John D. Lane

Self-administration of methionine enkephalin into the nucleus accumbens

Microinfusions of the endogenous opiate neurohumor, methionine enkephalin, into the nucleus accumbens initiated a reinforcing stimulus in a dose-related manner. The reinforcing nature of this intracranial self-administration was evaluated with intermittent schedules of reinforcement and a two-lever discrimination procedure. Opiate receptors are likely responsible for the initiation of this reinforcing stimulus since naloxone effectively blocked self-administration. These data suggest the mediation of opiate reinforcement through interactions with opiate receptors in brain regions outside the ventral tegmental area, questioning the current dopamine hypothesis for the initiation of these reinforcement processes.

Effects of 6-OHDA lesions of the central medial nucleus accumbens on rat intravenous morphine self-administration

The function of dopaminergic innervations of the central medial nucleus accumbens in the processes maintaining intravenous morphine self-administration was assessed by lesioning with 6-OHDA and comparing drug intake with sham-vehicle treated littermates. Localized bilateral lesions of this structure resulted in significant increases in morphine intake shifting the dose-effect relationship to the right with twice the dose necessary to maintain prelesion rates of self-administration. Content of dopamine and dihydroxyphenylacetic acid was decreased in the nucleus accumbens after the lesion, but unchanged in the adjacent pyriform cortex and anterior caudate nucleus-putamen, while serotonin was significantly decreased in the pyriform cortex. High affinity uptake measurements also suggested nucleus accumbens dopaminergic and pyriform cortex serotonergic innervations to be affected by the lesion. The shift to the right in the dose effect relationship after the lesion suggests these neuronal systems to be excitatory to the processes mediating self-administration.

Brain neurotransmitter turnover correlated with morphine-seeking behavior of rats.

Neurochemical substrates of intravenous opiate self-administration were investigated in rats using littermate controls for vehicle and passive morphine infusion. The rates of turnover of the putative neurotransmitters, dopamine, norepinephrine, serotonin, gamma-aminobutyric acid, aspartate and glutamate were concurrently measured in eleven brain regions of rats intravenously self-administering morphine and yoked-morphine or yoked-vehicle infused littermates. The passive infusion of morphine resulted in significant changes in the rates of turnover of the biogenic monoamine and amino acid neurotransmitters in six brain regions with the caudate nucleus-putamen-globus pallidus showing the most changes. The contingent infusion of morphine resulted in changes in utilization rates that were generally greater in both magnitude and number than the effects of the drug itself. Twenty-nine significant changes were observed in the self-administering group with most changes occurring in limbic structures. The neurotransmitter turnover rate changes resulting from contingent administration suggest that the drug administration environment is an important factor that should be considered in studies of interactions between drugs and neuronal systems.

Turnover Rates of Amino Acid Neurotransmitters in Regions of Rat Cerebellum

Abstract: The turnover rates of aspartate, γ‐aminobutyric acid (GABA), glutamate, glutamine, alanine, serine, and glycine were measured in five regions of rat cerebellum. Turnover rates of the putative neurotransmitters (aspartate, glutamate, and GABA) were 2–20‐fold higher than those of alanine and serine, and generally consistent with the proposed neurotransmitter functions for these amino acids. However, glutamate turnover was high and similar in magnitude in the deep nuclei and granule layer, suggesting possible release, not only from parallel fibers, but from mossy fibers as well. The differential distribution of turnover rates for GABA supports its neuronal release by Purkinje, stellate, basket, and Golgi cells, whereas aspartate may be released by both climbing and mossy fibers. The distribution of glycine turnover rates is consistent with release from Golgi cells, whereas alanine may be released from granule cell parallel fibers. Turnover rates measured in two other motor areas, the striatum and motor cortex, indicated that utilization of these amino acid neurotransmitters is differentially distributed in brain motor regions. The data indicate that turnover rate measurements may be useful in identifying neurotransmitter function where content measurements alone are insufficient.

Biogenic monoamine turnover in discrete rat brain regions is correlated with conditioned emotional response and its conditioning history

The content and turnover of dopamine, norepinephrine and 5-hydroxytryptamine (serotonin), and the content of their respective major metabolites were evaluated in 19 discrete brain areas of rats exposed to conditioned emotional response (CER), and in control groups which received either equivalent yoked shock (shock only) or compound stimulus presentation (tone only). On test day, CER animals suppressed responding and exhibited forms of emotional behavior after presentation of the conditioned stimulus (CS); while shock only and tone only control groups, and CER animals which received an acute dose of diazepam prior to testing, did not suppress. Few changes were observed in content of the biogenic amines or their metabolites, suggesting that the behavioral manipulations were acting within normal physiological limits. On the other hand, numerous changes were observed in the utilization of the 3 biogenic monoamines, which were correlated with the conditioning-anxiety (comparisons of CER vs shock only) and the shock history (comparison of shock only vs tone only). These observations are consistent with putative neural pathways in the frontal cortex, septum, nucleus accumbens, amygdala, striatum, hippocampus and brain stem (which utilize specific monoamines), and with discrete brain areas which have been implicated in classical conditioning and CER-related phenomena. These observations suggest roles for biogenic monoamines in mediating or responding to the classical conditioning and emotional components of the paradigm.

Use of a single compartment LCEC cell in the determinations of biogenic amine content and turnover.

Content and specific radioactivity of the biogenic monoamines and content of their precursors and metabolites were simultaneously determined in CNS tissue extracts with a high pressure liquid chromatography system (HPLC). The content of dopamine, norepinephrine, serotonin. 5-hydroxyindoleacetic acid, homovanillic acid, 3,4-dihydroxyphenylacetic acid and 4-hydroxy-3-methoxyphenylethyleneglycol and the turnover rate of dopamine, norepinephrine and serotonin were measured in discrete rat brain regions using a one compartment electrochemical detector cell coupled to a C18-reverse phase HPLC column. The small fluid volume dead space of the cell allows the direct and precise collection of individual peaks for determining specific radioactivities. This method is especially suitable for central nervous system tissue samples from 8-20 mg wet weight and the sensitivity of the system in its routine configuration is approximately 2 pmol. This method for determinations of turnover is appropriate for investigations of animals in sensitive behavioral paradigms.U

Limbic acetylcholine turnover rates correlated with rat morphine-seeking behaviors

Acetylcholine (ACh) turnover rates were measured in fourteen brain regions of rats intravenously self-administering morphine and in yoked-morphine and yoked-vehicle infused littermates to identify cholinergic neuronal pathways potentially involved in opiate reinforcement processes. Rats receiving chronic passive administration of morphine had increased ACh turnover rates in the frontal cortex and diagonal band and decreased rates in the medial septum. The significant changes in animals self-administering the drug were prominent in limbic regions with increases in the frontal cortex and decreases in the pyriform cortex, nucleus accumbens, amygdala and ventral tegmental area. Some components of opiate reinforcement may be mediated by increases in the activity of cholinergic ventral pallidal and diagonal band fibers innervating the frontal cortex and by decreases in activity of cholinergic fibers innervating the ventral tegmental area. These data and turnover rates for dopamine, norepinephrine, serotonin, aspartate, glutamate and gamma-aminobutyric acid previously determined in similarly treated animals are consistent with two neuronal circuits that may be involved in opiate seeking behaviors and opiate reinforcement processes.

Limbic muscarinic cholinergic and benzodiazepine receptor changes with chronic intravenous morphine and self-administration

Muscarinic cholinergic and benzodiazepine receptor affinities and densities were evaluated in membranes from seven brain regions of rats intravenously self-administering morphine and in littermates receiving yoked-morphine or yoked vehicle infusions to identify neuronal systems potentially involved in mediating opiate reinforcement processes. Passive morphine infusion resulted in increases in muscarinic cholinergic receptor densities in the pyriform cortex and in decreases in the cingulate cortex while benzodiazepine receptor densities were decreased in both the hippocampal formation and entorhinal-subicular cortex compared to littermates receiving passive infusions of vehicle. Morphine self-administration resulted in decreased muscarinic cholinergic receptor densities in the frontal and entorhinal-subicular cortices and increases in the amygdaloid complex compared to littermates receiving yoked passive drug. These data are in agreement with acetylcholine turnover rate measurements in these animals and support the proposed role of cholinergic innervations of the frontal and entorhinal-subicular cortices and amygdaloid complex in opiate reinforcement processes.

Changes in biogenic amine and benzodiazepine receptors correlated with conditioned emotional response and its reversal by diazepam

Groups of littermate rats were trained to respond for food reinforcement on a variable interval one-min (VI 1) schedule, after which they were classically conditioned to associate a conditioned stimulus (CS) with footshock (conditioned emotional response; conditioned suppression; CER). Two control groups received yoked footshock (no CS) or the visual-auditory stimulus only (no footshock). On test day, a group of the CER conditioned animals received injections of either vehicle or diazepam prior to exposure to the VI 1 food-reinforced schedule. After 30 min of the VI 1 schedule, the CS was presented continuously for 15 min, after which the animals were decapitated, the brains removed, membranes prepared and in vitro receptor binding evaluated. During the CS, the CER animals suppressed responding and exhibited conditioned fear (emotional) behavior, while the control groups, and animals given acute diazepam, maintained normal responding. [3H]Diazepam binding was reduced in the CER animals, yet acute benzodiazepine administration did not effect this binding. [3H]QNB binding was reduced by CER and increased by diazepam administration. Adrenergic, serotonergic and dopaminergic systems were also evaluated. Traditional biogenic amine systems may respond to CER and diazepam administration in some compensatory manner.

Amantadine reduces haloperidol-induced dopamine receptor hypertensitivity in the striatum

In this report evidence is presented that amantadine hydrochloride greatly reduced the development of dopaminergic receptor hypersensitivity in the striatum, which normally results following chronic haloperidol administration using both a stereotyped behavior bioassay and a [3H]spiroperidol receptor binding assay. Amantadine prophylaxis reduced maximal ligand binding to near control levels and also significantly reduced apomorphine induced stereotypy. These results clearly demonstrate that amantadine greatly reduced haloperidol-induced striatal dopamine receptor hypersensitivity and support the hypothesis that amantadine given concurrently with neuroleptic agents might serve to prevent the development of human tardive dyskinesia.