Extracellular 5-hydroxytryptamine Levels Research Articles

Luteolin suppresses 5-hydroxytryptamine elevation in stimulated RBL-2H3 cells and experimental colitis mice.

Increased 5-hydroxytryptamine may be associated with the development and progression of inflammatory bowel disease. In this study, we examined the suppressive effect of flavonoids on the increased intra- and extracellular 5-hydroxytryptamine levels in rat mast RBL-2H3 cells, known to produce 5-hydroxytryptamine by the phorbol 12-myristate 13-acetate stimulation. Among the flavonoids examined, luteolin and quercetin significantly reduced the cellular 5-hydroxytryptamine concentration. Gene and protein expression analyses revealed that luteolin significantly suppressed cellular tryptophan hydroxylase 1 expression induced by phorbol 12-myristate 13-acetate stimulation. Mitogen-activated protein kinase/extracellular signal-regulated kinase signaling was also suppressed by luteolin, suggesting that this pathway is one of targets of 5-hydroxytryptamine modulation by luteolin. An in vivo experimental colitis model was prepared by administering 2.5% dextran sodium sulfate in drinking water to C57BL/6 mice for seven days. The ingestion of 0.1% dietary luteolin suppressed the increasing 5-hydroxytryptamine in the colorectal mucosa. In conclusion, luteolin possesses a suppressive effect on extensive 5-hydroxytryptamine formation in both experimental RBL-2H3 cells and colitis models.

Acute morphine administration increases extracellular DA levels in the rat lateral septum by decreasing the GABAergic inhibitory tone in the ventral tegmental area

We studied the effect of an acute systemic administration of morphine and of a local intra-ventral tegmental area (VTA) infusion of the same drug on extracellular levels of dopamine (DA) in the lateral septum (LS) by in vivo microdialysis in anesthetized rats. The extracellular levels of 5-hydroxytryptamine (5-HT) were also measured in all dialysate samples. The acute systemic administration of morphine dose-dependently increased extracellular levels of DA but not of 5-HT in the LS, in the absence or presence of fluoxetine. This morphine effect was antagonized by the previous administration of naloxone, a specific opioid antagonist. The local infusion of morphine in the VTA also induced a significant increase of the extracellular levels of DA in the LS, concomitantly with a decrease of gamma-aminobutyric acid (GABA) extracellular levels in the VTA itself. Intriguingly, the LS extracellular levels of DA returned to basal values before the VTA GABA extracellular levels recovered. Our results show for the first time that an acute administration of morphine increases DA extracellular levels in the LS. The results also suggest that DA cells in the VTA and innervating the LS are under an inhibitory GABAergic tone sensitive to morphine. Taken together, our neurochemical data and previous studies involving LS DA in stress-related behavior support the hypothesis that DA in the LS plays a significant role in addictive behavior. The participation of LS DA and 5-HT systems in stress-induced relapse to drug seeking should be studied further.

LY393558, a 5-hydroxytryptamine reuptake inhibitor and 5-HT 1B/1D receptor antagonist: effects on extracellular levels of 5-hydroxytryptamine in the guinea pig and rat

The stimulation of terminal 5-HT 1B/1D autoreceptors limits the effects of selective serotonin reuptake inhibitors on extracellular levels of 5-hydroxytryptamine (5-HT, serotonin) in vivo. Microdialysis studies show that acute oral administration of LY393558—a 5-HT reuptake inhibitor and antagonist at both the human 5-HT 1B and 5-HT 1D receptor—in the dose range 1–20 mg/kg, increases extracellular levels of 5-HT in both the guinea pig hypothalamus and rat frontal cortex. In both species, the levels of 5-HT that were attained were higher than following an acute, maximally effective dose of fluoxetine (20 mg/kg orally), reaching approximately 1500% in the guinea pig hypothalamus and 700% in the rat frontal cortex. In both species, the response to LY393558 (10 mg/kg p.o.) was impulse dependent, being absent in the presence of tetrodotoxin delivered at 1 μM via the microdialysis probe. The sensitivity to tetrodotoxin contrasted with the effects seen with dl-fenfluramine. Studies in rats showed that the microdialysate 5-HT concentration achieved in the frontal cortex after an acute challenge with LY393558 (5 mg/kg p.o.) was significantly greater than following a chronic regime of fluoxetine treatment (10 mg/kg/day orally for 21 days). Moreover, in rats chronically treated with LY393558 (5 mg/kg/day orally for 21 days), the mean basal concentration, 24 h after the final pretreatment dose, was of the same magnitude as that following chronic fluoxetine. However, in contrast to the response seen in fluoxetine-pretreated animals, a challenge dose of LY393558 still elicited a further increase in extracellular 5-HT in LY393558-pretreated animals. LY393558 is a potent 5-HT reuptake inhibitor and 5-HT 1B/1D receptor antagonist. Microdialysis studies show that acute oral administration increases extracellular levels of 5-HT, by an impulse-dependent mechanism, above those produced by a maximally effective dose of fluoxetine, and in rats to levels only achieved following chronic fluoxetine treatment. Its neurochemical profile in vivo suggests that it may be a more effective antidepressant with the potential for producing an earlier onset of clinical activity than selective serotonin reuptake inhibitors.

Desensitisation of 5-HT autoreceptors upon pharmacokinetically monitored chronic treatment with citalopram

Rats were chronically treated with the selective serotonin re-uptake inhibitor citalopram [1-(3-dimethylaminopropyl)-1-(4-fluorophenyl)-5-phtalancarbonitril], by means of osmotic minipumps. Using an infusion concentration of 50 mg/ml citalopram, steady-state plasma concentrations of approximately 0.3 μM citalopram were maintained for 15 days. Citalopram plasma levels dropped below pharmacologically active concentrations 48 h after removal of the minipumps. Although chronic treatment with citalopram did induce an attenuated response by extracellular levels of 5-hydroxytryptamine (5-HT) after systemic administration of the 5-HT 1A receptor agonist 8-hydroxy-2-(di- n-propylamino)tetralin (8-OH-DPAT), no effect of chronic citalopram treatment was observed when 5-HT 1B receptor function was evaluated with a local infusion of 5-HT 1B/D receptor agonist, sumatriptan (3-[2-dimethylamino]ethyl- N-methyl-1 H-indole-5methane sulphonamide). Controversially, no augmentation of the increase of 5-HT levels was observed upon systemic administration of citalopram. It is concluded that, although chronic treatment with citalopram does induce desensitisation of 5-HT 1A receptors, the absence of augmented effects of citalopram on 5-HT levels indicates that other mechanisms compensate for the loss of autoreceptor control.

Formula omitted] exchanger in rat hippocampal neurons produces a slow negative DC potential after in vitro ischemia

We have investigated the effects of infusing the excitatory amino acid agonist α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) on extracellular levels of 5-hydroxytryptamine (5-HT) and 5-hydoxyindoleacetic acid (5-HIAA) in rat striatum using in vivo microdialysis. AMPA (50–500 μM) caused a concentration-dependent increase in extracellular 5-HT, while having the converse effect on 5-HIAA. At the highest agonist dose the decrease in dialysate 5-HIAA was followed by a significant increase in this metabolite. Two hundred micromolar 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), a competitive non-NMDA glutamate receptor antagonist, reversed the effects of a 100 μM AMPA on dialysate 5-HT and 5-HIAA. Co-infusion of AMPA with tetrodotoxin (TTX) abolished the effects of 100 μM AMPA, but only partially reversed the effect of 500 μM AMPA on 5-HT release. We have also investigated whether AMPA receptor desensitization, a well documented event, plays a role in AMPA receptor modulation of striatal 5-HT release. Diazoxide (500 μM), a drug which prevents AMPA receptor desensitization, failed to augment the effect of 100 μM AMPA on 5-HT release. Diazoxide alone significantly decreased 5-HT release, as did the drug cromakalim (100 μM), probably as a result of their common action as activators of ATP-dependent K+ channels. It is concluded that AMPA receptors play a role in regulating both 5-HT release and metabolism in rat striatum. However, AMPA receptor desensitization does not appear to play a role in this process in this structure.

The Effect of Repeated Exposure to Forced Swimming on Extracellular Levels of 5-Hydroxytryptamine in the Rat

The effects of repeated exposure to forced swimming was examined on extracellular concentrations of 5-hydroxytryptamine (5-HT), behavioral, and endocrine responses in rats. Animals were exposed to a 15-min swimming session on two consecutive days. On the first day, the swimming session increased extracellular 5-HT by 80% over baseline in the striatum and reduced 5-HT to 40% below baseline in the lateral septum. On the second day, however, the swimming session produced no effect on 5-HT in either brain region. Ratings of behavior showed that rats demonstrated climbing and swimming behaviors and developed immobility during the initial swimming session and that ratings of immobility increased and swimming decreased during the second swimming session. Immobility was positively correlated and swimming was negatively correlated with changes in extracellular 5-HT in the lateral septum but not in the striatum. Plasma corticosterone was equally elevated by 950 % after either 1 or 2 days of swimming exposure. These results show that there is rapid adaptation to the effects of repeated forced swimming on the regionally-specific, bi-directional response of extracellular 5-HT. In addition, changes in extracellular 5-HT in the lateral septum may be related to the behaviors produced during the forced swimming test that underlie its utility as an animal model of depression.

On-line detection of extracellular levels of serotonin in dorsal raphe nucleus and frontal cortex over the sleep/wake cycle in the freely moving rat

We used in vivo microdialysis coupled with polygraphic recording to monitor 5-hydroxytryptamine levels in the dorsal raphe nucleus and frontal cortex across waking, slow-wave sleep and rapid eye-movement sleep. Male Sprague–Dawley rats were prepared with electroencephalogram and electromyogram electrodes. Microdialysis probes were placed in dorsal raphe nucleus and/or frontal cortex. Dialysate samples were manually collected during polygraphically-defined behavioural states and the level of serotonin was assayed by means of microbore high-performance liquid chromatography separation and electrochemical detection. Samples from microdialysis probes histologically localized to the dorsal raphe nucleus and frontal cortex showed different levels of extracellular 5-hydroxytryptamine in waking, slow-wave sleep and rapid eye-movement sleep. In dorsal raphe nucleus the extracellular level of serotonin was highest in waking, decreased in slow-wave sleep to 69% and in rapid eye-movement sleep to 39% of waking mean level (waking 3.2±0.9; slow-wave sleep 2.2±0.8; rapid eye-movement sleep 1.3±0.4 fmol/sample). Mean extracellular levels of serotonin in frontal cortex displayed a similar pattern (waking 1.7±0.4; slow-wave sleep 1.0±0.3; rapid eye-movement 0.5±0.05 fmol/sample). In frontal cortex, rapid eye-movement sleep samples were only obtained in three animals. Our findings are consistent with previous results in cats, and suggest that in rats also, extracellular 5-hydroxytryptamine levels in dorsal raphe nucleus and frontal cortex across the sleep/wake cycle might reflect serotonergic neuronal activity. The findings stress the importance of controlling for behavioural state when investigating neurochemical correlates of serotonergic function.

Conditioned release of 5-hydroxytryptamine in vivo in the nucleus accumbens following isolation-rearing in the rat

This study examined the effect of isolation-rearing in the Lister hooded rat on extracellular 5-hydroxytryptamine in the medial nucleus accumbens following footshock and in relation to a conditioned emotional response. Inescapable mild footshock was associated with an immediate and prolonged increase in extracellular 5-hydroxytryptamine in the medial nucleus accumbens of isolation-reared rats. In group-reared rats (footshock-treated) and control groups (no footshock) there was no significant change in extracellular 5-hydroxytryptamine levels. When exposed to the contextual stimulus 140min later (testing box without shock) there was an immediate and long-lasting increase in extracellular 5-hydroxytryptamine in the nucleus accumbens of the isolation-reared rats, however, the contextual stimulus did not significantly affect extracellular 5-hydroxytryptamine in the medial nucleus accumbens of group-reared rats.The results show that exposure to footshock and conditioning to context are not normally associated with a change in extracellular 5-hydroxytryptamine in the medial nucleus accumbens, however, in rats exposed to social isolation from weaning, both stimuli increase extracellular 5-hydroxytryptamine. The isolation-induced increase in presynaptic serotonergic function in the medial nucleus accumbens contrasts with previous reports of reduced 5-hydroxytryptamine release in the hippocampus and therefore suggests that isolation-rearing differentially affects the function of serotonergic neurons in the brain. The changes in 5-hydroxytryptamine function in the medial nucleus accumbens may represent physiological adaptations to stress or may occur secondary to changes in the function of another neurotransmitter, possibly dopamine.

AMPA receptors modulate extracellular 5-hydroxytryptamine concentration and metabolism in rat striatum in vivo

We have investigated the effects of infusing the excitatory amino acid agonist α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) on extracellular levels of 5-hydroxytryptamine (5-HT) and 5-hydoxyindoleacetic acid (5-HIAA) in rat striatum using in vivo microdialysis. AMPA (50–500 μM) caused a concentration-dependent increase in extracellular 5-HT, while having the converse effect on 5-HIAA. At the highest agonist dose the decrease in dialysate 5-HIAA was followed by a significant increase in this metabolite. Two hundred micromolar 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), a competitive non-NMDA glutamate receptor antagonist, reversed the effects of a 100 μM AMPA on dialysate 5-HT and 5-HIAA. Co-infusion of AMPA with tetrodotoxin (TTX) abolished the effects of 100 μM AMPA, but only partially reversed the effect of 500 μM AMPA on 5-HT release. We have also investigated whether AMPA receptor desensitization, a well documented event, plays a role in AMPA receptor modulation of striatal 5-HT release. Diazoxide (500 μM), a drug which prevents AMPA receptor desensitization, failed to augment the effect of 100 μM AMPA on 5-HT release. Diazoxide alone significantly decreased 5-HT release, as did the drug cromakalim (100 μM), probably as a result of their common action as activators of ATP-dependent K + channels. It is concluded that AMPA receptors play a role in regulating both 5-HT release and metabolism in rat striatum. However, AMPA receptor desensitization does not appear to play a role in this process in this structure.

Regional differences in the effects of forced swimming on extracellular levels of 5-hydroxytryptamine and 5-hydroxyindoleacetic acid

The effects of forced swimming for 30 min on extracellular 5-hydroxytryptamine (5-HT) levels were examined in five brain regions in rats using in vivo microdialysis. A single dialysis probe was implanted under surgical anesthesia into either the striatum, ventral hippocampus, frontal cortex, amygdala, or lateral septum on the day before the study. Dialysate content of 5-HT and 5-hydroxyindoleacetic acid (5-HIAA) was measured by HPLC. Forced swimming elevated extracellular levels of 5-HT in the striatum to a maximum of 90% above baseline. In contrast, forced swimming reduced 5-HT levels in the amygdala and lateral septum to 50 and 40% of baseline, respectively. In the hippocampus and frontal cortex, 5-HT levels were not altered significantly by forced swimming. In all five brain regions, forced swimming reduced 5-HIAA levels to 45–60% of baseline. These results suggest that forced swimming modulates 5-HT neurotransmission in a regionally specific manner. Aside from being a significant biological stressor, the forced swimming test is used as an animal behavioral model to detect antidepressant drugs, including drugs that alter 5-HT neurotransmission. It is possible that the alterations of extracellular levels of 5-HT produced by forced swimming in certain brain regions may be associated with the ability of antidepressant drugs to selectively alter behavioral performance during the forced swimming test.

Substance P injection into the dorsal raphe increases blood pressure and serotonin release in hippocampus of conscious rats

Microinjections of substance P (SP, 100 pmol) into the dorsal raphe nucleus (DRN) in conscious rats increased blood pressure and heart rate for 30–40 min. Concomitantly, the extracellular levels of 5-hydroxytryptamine (5-HT) in the ventral hippocampus, monitored by microdialysis, increased by 30% for 20 min compared with the vehicle control. Pretreatment with the 5-HT 2 receptor antagonist, ritanserin (1 mg/kg i.v.), prevented the pressor response to SP but not the increase in heart rate. Pretreatment with the partial 5-HT 1A receptor agonist, 8-methoxy-2-(N-2-chloroethyl-N-n-propyl)amino tetralin (8-MeO-CLEPAT, 10 μg/kg i.v.) prevented the increase in both blood pressure and heart rate. It is suggested that microinjections of SP into the DRN increase blood pressure through activation of serotonergic DRN neurons and that the postsynaptic receptor responsible for the pressor response is of the 5-HT 2 type.

Measurement of extracellular 5-HT and 5-HIAA in hippocampus of freely moving rats using microdialysis: long-term applications

Extracellular levels of 5-hydroxytryptamine (5-HT) and its metabolite 5-hydroxyindoleacetic acid (5-HIAA) were measured in the ventral hippocampus of the awake rat using microdialysis. The basal level of 5-HT in hippocampal dialysates was very close to the detection limit of our assay. However, addition of a 5-HT re-uptake blocker, citalopram, to the perfusion medium resulted in a 3-fold rise of 5-HT levels in dialysates and provided a stable baseline. Under these conditions, extracellular levels of 5-HT and 5-HIAA could be measured up to 11 days after dialysis probe implantation. 5-HT measured in dialysate was Ca(2+)-dependent both 24 h and 7 days after surgery. In comparison K(+)-induced depolarization caused a 9-fold increase in 5-HT output 24 h after probe implantation and this effect had disappeared 10 days later, although behavioral activation following K(+) application was similar 24 h and 11 days after surgery. Systematic administration of l-trytophan had no significant effect on 5-HT but increased 5-HIAA levels by 90%. These results suggest that in the presence of citalopram, 5-HT in hippocampal dialysates is derived from serotonergic neurones. The microdialysis method appears most useful for measurement of extracellular 5-HT 24-72 h after probe implantation. Finally, our data indicate that extracellular 5-HIAA mainly reflects intraneuronal metabolism of 5-HT prior to release.