Su-Jan Huang

Intracranial microdialysis of salicylic acid to detect hydroxyl radical generation through dopamine autooxidation in the caudate nucleus: effects of MPP+.

Ringers solution containing salicylic acid (5 nmol/microliters/min) was infused directly through an intracranial microdialysis probe to detect the generation of hydroxyl radicals (.OH) reflected by the formation of dihydroxybenzoic acids (DHBA) in the caudate nucleus of anesthetized rats. Brain dialysate was assayed for dopamine, 2,3-, and 2,5-DHBA by a high-pressure liquid chromatography-electrochemical (HPLC-EC) procedure. 1-Methyl-4-phenylpyridinium ions (MPP+, 0 to 150 nmol) increased dose-dependently the release of dopamine and the formation of DHBA. A positive linear correlation between the release of dopamine and the formation of 2,3- or 2,5-DHBA was observed (R2 = .98). The present results demonstrate the validity of the use of not only 2,3-DHBA but also 2,5-DHBA as an in vivo index of oxidative damage generated by reactive .OH radicals. In conclusion, the present study demonstrates a novel use of intracranial microdialysis of salicylic acid to assess the oxidative damage elicited by .OH in living brain.

Altered serotonin synthesis, turnover and dynamic regulation in multiple brain regions of mice lacking the serotonin transporter

To evaluate the consequences of inactivation of the serotonin transporter (SERT) gene on 5-HT homeostasis and function, 5-HT synthesis and turnover rates were measured using the decarboxylase inhibition method in multiple brain regions (frontal cortex, striatum, brainstem, hippocampus and hypothalamus) from mice with a genetic disruption of SERT. 5-HT synthesis rates were increased 30-60% in the different brain regions of SERT -/- mice compared to littermate +/+ control mice despite 55-70% reductions in tissue 5-HT concentrations. Brain regions that possessed a greater capacity to increase synthesis and turnover (frontal cortex, striatum) demonstrated lesser reductions in tissue 5-HT. Female SERT -/- mice had greater increases (79%) in brain 5-HT synthesis than male -/- mice did (25%), a finding associated with higher brain tryptophan concentrations in females. Despite increased 5-HT synthesis, there was no change in either TPH2 or TPH1 mRNA levels or in maximal in vitro TPH activity in the brainstem of SERT -/- mice. Catecholamine homeostasis as reflected in brain tissue concentrations and in synthesis and turnover of dopamine and norepinephrine was unchanged in SERT -/- mice. Taken together, the results demonstrate a markedly altered homeostatic situation in SERT -/- mice that lack 5-HT reuptake, resulting in markedly depleted tissue stores that are inadequately compensated for by increased 5-HT synthesis, with brain region and gender specificity observed.

Effects of isolation-rearing on locomotion, anxiety and responses to ethanol in Fawn Hooded and Wistar rats

Abstract Voluntary ethanol (EtOH) consumption is increased by isolation-rearing in several rat strains. The following experiments examined the effects of isolation-rearing on basal and ethanol-stimulated behavior in Fawn Hooded rats, an alcohol-preferring rat strain, compared to Wistar rats. Locomotor activity and anxiety were examined under both conditions. Basal locomotor activity was higher in isolated subjects of both strains in low light conditions, but under bright light conditions, this difference was only observed in Wistar rats. Locomotor stimulant effects of EtOH were only observed in isolation-reared rats. In the elevated plus maze, Fawn Hooded rats were more anxious than Wistar rats under low light conditions, but under bright light conditions, Wistar socials were less anxious than all of the other groups. Administration of 1.5 mg/kg EtOH produced an anxiolytic response in the elevated plus maze under bright light conditions in Fawn Hooded rats, but to a lesser degree Wistar rats, particularly Wistar isolates. In conclusion, although both strain and isolation-rearing had effects on locomotion and anxiety as well as the stimulatory and anxiolytic effects of EtOH, these effects appeared to be independent.

Effects of isolation-rearing on voluntary consumption of ethanol, sucrose and saccharin solutions in Fawn Hooded and Wistar rats

Abstract These experiments examined the hypothesis that isolation-rearing and strain influence hedonic mechanisms. In experiment 1, voluntary consumption of ethanol and water was monitored in the home cage of Fawn Hooded (FH) and Wistar rats. FH rats were found to consume more ethanol at low concentrations than Wistar rats, independent of rearing condition, and isolation-reared rats were found to consume more of high ethanol concentrations, independent of strain. In experiment 2, isolation-reared rats were found to consume more sucrose, independent of concentration, than socially reared rats. In experiment 3, Fawn Hooded rats were found to be more sensitive to low concentration solutions of saccharin, and to consume less of the high concentration solutions, while isolation-rearing was found to enhance consumption of high concentrations. Thus, hedonic processes are independently modulated by strain and rearing conditions, although the effects of isolation-rearing appear to be exacerbated in Fawn Hooded rats.

Differential basis of strain and rearing effects on open-field behavior in Fawn Hooded and Wistar rats.

Open-field behavior was examined under several conditions in isolation-reared, and socially reared, Fawn Hooded (FH) and Wistar rats. Lighting conditions (red or white light) and complexity (object or no object) were varied: Experiment 1, white light, no object; Experiment 2, red light, no object; Experiment 3, white light, object; Experiment 4, red light, object. The plasma corticosterone (CORT) response to open-field exposure was examined two further experiments. Observation of differences in open-field behavior, resulting from strain or rearing condition, was dependent on both lighting condition and complexity. Differences in exploratory behavior exhibited by isolation-reared rats were best explained by changes in response to novelty, while those in FH, relative to Wistar, rats were primarily due to increased anxiety. Supporting these conclusions, FH rats had enhanced stimulated CORT levels, while isolation rearing was without effect.

Effects of 5,7-Dihydroxytryptamine Depletion of Tissue Serotonin Levels on Extracellular Serotonin in the Striatum Assessed with In Vivo Microdialysis: Relationship to Behavior †

Effects of i.c.v. administration of 5,7‐dihydroxytryptamine (5,7‐DHT) on biochemistry and behavior were studied in awake Sprague‐Dawley rats. It was found that 5,7‐DHT depletion of striatal tissue levels of serotonin (5‐HT) does not diminish extracellular levels until substantial depletions occur. This finding is similar to those observed after 6‐hydroxydopamine lesions of the brain dopamine systems. Although varying amounts of 5,7‐DHT produced serotonin depletions in striatal tissue, decreases in extracellular levels were only observed at tissue depletions greater than 60% compared to saline‐injected control subjects. Thus, the effects of serotonin lesions which produce only moderate depletions may not be the result of decreased extracellular serotonin, but instead may be the result of compensatory changes in remaining neurons which maintain normal extracellular serotonin concentrations. Different degrees of striatal serotonin depletion were associated with opposite behavioral effects. Moderate levels of serotonin depletion (50–75%) produced evidence of increased anxiety, while these effects were no longer seen in rats with more severe 5‐HT depletions (>75%). Synapse 33:16–25, 1999.

Neurochemical, behavioral, and physiological effects of pharmacologically enhanced serotonin levels in serotonin transporter (SERT)-deficient mice

RationaleSerotonin transporter (SERT) knockout (−/−) mice have an altered phenotype in adulthood, including high baseline anxiety and depressive-like behaviors, associated with increased baseline extracellular serotonin levels throughout life.ObjectivesTo examine the effects of increases in serotonin following the administration of the serotonin precursor 5-hydroxy-l-tryptophan (5-HTP) in SERT wild-type (+/+), heterozygous (+/−), and −/− mice.Results5-HTP increased serotonin in all five brain areas examined with approximately 2- to 5-fold increases in SERT+/+ and +/− mice, and with greater 4.5- to 11.7-fold increases in SERT−/− mice. Behaviorally, 5-HTP induced exaggerated serotonin syndrome behaviors in SERT−/−, mice with similar effects in male and female mice. Studies suggest promiscuous serotonin uptake by the dopamine transporter (DAT) in SERT−/− mice, and here, the DAT blocker GBR 12909 enhanced 5-HTP-induced behaviors in SERT−/− mice. Physiologically, 5-HTP induced exaggerated temperature effects in SERT-deficient mice. The 5-HT1A antagonist WAY 100635 decreased 5-HTP-induced hypothermia in SERT+/+ and +/− mice with no effect in SERT−/− mice, whereas the 5-HT7 antagonist SB 269970 decreased this exaggerated response in SERT−/− mice only. WAY 100635 and SB 269970 together completely blocked 5-HTP-induced hypothermia in SERT+/− and −/− mice.ConclusionsThese studies demonstrate that SERT−/− mice have exaggerated neurochemical, behavioral, and physiological responses to further increases in serotonin, and provide the first evidence of intact 5-HT7 receptor function in SERT−/− mice, with interesting interactions between 5-HT1A and 5-HT7 receptors. As roles for 5-HT7 receptors in anxiety and depression were recently established, the current findings have implications for understanding the high anxiety and depressive-like phenotype of SERT-deficient mice.

The effects of social isolation on the forced swim test in Fawn hooded and Wistar rats.

Although the forced swim test (FST) has long been used as a preclinical screen of antidepressant efficacy, locomotor stimulants are known to produce confounding effects using the traditional dependent measure in this test: immobility. It has recently been suggested that measurement of struggling behavior may be a better index of antidepressant activity. The present experiments examined behavior in the forced swim test in two potential animal models of depression: the Fawn hooded rat, and the isolation-reared rat. No evidence was found to support these assertions, indeed immobility was decreased in Fawn hooded compared to Wistar rats, however this appeared to be caused by increased struggling behavior in Fawn hooded socials and increased swimming in Fawn hooded isolates. Although these differential results are highly suggestive of different underlying causes of decreased immobility in Fawn hooded rats depending on rearing conditions, the data suggests that the underlying psychological functions assumed to be represented by behavior assessed in this paradigm may not be adequately discriminated.

Delayed obsessive-compulsive disorder symptom exacerbation after a single dose of a serotonin antagonist in fluoxetine-treated but not untreated patients.

Abstract Enhanced serotonergic transmission may underlie therapeutic effects of serotonin reuptake inhibitors in obsessive-compulsive disorder. However, such treatment may decrease serotonin receptor responsivity. We investigated whether the serotonin antagonist metergoline would exacerbate or further improve systems in fluoxetine-responsive patients. Pilot results suggested open metergoline produced delayed symptom worsening in fluoxetine-treated patients. Fourteen patients continuing fluoxetine received metergoline and placebo (double-blind, randomized). Symptom ratings continued for 1 week afterwards. Ten unmedicated patients underwent the same procedures. Symptoms improved 4 h after both metergoline and placebo. The day after metergoline but not placebo, fluoxetine-treated patients had significantly increased anxiety, obsessions and compulsions, abating over several days. Depression was unchanged. Metergoline had no similar delayed effects in unmedicated patients. Metergoline levels were higher in fluoxetine-treated patients. These results, consistent with less conclusive earlier findings, suggest that prolonged changes in brain serotonin function underlie symptom re-emergence following administration of metergoline to fluoxetine-treated patients with obsessive-compulsive disorder.

Effects of Isolation‐Rearing on Acoustic Startle and Pre‐Pulse Inhibition in Wistar and Fawn Hooded Rats

Isolation-rearing of rats induces a variety of behavioral changes including impaired prepulse inhibition of acoustic startle’ and may potentiate acoustic startle (F.S.H., unpublished observations). Similar effects were reported for several psychiatric conditions, including posttraumatic stress disorder (PTSD).’ These behavioral changes in rats are associated with concomitant upregulation of presynaptic dopamine function and enhanced responses to dopamine agonists.’ One important question that has not yet been addressed is whether animals of different genetic backgrounds vary in sensitivity to the effects of early social deprivation. The present experiment examines the potential interaction between genetic and experiential effects on acoustic startle and pre-pulse inhibition (PPI) using Wistar and Fawn Hooded rats, to identify possible predisposing factors to augmentation of startle and impairment of PPI. Fawn Hooded rats have impaired serotonin function? as do isolation-reared rats.5 Under some conditions Fawn Hooded rats are also hyperactive (F.S.H., unpublished observation) which may indicate that they have enhanced dopamine function as well.