecting in vivo electrochemistry: electrode-tissue interaction and the ascorbate amplification effect.

While in vivo electrochemistry has been shown to be useful for discovering new neurophysiological phenomena, there is still considerable controversy about the identity of the compounds being measured and the concentration of those compounds in extracellular fluid in brain. We have found that carbon paste electrodes undergo changes in sensitivity and specificity for dopamine and other compounds after being implanted in brain. We have also examined the effect of ascorbate on the selective enhancement of catecholamine peaks to provide an explanation for the apparently very high concentrations of dopamine measured in the extracellular fluid space. After temporary brain implantation (20 min), carbon paste electrodes tested in vitro showed increased sensitivity and lower oxidation potentials for dopamine, norepinephrine and serotonin. These brain-treated electrodes also detected 3,4-dihydroxyphenylacetic acid (DOPAC) as a distinct peak at +0.16 V, although the electrode sensitivity for DOPAC was some 25 times lower than that for dopamine. Brain treatment did not alter electrode sensitivity or oxidation potential for 5-HIAA. The oxidation current for ascorbic acid when processed as the semiderivative showed no distinct peak in the potential range -0.2 to +0.4V for either untreated or brain-treated electrodes. However ascorbic acid amplified the electrochemical peaks of catechols in direct proportion to the ratio of the concentration of ascorbate to the concentration of the catechol. In the physiologic concentration range of 300 microM ascorbate, the electrochemical signal for 1 microM dopamine was amplified 4250%. While ascorbate amplification improves detectability of dopamine and norepinephrine, it also introduces ambiguity since changing catechol concentrations cannot be distinguished from changing ascorbate concentrations.

Animals↗
PubMed1986

Mobilization of storage pool dopamine and late ipsilateral augmentation of striatal dopamine synthesis in the trained circling rat.

Rats were infused intraventricularly with [3H]tyrosine over a 20-min period during various times while circling. 3,4-Dihydroxyphenylethylamine (dopamine) and dihydroxyphenylacetic acid (DOPAC) levels were measured using HPLC with electrochemical detection and fractions were collected for tritium monitoring. During the first 20 min of circling, the specific activity of dopamine was increased by 290% in striatum contralateral to the circling direction whereas DOPAC specific activity was increased 50% on the same side. This differential change in relative specific activity suggests that unlabeled storage pool dopamine was mobilized to DOPAC during circling. Synthesis of dopamine and DOPAC in contralateral striatum returned to baseline levels as turning slowed (50-70 min). When turning ceased, there was an increase in ipsilateral striatal dopamine synthesis during the 20-min period following circling. We hypothesize that this ipsilateral increase represents either a "stop" signal following circling or a release of inhibition of ipsilateral nigral neurons.

3,4-Dihydroxyphenylacetic Acid↗
PubMed1985

Brain serotonin and blood pressure regulation: studies using in vivo electrochemistry and direct tissue assay.

Hypotensive responses to tryptophan and 5-hydroxytryptophan infusions were studied in normotensive male Sprague-Dawley rats. Results showed that 5-hydroxytryptophan but not tryptophan lowered pressure in a dose dependent way in direct relation to the production of brain serotonin and 5-HIAA. Intrinsic release of serotonin from brain was also studied during periods of induced hypertension and hypotension. Brain monoamine responses to blood pressure changes induced by intravenous phenylephrine and nitroprusside were measured in dorsal raphe nucleus and nucleus tractus solitarius by in vivo electrochemistry. Results showed that 5-HIAA was increased during drug induced hypertension and during reflex hypertension which followed a period of hypotension. These changes were blocked by sinoaortic denervation indicating that these central serotonergic neurons are responding to increased pressure sensed by baroreceptors. Therefore, serotonin has a role in blood pressure regulation as a pharmacologic agent and as a neurotransmitter in homeostatic control of pressure.

5-Hydroxytryptophan↗
PubMed1985

Regional brain dopamine metabolism: a marker for the speed, direction, and posture of moving animals.

Brain dopamine is necessary for normal movement. To determine whether there is a precise relation between the intensity of movement and changes in brain dopamine metabolism, the investigators ran rats on straight and circular treadmills at different speeds and with different body postures. Concentrations of dopamine and its metabolite 3,4-dihydroxyphenylacetic acid increased in the caudate and accumbens nuclei in direct relation to the speed and angular posture of the animals. Dopamine metabolism in the nucleus accumbens was more strongly linked to the speed and direction of movement, while in the caudate nucleus dopamine and 3,4-dihydroxyphenylacetic acid were affected most by posture and direction.

3,4-Dihydroxyphenylacetic Acid↗
PubMed1984

Caudate electrochemical response following amphetamine administration in pigtail monkeys.

In vivo electrochemical and heart rate (HR) recordings following amphetamine (AMPH) (0.8 mg/kg) and saline administration were made from caudate in four young adult pigtail (M. nemestrina) monkeys using linear sweep voltammetry. One hour following drug injection, two familiar humans served as test stimuli, and were visually exposed to the animals for 15-minute epochs each. One was threatening to the animals, and one was not. AMPH produced a significant increase in height of the electrochemical peak thought to represent oxidation of dopamine and its metabolites. Heart rate (HR) decreased during the time the peak height was increasing. HR and peak height increased during presentation of both humans under both AMPH and saline conditions. However, peak height increase under AMPH, but not saline, conditions discriminated the negative from neutral stimulus. The findings demonstrate that AMPH administration induces a significant increase in the height of a major electroactive peak in the caudate nucleus of pigtail monkeys, and further that such amphetamine-induced increases can be manipulated by altering the affective and/or emotional state of the animal.

Animals↗
PubMed1984

Altered serotonin and norepinephrine metabolism in rat dorsal raphe nucleus after drug-induced hypertension.

The effect of drug-induced hypertension on neurotransmitter release from dorsal raphe nucleus was studied by in vivo electrochemical electrodes in urethane anesthetized male Sprague-Dawley rats. Carbon paste electrodes were stereotaxically placed into dorsal raphe nucleus and neurotransmitter release was estimated electrochemically. Blood pressure was recorded from a femoral arterial catheter. Voltammograms taken from dorsal raphe nucleus showed two distinct peaks corresponding to norepinephrine and 5-hydroxyindole acetic acid (5-HIAA). After basal blood pressure and neurotransmitter release were monitored for 30 min, blood pressure was raised 50 mmHg by continuous intravenous infusion of L-phenylephrine hydrochloride. Drug infusion was discontinued after 50 min, but blood pressure and neurotransmitter release were measured for an additional 2 hr. Results showed that the 5-HIAA response increased immediately after the initiation of hypertension and remained elevated. By contrast, norepinephrine release initially decreased, then returned to the basal level and then rose in parallel with 5-HIAA to a level above baseline as drug-induced hypertension was discontinued. The same experimental protocol was used to study the electrochemical response to drug-induced hypotension. Blood pressure was lowered 20 mmHg by intravenous infusion of sodium nitroprusside dihydrate. During hypotension, no changes were seen in either transmitter response. However, as reflex hypertension appeared following discontinuation of the sodium nitroprusside infusion, the 5-HIAA response increased and the norepinephrine response decreased. These results show that drug-induced and reflex hypertension reduce norepinephrine release and increase serotonin turnover in dorsal raphe nucleus in anesthetized normotensive rats. These reciprocal changes appear to be a part of the neural response to hypertension.

Animals↗
PubMed1984

Asymmetric dopamine and serotonin metabolism in nigrostriatal and limbic structures of the trained circling rat.

The trained circling rat model was used to investigate dopamine and serotonin metabolism in extrapyramidal and limbic structures during turning behavior. We have previously reported that dopamine turnover is increased during circling in the caudate contralateral to the circling direction in this behavioral model. We have now studied changes in dopamine and serotonin turnover in nucleus accumbens, substantia nigra and amygdala. As in the caudate, dopamine production in nucleus accumbens was selectively increased on the contralateral side after 20 min of circling. By contrast, dopamine turnover in substantia nigra exhibited a relative decline on the contralateral side. Dopamine synthesis in the amygdala was not affected by circling. Selective changes in serotonin metabolism were also seen in these brain regions. In caudate and accumbens, serotonin turnover was unaffected by circling. However, both substantia nigra and amygdala showed significant, progressive increases in serotonin metabolism in the contralateral side after 20 and 70 min of circling. These results show that extrapyramidal and limbic dopamine and serotonin metabolism are involved in turning behavior of normal animals. Multiple transmitters of the nigrostriatal pathway and the limbic system appear to interact to modulate voluntary circling behavior.

3,4-Dihydroxyphenylacetic Acid↗
PubMed1984

Reversal of amphetamine-induced circling preference in trained circling rats.

Others have shown that amphetamine given to normal rats causes turning in a particular, preferred direction in most animals. We have studied the effect of training on amphetamine-induced behavioral and biochemical asymmetries in male Sprague-Dawley rats. Water deprived animals were trained to circle either in the same or opposite direction to their intrinsic bias using a sucrose water reward. Acquisition of the learned circling behavior was independent of turning preference and all animals were able to make the operant association. After training, animals given amphetamine turned in the trained direction regardless of their previous circling preference. Amphetamine-induced circling also led to increased dopamine concentrations in caudate contralateral to the trained circling direction. Therefore, intrinsic striatal lateralization is not resistant to behavioral modification and both the behavioral and biochemical asymmetries can be reversed by circling training.

3,4-Dihydroxyphenylacetic Acid↗
PubMed1984

Measurement of serotonin turnover rate in rat dorsal raphe nucleus by in vivo electrochemistry.

5-Hydroxytryptamine (5-HT; serotonin) turnover rate in dorsal raphe nucleus of the urethane-anesthetized rat was estimated by using the in vivo electrochemical detector to measure the decay of extraneuronal 5-hydroxyindole acetic acid (5-HIAA) after monoamine oxidase inhibition. Carbon paste electrodes were scanned by semiderivative voltammetry and revealed two peaks: one at +0.15 V and the other at +0.25 V. The higher potential peak is composed primarily of the 5-HT metabolite 5-HIAA. After administration of pargyline, 75 mg/kg i.p., this peak declined exponentially. Regression analysis of these data by an exponential decay model yielded the fractional rate constant 0.82 +/- 0.06 h-1 (mean +/- SEM). This rate constant of 5-HIAA disappearance measured by in vivo electrochemistry is identical to the rate constant found by others measuring 5-HIAA disappearance by direct tissue assay methods. In animals not treated with pargyline, tissue 5-HIAA concentrations in the dorsal raphe nucleus were measured by HPLC with electrochemical detection. The average 5-HT turnover rate calculated as the product of the fractional rate constant and steady-state tissue 5-HIAA concentration was 12.6 nmol/g/h. These results demonstrate that electrochemical detection of extraneuronal 5-HIAA combined with monoamine oxidase inhibition can be used to measure neurotransmitter turnover in vivo in a discrete brain region.

Animals↗
PubMed1984

Unilateral activation of caudate tyrosine hydroxylase during voluntary circling behavior.

We trained rats to circle for a sucrose water reward and found that this behavior is associated with a unilateral increase in the activity of caudate tyrosine hydroxylase, the rate-limiting enzyme for dopamine synthesis. The increase in tyrosine hydroxylase activity occurs in caudate contralateral to the circling direction and the change is transient, increasing during the first 20 min of circling but then plateauing and falling as turning slows. Enhanced synthetic capacity is followed by increases in the contents of dopamine and dihydroxyphenylacetic acid in the contralateral caudate nucleus. These observations are the first evidence for specific activation of a neurotransmitter synthetic enzyme during voluntary motor behavior.

Animals↗
PubMed1983

In vivo electrochemical detection of extraneuronal 5-hydroxyindole acetic acid and norepinephrine in the dorsal raphe nucleus of urethane-anesthetized rats.

In vivo electrochemical detection of endogenous neurotransmitters was done in the dorsal raphe nucleus of urethane-anesthetized male Sprague-Dawley rats. Stereotaxically implanted carbon paste electrodes were scanned using a cyclic voltammetry amplifier with semiderivative signal processing over the potential range--0.2 to +0.5 V at the rate of 10 mV/s. Two distinct peaks were observed at +0.15 V (peak 1) and +0.25 V (peak 2), respectively. Peak identification was assessed by comparing the oxidation potential observed in vivo with those observed in in vitro experiments using pure catecholamines, indoleamines and their metabolites as well as ascorbic acid. Further characterization of in vivo peaks was done by observing changes in electrochemical peaks as well as tissue neurotransmitter concentrations after pharmacological manipulations. p-Chlorophenylalanine, m-hydroxybenzylhydrazine (NSD-1015), pargyline, alpha-methyl-p-tyrosine and fusaric acid were administered in an effort to block catecholamine or serotonin synthesis or degradation. Results of these experiments revealed that peak 1 primarily represents extracellular norepinephrine, while peak 2 is primarily produced by extracellular 5-hydroxyindoleacetic acid (5-HIAA).

Animals↗
PubMed1982

The trained circling rat: a model for inducing unilateral caudate dopamine metabolism.

Despite considerable knowledge about brain dopamine in drug-induced and pathological states, the dynamic relationship of dopamine to voluntary motor behaviour is not established. In an effort to produce selective, lateralized changes in brain dopamine metabolism associated with movement, we have now developed an animal model in which normal rats were trained to circle for a sucrose/water reward. Turning direction was randomly assigned. The effect of circling on dopamine metabolism was studied by killing animals during the reinforcement period and measuring caudate dopamine and dihydroxyphenylacetic acid (DOPAC) concentrations in both sides of brain. Prior to turning, caudate dopamine and DOPAC levels were the same bilaterally. By contrast, animals killed after 20 min of circling showed a 67% increase in dopamine and a 46% increase in DOPAC concentrations in the caudate contralateral to the turning direction while the ipsilateral caudate showed no concentration changes. Such a rapid, massive unilateral increase in caudate dopamine metabolism suggests that dopamine neurones in contralateral caudate are selectively activated during circling behaviour. This model should be useful for further study of the relationships between neurotransmitter activity, regional brain physiology and voluntary movement, without the need for drugs or brain lesions.

3,4-Dihydroxyphenylacetic Acid↗