Three-dimensional mapping of norepinephrine and serotonin in human thalamus.

Detailed quantitative information on catecholamines and 5-hydroxytryptamine (serotonin) in the human thalamus is much needed because of increasing interest in norepinephrine and serotonin as modulators of thalamic behavioral state control and overall information processing. This study provides three-dimensional distribution patterns of these monoamines in postmortem thalami from 13 normal subjects (no known neurological or psychiatric histories). The patterns come from a relatively fine-grained grid mapping procedure on successive coronal sections. Samples were analyzed by high performance liquid chromatography with electrochemical detection. The highest endogenous concentrations of norepinephrine are found in a ventromedial core that includes a number of the medial and intralaminar sub-nuclei but extends only slightly into the sensory regions of the lateral tier. The posterior portion of the thalamus, the pulvinar, contains low levels of norepinephrine. The distribution of 5-hydroxytryptamine is quite similar to that of norepinephrine in the rostral two-thirds of thalamus; however, in the pulvinar region, levels of serotonin are considerably increased and differ markedly between individual thalami. The study provides the first definitive mapping of serotonin levels in human thalamus. Consistent with many animal studies, there is no evidence for major dopaminergic innervation of human thalamus. By emphasizing the pattern distribution of the monoamines rather than the absolute values, it can be shown that the ambiguities of postmortem degradation frequently associated with biochemical assays are largely avoided. The terminal field distribution of norepinephrine is an essentially constant neurochemical signature in all thalami examined. The utility of the biochemical grid mapping procedure may be especially significant in terms of matching with data from functional neuroimaging techniques.

In vivo dynamics of norepinephrine release-reuptake in multiple terminal field regions of rat brain.

Electrical stimulation of the ascending dorsal tegmental bundle of the locus ceruleus was used to elicit controlled release of norepinephrine. Real-time in vivo monitoring in the brains of urethane-anesthetized rats was observed with high speed chronocoulometry at rapidly responding carbon fiber electrodes. Using modeling similar to that developed for dopamine release, the electrochemical signals were characterized as the balance between norepinephrine release per electrical stimulation pulse and apparent Michaelis-Menten reuptake parameters. Stimulation produced simultaneous overflow release at all terminal fields examined. The release and reuptake characteristics varied considerably in different regions. If the parameters are normalized to endogenous concentration in the terminal fields, release but not reuptake correlates with innervation density in several regions. Stimulated release results in norepinephrine overflow and transport in most brain regions with half-lives of 1-3 s and overflow distances of 25-50 microns at most. A surprising exception occurs in the upper layers of cortex (cingulate and sensory) where half-lives may be in the 10s of seconds and spatial reach may be up to 100 microns. The uptake in the outer cortical layers appears to be minimal and comparable with only nonspecific reuptake.

Neuroleptic treatment is an unlikely cause of elevated dopamine in thalamus of schizophrenic subjects.

Previous studies have indicated a marked increase in the dopamine/norepinephrine ratio in thalami of schizophrenic patients compared with those of control subjects. Since these results all came from patients who were receiving neuroleptic drugs, the possibility exists that the increased dopamine concentrations are an effect of medication. To address this question, similar analyses were done on thalami from Huntington's Disease patients who had received neuroleptic treatment. The results showed no differences between the thalami of Huntington's Disease patients and controls, strongly suggesting that chronic treatment with neuroleptic drugs does not result in an increase of endogenous dopamine in the thalami of human subjects.

Mechanical slicing of frozen brain tissue: a reappraisal of catecholamine loss.

Catecholamine levels in mechanically sliced frozen bovine brain tissue were compared with those found in hand-cut, unfrozen tissue. Hemispheric content of catecholamines in small regions of cortex, caudate, nucleus accumbens, thalamus, and hippocampus in bovine brain was no different from that seen in the homologous area of the contralateral hemisphere, despite selective slicing of opposing hemispheres with the two techniques. Brodmann area 25 in human brain (previously suspected to be highly vulnerable to degradative loss of catecholamines by mechanical slicing) is shown to be a restricted area of dense catecholamine concentration when compared to surrounding cortical tissue. The usage of Brodmann area 25 for technique comparisons without precise control of cortical delineation is questioned.

Elevated thalamic dopamine: possible link to sensory dysfunctions in schizophrenia.

Sensory-processing dysfunctions, deficit states, and the combinations of seemingly disparate behavioral symptoms of schizophrenia are addressed with regard to a common thread--the possibility of dysfunctional processing in the thalamus. Recent views of the connectional neuroanatomy and electrical activity of thalamus are examined. A hypothesis is presented in which disturbances in the timing and phasic neuronal activity of the thalamus and, especially, its connections with other brain regions may result in many of the behavioral manifestations of schizophrenia. It is suggested that neurotransmitter or other chemical imbalances might produce such thalamic disturbances. Experimental findings of enhanced dopamine content in the thalami of schizophrenic patients are reported. Several varieties of distributional patterns of this elevated dopamine are shown and evaluated.

Effect of CO2 on a brain extracellular space marker and evidence of its neuronal modulation.

Increases in inspired CO2 consistently altered the local concentration of the brain extracellular space marker alpha-naphthalene sulfonate (alpha-NS) as measured with ion-selective micropipettes in the rat thalamus. Stereotaxic injection of lidocaine in the region of the locus coeruleus attenuated this effect of CO2, and amitriptyline, a tricyclic anti-depressant and amine reuptake inhibitor, potentiated the effect. These results suggest that metabolic demand, as mimicked here by the addition of CO2, alters the fluid environment of the brain and central noradrenergic mechanisms may modulate this response.

Simultaneous voltammetric and chemical monitoring of dopamine release in situ.

The validity of voltammetric monitoring of stimulated dopamine (DA) release was tested by simultaneous chemical assay of the extracellular fluid region next to the tip of a selective Nafion-coated graphite voltammetric electrode. Micro alumina probes were positioned next to the recording electrode tip to adsorb released catecholamines. The catecholamines were desorbed into microvolumes of acid and analyzed by very sensitive HPLC assays to provide unequivocal chemical verification of the electrochemical signals. The results fully confirm that selective voltammetric electrodes can detect released DA and that the concentrations determined voltammetrically or chemically agree well.

Nafion-coated electrodes with high selectivity for CNS electrochemistry.

A major improvement in the selectivity of small graphite electrodes used for in vivo electrochemistry is described. The electrodes are coated with Nafion, a perfluorosulfonated polymer. This coating is practically impermeable to ascorbic acid and anionic biogenic amine metabolites and only slightly responsive to neutral metabolites. Thus it becomes selective for the cationic primary neurotransmitters, dopamine, norepinephrine and 5-hydroxytryptamine. Responses of Nafion-coated and untreated electrodes in vivo are compared.

Regional distribution of ascorbate in human brain.

A comprehensive mapping of ascorbate distribution in human brain was carried out by liquid chromatography analysis. The data agree with earlier literature values where comparable and provide new information on several brain regions, including a detailed distribution in the thalamus. While ascorbate concentrations tend to be high in regions rich in catecholamines, there is no real correlation between the two.

Selective attention dysfunctions in adult rats neonatally treated with 6-hydoxydopamine.

Adult rats treated intracisternally with 6-hydroxydopamine during the perinatal period were trained on a black-white discrimination. Alterations in cue shape and cue location failed to selectively distinguish differences in response patterns for the treated animals. However, when irrelevant stimuli were added to the discriminative array, the 6-hydroxydopamine group evidenced marked impairment of performance throughout the period of distraction.