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2.
J Neurochem ; 101(3): 641-51, 2007 May.
Article in English | MEDLINE | ID: mdl-17254018

ABSTRACT

In the present study, we used high-speed chronoamperometry to examine serotonin (5-HT) transporter (5-HTT) function in vivo in 2-, 5-, and 10-month-old brain-derived neurotrophic factor (BDNF)+/- mice. The rate of clearance of exogenously applied 5-HT was measured in CA3 region of hippocampus. In 2-month-old mice, the rate of 5-HT clearance did not differ between BDNF+/+ and BDNF+/- mice. In BDNF+/+ mice, 5-HT clearance rate (Tc) increased markedly with age. In contrast, Tc remained relatively static in BDNF+/- mice across 2-, 5-, and 10-month age groups. At 5 months of age, female BDNF+/+ mice had a lower maximal velocity (Vmax) for 5-HT clearance than male BDNF+/+ mice. There was a similar trend in 5-month-old BDNF+/- mice, but this did not reach statistical significance. There was an age-dependent increase in KT value for 5-HT clearance (i.e., decreased in vivo affinity of 5-HTT), but no significant effect of genotype or gender. 5-HTT density, as measured by [3H]cyanoimipramine binding, was not different between BDNF+/+ and BDNF+/- mice, although there was a significant increase in 5-HTT binding with age. The selective 5-HT reuptake inhibitor fluvoxamine (50 and 100 pmol) significantly decreased 5-HT clearance in BDNF+/+ mice, but not in BDNF+/- mice. Our data suggest that the profoundly reduced ability of 5- and 10-month-old BDNF+/- mice to clear 5-HT is not because of a decrease in the total number of 5-HTTs, but may be due to functional deficits in the 5-HTT, e.g., in the machinery/signaling required for insertion of 5-HTTs into the plasma membrane and/or activation of the 5-HTT once it is positioned to take up 5-HT from extracellular fluid.


Subject(s)
Brain-Derived Neurotrophic Factor/physiology , Gene Expression/physiology , Serotonin Plasma Membrane Transport Proteins/physiology , Age Factors , Animals , Autoradiography/methods , Brain-Derived Neurotrophic Factor/deficiency , Dose-Response Relationship, Drug , Electrochemistry/methods , Female , Fluvoxamine/pharmacology , Gene Expression/drug effects , Hippocampus/drug effects , Hippocampus/metabolism , Imipramine/analogs & derivatives , Imipramine/pharmacokinetics , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Protein Binding/drug effects , Serotonin/metabolism , Serotonin/pharmacology , Selective Serotonin Reuptake Inhibitors/pharmacology , Sex Factors
3.
Toxicol Pathol ; 33(6): 702-10, 2005.
Article in English | MEDLINE | ID: mdl-16243775

ABSTRACT

Factor XIII (FXIII) is a thrombin-activated plasma coagulation factor critical for blood clot stabilization and longevity. Administration of exogenous FXIII to replenish depleted stores after major surgery, including cardiopulmonary bypass, may reduce bleeding complications and transfusion requirements. Thus, a model of extracorporeal circulation (ECC) was developed in adult male cynomolgus monkeys (Macaca fascicularis) to evaluate the nonclinical safety of recombinant human FXIII (rFXIII). The hematological and coagulation profile in study animals during and after 2 h of ECC was similar to that reported for humans during and after cardiopulmonary bypass, including observations of anemia, thrombocytopenia, and activation of coagulation and platelets. Intravenous slow bolus injection of 300 U/kg (2.1 mg/kg) or 1000 U/kg (7 mg/kg) rFXIII after 2 h of ECC was well tolerated in study animals, and was associated with a dose-dependent increase in FXIII activity. No clinically significant effects in respiration, ECG, heart rate, blood pressure, body temperature, clinical chemistry, hematology (including platelet counts), or indicators of thrombosis (thrombin:anti-thrombin complex and D-Dimer) or platelet activation (platelet factor 4 and beta-thromboglobulin) were related to rFXIII administration. Specific examination of brain, heart, lung, liver, and kidney from rFXIII-treated animals provided no evidence of histopathological alterations suggestive of subclinical hemorrhage or thrombosis. Taken as a whole, the results demonstrate the ECC model suitably replicated the clinical presentation reported for humans during and after cardiopulmonary bypass surgery, and do not suggest significant concerns regarding use of rFXIII in replacement therapy after extracorporeal circulation.


Subject(s)
Blood Coagulation Disorders/drug therapy , Cardiopulmonary Bypass , Coagulants/therapeutic use , Extracorporeal Circulation , Factor XIII/therapeutic use , Postoperative Complications/drug therapy , Animals , Blood Coagulation/drug effects , Blood Coagulation Disorders/blood , Coagulants/administration & dosage , Disease Models, Animal , Dose-Response Relationship, Drug , Drug Evaluation, Preclinical , Factor XIII/administration & dosage , Humans , Macaca fascicularis , Male , Pilot Projects , Platelet Activation/drug effects , Postoperative Care , Postoperative Complications/blood , Recombinant Proteins/therapeutic use , Time Factors
4.
J Neurochem ; 90(5): 1269-75, 2004 Sep.
Article in English | MEDLINE | ID: mdl-15312182

ABSTRACT

Valproic acid (VPA), which has demonstrated efficacy in the treatment of bipolar disorder, has been shown to alter components of the phosphoinositide (PI) signaling cascade and to increase gene expression mediated by the transcription factor activator protein 1 (AP-1). Central serotonin-2A (5-HT2A) receptors, which have been implicated in the pathophysiology of manic-depressive illness, are coupled to PI hydrolysis. The promoter region of the 5-HT2A receptor gene contains AP-1 binding sites. We examined in C6 glioma cells the effect of VPA on 5-HT2A receptor signaling. Treatment of cells with VPA (100 microg/mL) for 20 h, but not 1.5 h, resulted in an enhancement of 5-HT2A receptor-stimulated PI hydrolysis. This effect of 20-h VPA exposure appeared not to be at the level of G protein or effector (i.e. phospholipase C: PLC) as inositol phosphate accumulation stimulated by aluminum fluoride or the PLC activator 2,4,6-trimethyl-N-(m-3-trifluromethylphenyl) benzenesulfonamide was not increased. The number of 5-HT2A receptors, as determined in saturation binding experiments using [3H]ketanserin, was increased by 20-h VPA treatment, with no change in affinity (KD). Taken together, our data suggest that the increase in 5-HT2A receptor-mediated PI hydrolysis following 20-h VPA exposure is not due to a general effect of VPA on this signaling cascade, but due to the up-regulation of 5-HT2A receptor number.


Subject(s)
Receptor, Serotonin, 5-HT2A/physiology , Signal Transduction/drug effects , Valproic Acid/pharmacology , Animals , Anticonvulsants/pharmacology , Binding Sites/drug effects , Cell Line , Dose-Response Relationship, Drug , Glioma/pathology , Ketanserin/pharmacokinetics , Mice , Phosphatidylinositols/metabolism , Quipazine/pharmacology , Signal Transduction/physiology , Time Factors , Tritium/pharmacokinetics
5.
J Neurochem ; 85(5): 1139-47, 2003 Jun.
Article in English | MEDLINE | ID: mdl-12753073

ABSTRACT

Heterozygous brain-derived neurotrophic factor (BDNF) (+/-) mice display abnormalities in central serotonergic neurotransmission, develop decrements in serotonergic innervation of the forebrain, and exhibit enhanced intermale aggressiveness. As disturbances of serotonin neurotransmission are implicated in alcohol abuse and aggression, we have examined in BDNF (+/-) mice alcohol drinking behavior, as well as central 5-hydroxytryptamine (5-HT)1A receptor function at the level of 5-HT1A receptor-G protein interaction. BDNF (+/-) mice displayed increased ethanol intake in a two-bottle choice procedure. There was no difference in the preference ratio for non-alcoholic tastants (i.e. quinine or saccharin) between genotypes. In the brains of alcohol-naive mice, we measured [35S]GTP gamma S binding stimulated by the 5-HT1A receptor agonist (+/-)-8-hydroxy-2-dipropyl-aminotetralin hydrobromide (8-OH-DPAT; 1 microM). In BDNF (+/-) versus wild-type (WT) mice, 5-HT1A receptor-stimulated [35S]GTP gamma S binding was significantly attenuated in the median raphe nucleus. There was a decrease in (+/-)8-OH-DPAT-stimulated [35S]GTP gamma S binding in the dorsal raphe, which did not reach statistical significance. In the hippocampus, 5-HT1A receptor-stimulated [35S]GTP gamma S binding was significantly attenuated in BDNF (+/-) mice. 5-HT1A receptor-stimulated [35S]GTP gamma S binding was attenuated in the anterior cingulate cortex and lateral septum, although these reductions did not reach statistical significance. 5-HT1A receptor number was not different between genotypes in any area of brain examined, suggesting that 5-HT1A receptor function, specifically the capacity of the 5-HT1A receptor to activate G proteins, is attenuated in BDNF (+/-) mice.


Subject(s)
Alcohol Drinking , Brain-Derived Neurotrophic Factor/deficiency , Ethanol/administration & dosage , Receptors, Serotonin/metabolism , 8-Hydroxy-2-(di-n-propylamino)tetralin/pharmacology , Alcohol Drinking/genetics , Animals , Autoradiography , Behavior, Animal/drug effects , Binding, Competitive/drug effects , Brain-Derived Neurotrophic Factor/genetics , Choice Behavior/drug effects , Female , GTP-Binding Proteins/metabolism , Guanosine 5'-O-(3-Thiotriphosphate)/pharmacokinetics , Heterozygote , Hippocampus/drug effects , Hippocampus/metabolism , Mice , Mice, Inbred C57BL , Mice, Mutant Strains , Raphe Nuclei/drug effects , Raphe Nuclei/metabolism , Receptors, Serotonin/drug effects , Receptors, Serotonin, 5-HT1 , Self Administration , Serotonin Receptor Agonists/pharmacology
6.
Brain Res ; 957(1): 174-82, 2002 Dec 06.
Article in English | MEDLINE | ID: mdl-12443993

ABSTRACT

Previous studies have shown that administration of the 5-HT(2) receptor agonist DOI to rats results in the heterologous desensitization of 5-HT(1A) receptor-mediated behavioral and neuroendocrine responses [Neuropsychopharmacology 19 (1998) 354; J. Neurosci. 21 (2001) 7919]. We hypothesized that the basis for these changes in 5-HT(1A) receptor function may involve changes in the capacity of the 5-HT(1A) receptor to activate G proteins. We examined the effect of chronic administration of DOI on the regulation of 5-HT(1A) receptor function at the level of receptor-G protein interaction using quantitative autoradiography of [(35)S]GTPgammaS binding stimulated by the 5-HT(1A) receptor agonist (+/-)8-OH-DPAT (1 microM). Repeated administration of DOI (1 mg/kg, s.c. once daily for 8 days) resulted in a marked down-regulation in 5-HT(2A) binding sites, as labeled by the antagonist radioligand [(3)H]ketanserin, throughout the cerebral cortex. Chronic DOI treatment also resulted in a significant and selective attenuation of 5-HT(1A) receptor-stimulated [(35)S]GTPgammaS binding in the anterior cingulate cortex (vehicle-treated: 74+/-7.7% above basal; DOI-treated: 43+/-4.6% above basal). Interestingly, 5-HT(1A) receptor-stimulated [(35)S]GTPgammaS binding was not altered in the dorsal or median raphe, or in the limbic structures and other cortical regions examined. The decrease in 5-HT(1A) receptor-stimulated [(35)S]GTPgammaS binding in anterior cingulate cortex was not due to a decrease in 5-HT(1A) receptor number, indicating that the capacity of the 5-HT(1A) receptor to activate G proteins is attenuated in this cortical area following repeated DOI treatment. The heterologous regulation of 5-HT(1A) receptor function by chronic 5-HT(2) receptor activation in the anterior cingulate cortex raises interesting questions as to how the regulatory interaction between these serotonin receptor subtypes influences cognition, memory and emotion.


Subject(s)
Amphetamines/pharmacology , Brain/metabolism , Guanosine 5'-O-(3-Thiotriphosphate)/metabolism , Receptors, Serotonin/metabolism , Serotonin Receptor Agonists/pharmacology , Animals , Autoradiography , Brain/drug effects , Gyrus Cinguli/metabolism , Ketanserin/pharmacology , Male , Rats , Rats, Sprague-Dawley , Receptor, Serotonin, 5-HT2A , Receptors, Serotonin, 5-HT1 , Serotonin Antagonists/pharmacology , Sulfur Radioisotopes
7.
J Neurochem ; 77(2): 589-97, 2001 Apr.
Article in English | MEDLINE | ID: mdl-11299321

ABSTRACT

We have investigated in C6 glioma cells the involvement of protein kinase C (PKC) in the regulation of serotonin-(2A) receptor (5-HT(2A) receptor) expression by agonist treatment. Comparison of the time-courses of agonist-induced downregulation of receptor number and mRNA indicate that a decrease in the number of 5-HT(2A) receptor binding sites in response to serotonin (5-HT) treatment is preceded by a decrease in 5-HT(2A) receptor mRNA. This decrease in 5-HT(2A) receptor mRNA as a result of agonist exposure was not due to a change in the stability or half-life of the transcript. Pretreatment of cells with the PKC inhibitor bisindolylmaleimide blocked the decrease in 5-HT(2A) receptor mRNA levels, and attenuated the down-regulation of 5-HT(2A) receptor binding sites induced by treatment with 5-HT. Experiments performed with the PKC inhibitors calphostin C and Gö 6976 confirmed that PKC was involved in the regulation of 5-HT(2A) receptor mRNA by agonist and implicate the conventional subgroup of PKC isoforms. Western blot analysis, using isoform-specific anti-PKC antibodies showed that under our culture conditions C6 glioma cells express the conventional isoforms PKC alpha, PKC gamma, as well as the novel isoforms PKC delta, PKC epsilon, and the atypical isoforms PKC lambda and PKC iota. Upon treatment with 5-HT for 10 min levels of the conventional isoforms PKC alpha and PKC gamma increased in the nuclear fraction. Taken together, our results implicate PKC alpha and/or PKC gamma in the regulation of 5-HT(2A) mRNA receptor and binding sites in response to agonist treatment.


Subject(s)
Down-Regulation/drug effects , Gene Expression Regulation/physiology , Protein Kinase C/physiology , Receptors, Serotonin/biosynthesis , Animals , Binding Sites , Carbazoles/pharmacology , Enzyme Induction/drug effects , Gene Expression Regulation/drug effects , Gene Expression Regulation, Neoplastic/drug effects , Glioma/pathology , Indoles/pharmacology , Maleimides/pharmacology , Mice , Naphthalenes/pharmacology , Neoplasm Proteins/antagonists & inhibitors , Neoplasm Proteins/biosynthesis , Neoplasm Proteins/genetics , Neoplasm Proteins/physiology , Protein Isoforms/antagonists & inhibitors , Protein Isoforms/physiology , Protein Kinase C/antagonists & inhibitors , RNA, Messenger/biosynthesis , RNA, Neoplasm/biosynthesis , Receptor, Serotonin, 5-HT2A , Receptors, Serotonin/genetics , Serotonin/pharmacology , Signal Transduction/drug effects , Tumor Cells, Cultured/enzymology
8.
Hypertension ; 37(2 Pt 2): 614-8, 2001 Feb.
Article in English | MEDLINE | ID: mdl-11230344

ABSTRACT

The goal of this study was to determine whether gamma-aminobutyric acid (GABA)ergic transmission and GABA binding are altered in chronic renal-wrap hypertension. Three groups of hypertensive and sham-operated rats were prepared for separate protocols. Four weeks later, the animals were prepared with femoral artery catheters for the measurement of mean arterial pressure. In all groups, blood pressure was significantly higher in the renal-wrapped animals. In the first study, bilateral microinjection of the GABA-A antagonist, bicuculline (50 pmol/site), into the paraventricular nucleus of the hypothalamus (PVN) caused a greater increase in arterial pressure (21.9+/-1.4 versus 16.7+/-1.8 mm Hg, P<0.05) and heart rate (135+/-15 versus 98+/-12 bpm, P=0.064) in hypertensive rats. [(3)H]Flunitrazepam was used to measure binding to the GABA-A receptor. Magnocellular neurons and the adjacent medial parvicellular neurons had more intense binding compared with the remainder of the PVN. B(max) was greater for the higher density binding area; the K(d) value was less in the high-density region. There were no differences in these parameters between normotensive and hypertensive animals. Competitive reverse transcription-polymerase chain reaction was used to measure the expression of mRNA for the alpha(1) subunit of the GABA-A receptor. No difference was observed in the mRNA between renal-wrapped and sham-operated rats. In summary, inhibition of GABA-A receptors in the PVN is augmented in the chronic phase of hypertension and is unrelated to a change in the expression of the number or affinity to the receptor. These findings suggest that the greater GABAergic activity is the result of an increase in GABA release in the PVN in chronic renal-wrap hypertension.


Subject(s)
Hypertension, Renovascular/metabolism , Paraventricular Hypothalamic Nucleus/metabolism , gamma-Aminobutyric Acid/metabolism , Animals , Bicuculline/pharmacology , Binding Sites , Blood Pressure/drug effects , Chronic Disease , GABA Antagonists/pharmacology , GABA-A Receptor Antagonists , Heart Rate/drug effects , Hypertension, Renovascular/blood , Male , Microinjections , Norepinephrine/blood , RNA, Messenger/metabolism , Radioligand Assay , Rats , Rats, Sprague-Dawley , Receptors, GABA-A/metabolism , Reverse Transcriptase Polymerase Chain Reaction
9.
Br J Pharmacol ; 132(2): 605-11, 2001 Jan.
Article in English | MEDLINE | ID: mdl-11159711

ABSTRACT

1. Because changes 5-HT(1A) receptor number do not occur following repeated agonist treatment, we hypothesized that the basis for 5-HT(1A) receptor desensitization involves changes in receptor-G protein coupling. We measured the effect of repeated agonist administration on 5-HT(1A) receptor-stimulated [(35)S]-GTPgammaS binding in forebrain areas, (i.e. anterior cingulate cortex, lateral septum, hippocampus, entorhinal cortex), and serotonergic cell body areas, the dorsal and median raphe nuclei. 2. Following treatment of rats with (+/-)8-OH-DPAT (1 mg kg(-1), s.c.) for 7 or 14 days, 5-HT(1A) receptor-stimulated [(35)S]-GTPgammaS binding was significantly attenuated in both the dorsal and median raphe nuclei. 3. 5-HT(1A) receptor-stimulated [(35)S]-GTPgammaS binding was significantly attenuated in the CA(1) region of the hippocampus after 7, but not 14 days of 8-OH-DPAT administration. 5-HT(1A) receptor-stimulated [(35)S]-GTPgammaS binding was not altered in other forebrain areas examined. 4. The binding of [(3)H]-MPPF to 5-HT(1A) receptor sites was not altered in any brain region examined following repeated agonist administration, suggesting that the observed changes in (+/-)8-OH-DPAT-stimulated [(35)S]-GTPgammaS binding were not due to changes in 5-HT(1A) receptor number. 5. Our data indicate that in serotonergic cell body areas the regulation of presynaptic 5-HT(1A) receptor function following repeated agonist administration occurs at the level of receptor-G protein interaction. In forebrain areas, however, the regulation of postsynaptic 5-HT(1A) receptor sensitivity appears not to be at the level of receptor-G protein coupling.


Subject(s)
Guanosine 5'-O-(3-Thiotriphosphate)/metabolism , Receptors, Serotonin/metabolism , Serotonin Receptor Agonists/pharmacology , 8-Hydroxy-2-(di-n-propylamino)tetralin/pharmacology , Animals , Autoradiography , GTP-Binding Proteins/metabolism , Image Processing, Computer-Assisted , Male , Piperazines/pharmacology , Pyridines/pharmacology , Raphe Nuclei/drug effects , Raphe Nuclei/metabolism , Rats , Rats, Sprague-Dawley , Receptors, Serotonin/drug effects , Receptors, Serotonin, 5-HT1 , Synapses/drug effects , Synapses/metabolism
10.
Neuropharmacology ; 39(11): 1996-2005, 2000 Aug 23.
Article in English | MEDLINE | ID: mdl-10963743

ABSTRACT

In the present study we have characterized the time course of effect of administration of the serotonin(2) (5-HT(2)) receptor antagonist mianserin, or the 5-HT(2) receptor agonist (+/-)-2,5-dimethoxy-4-iodophenyl-2-aminopropane (DOI), on 5-HT(2A) receptor binding sites and mRNA levels in rat frontal cortex. Radioligand binding and ribonuclease protection assays were performed with separate hemispheres of frontal cortex from each animal to examine concomitant changes in 5-HT(2A) receptor sites and mRNA levels. The decrease in cortical 5-HT(2A) receptor sites in response to chronic DOI administration was not accompanied by changes in 5-HT(2A) receptor mRNA. A single injection of DOI produced a transient decrease in 5-HT(2A) receptor mRNA levels detected 1 h post-injection. The density of 5-HT(2A) receptor sites, however, was not significantly reduced following a single injection of DOI. The down-regulation of cortical 5-HT(2A) receptor sites in response to a single injection of mianserin was accompanied by reductions in 5-HT(2A) receptor mRNA levels. Following 4 days of mianserin administration, however, we did not observe a change in 5-HT(2A) receptor mRNA levels, although 5-HT(2A) receptor density was decreased. Thus, changes in receptor mRNA may initially contribute to the down-regulation of 5-HT(2A) receptors in response to acute mianserin administration. Sustained changes in 5-HT(2A) receptor mRNA, however, appear not to be involved in maintaining the down-regulation of 5-HT(2A) receptor number with chronic mianserin administration. Mechanisms other than the regulation of receptor mRNA levels appear to underlie the down-regulation of 5-HT(2A) receptor sites in response to chronic administration of the agonist DOI.


Subject(s)
Amphetamines/pharmacology , Frontal Lobe/drug effects , Mianserin/pharmacology , Receptors, Serotonin/drug effects , Serotonin Antagonists/pharmacology , Serotonin Receptor Agonists/pharmacology , Animals , Frontal Lobe/metabolism , Ketanserin/pharmacology , Male , RNA, Messenger/drug effects , RNA, Messenger/metabolism , Rats , Rats, Sprague-Dawley , Receptor, Serotonin, 5-HT2A , Receptors, Serotonin/metabolism
11.
JAMA ; 284(8): 963-71, 2000.
Article in English | MEDLINE | ID: mdl-10944641

ABSTRACT

CONTEXT: Early-onset alcoholism differs from late-onset alcoholism by its association with greater serotonergic abnormality and antisocial behaviors. Thus, individuals with early-onset alcoholism may be responsive to treatment with a selective serotonergic agent. OBJECTIVE: To test the hypothesis that drinking outcomes associated with early vs late-onset alcoholism are differentially improved by the selective 5-HT(3) (serotonin) antagonist ondansetron. DESIGN: Double-blind, randomized, placebo-controlled clinical trial. SETTINGS: University of Texas Health Science Center in Houston (April 1995-June 1998) and University of Texas Health Science Center in San Antonio (July 1998-December 1999). PARTICIPANTS: A total of 321 patients with diagnosed alcoholism (mean age, 40.6 years; 70.5% male; 78.6% white) were enrolled, 271 of whom proceeded to randomization. INTERVENTIONS: After 1 lead-in week of single-blind placebo, patients were randomly assigned to receive 11 weeks of treatment with ondansetron, 1 microg/kg (n = 67), 4 microg/kg (n = 77), or 16 microg/kg (n = 71) twice per day; or identical placebo (n = 56). All patients also participated in weekly standardized group cognitive behavioral therapy. MAIN OUTCOME MEASURES: Self-reported alcohol consumption (drinks per day, drinks per drinking day, percentage of days abstinent, and total days abstinent per study week); and plasma carbohydrate deficient transferrin (CDT) level, an objective and sensitive marker of transient alcohol consumption. RESULTS: Patients with early-onset alcoholism who received ondansetron (1, 4, and 16 microg/kg twice per day) compared with those who were administered placebo, had fewer drinks per day (1.89, 1.56, and 1.87 vs 3.30; P =.03, P =.01, and P =.02, respectively) and drinks per drinking day (4.75, 4.28, and 5.18 vs 6.90; P =.03, P =.004, and P =.03, respectively). Ondansetron, 4 microg/kg twice per day, was superior to placebo in increasing percentage of days abstinent (70.10 vs 50.20; P =.02) and total days abstinent per study week (6.74 vs 5.92; P =.03). Among patients with early-onset alcoholism, there was a significant difference in the mean log CDT ratio between those who received ondansetron (1 and 4 microg/kg twice per day) compared with those who received the placebo (-0.17 and -0.19 vs 0.12; P =.03 and P =.01, respectively). CONCLUSION: Our results suggest that ondansetron (particularly the 4 microg/kg twice per day dosage) is an effective treatment for patients with early-onset alcoholism, presumably by ameliorating an underlying serotonergic abnormality. JAMA. 2000;284:963-971


Subject(s)
Alcoholism/prevention & control , Ondansetron/therapeutic use , Serotonin Antagonists/therapeutic use , Transferrin/analogs & derivatives , Adult , Alcoholism/blood , Analysis of Variance , Cognitive Behavioral Therapy , Dose-Response Relationship, Drug , Double-Blind Method , Female , Humans , Male , Middle Aged , Ondansetron/administration & dosage , Serotonin Antagonists/administration & dosage , Transferrin/metabolism
13.
Pharmacol Biochem Behav ; 63(2): 279-84, 1999 Jun.
Article in English | MEDLINE | ID: mdl-10371657

ABSTRACT

We have previously observed that intracerebroventricular infusion of a 5-HT2A receptor antisense oligonucleotide for 8 days results in an increase in cortical 5-HT2A receptor sites and an increase in central 5-HT2A receptor function as measured by quantitation of 5-HT2A receptor-mediated headshake behavior (28). Because lesioning serotonergic neurons or chronic administration of 5-HT2A receptor antagonists does not result in an increase in 5-HT2A receptor density or function in the brain, we have taken advantage of this unique upregulation of 5-HT2A receptors following 5-HT2A receptor antisense oligonucleotide infusion to study the modulation of D1 receptor-mediated behaviors by 5-HT2A receptors. Grooming behavior, elicited by acute injection of SKF 38393, was attenuated after chronic ICV infusion of a 5-HT2A receptor antisense oligonucleotide. There was also a reduction in vacuous chewing behavior induced by SKF 38393, which did not reach statistical significance. Other oral behaviors (i.e., tongue protrusions and gnawing at the cage bottom) were not attenuated. An increase in the density of cortical, as well as striatal 5-HT2A receptor sites was observed following chronic antisense oligonucleotide administration. There was no change in striatal D1 dopamine receptors following 5-HT2A receptor antisense oligonucleotide administration. SKF 38393-induced grooming behavior was also attenuated in naive rats pretreated acutely with the 5-HT2 receptor agonist DOI. These results suggest a role for the 5-HT2A receptor in the modulation of D1 receptor function.


Subject(s)
Grooming/physiology , Receptors, Dopamine D1/physiology , Receptors, Serotonin/drug effects , 2,3,4,5-Tetrahydro-7,8-dihydroxy-1-phenyl-1H-3-benzazepine/pharmacology , Animals , Base Sequence , Corpus Striatum/drug effects , Corpus Striatum/physiology , Dopamine Agonists/pharmacology , Grooming/drug effects , Ketanserin/metabolism , Male , Oligodeoxyribonucleotides, Antisense/pharmacology , Rats , Rats, Sprague-Dawley , Receptor, Serotonin, 5-HT2A , Receptors, Serotonin/genetics , Receptors, Serotonin/physiology , Serotonin Antagonists/metabolism
14.
Neuropsychopharmacology ; 19(5): 354-64, 1998 Nov.
Article in English | MEDLINE | ID: mdl-9778658

ABSTRACT

We have investigated the effect of 5-HT2 receptor agonist or antagonist administration on postsynaptic 5-HT1A receptor sensitivity assessed by two behavioral measures, reciprocal forepaw treading or hypothermia induced by acute injection of the 5-HT1A receptor agonist 8-OH-DPAT. The effectiveness of these drug treatments to downregulate 5-HT2A receptors was confirmed by measuring the binding of [3H]-ketanserin in cortical homogenates, because all of these drug treatments have been shown to result in the downregulation of 5-HT2A receptor sites. Acute or chronic treatment of rats with the 5-HT2 receptor antagonist mianserin, or chronic administration of the 5-HT2A receptor antagonist ketanserin, did not alter 8-OH-DPAT-induced hypothermia or forepaw treading. These data indicate that downregulation of 5-HT2A receptors is not sufficient to alter these postsynaptic 5-HT1A receptor-mediated responses. Chronic treatment of rats with the 5-HT2 receptor agonist DOI, however, resulted in the attenuation of both 5-HT1A receptor-mediated responses measured in separate experimental groups. The apparent desensitization of 5-HT1A receptors following chronic DOI treatment was not accompanied by a change in either the number or affinity of 5-HT1A receptor sites as measured by the binding of [3H]-8-OH-DPAT in hippocampal homogenates. Chronic activation of 5-HT2 receptors may be one mechanism by which the sensitivity postsynaptic 5-HT1A receptors can be regulated.


Subject(s)
Cerebral Cortex/drug effects , Receptors, Serotonin/drug effects , Receptors, Serotonin/physiology , Serotonin Agents/pharmacology , 8-Hydroxy-2-(di-n-propylamino)tetralin/pharmacology , Animals , Cerebral Cortex/physiology , Hypothermia/chemically induced , Hypothermia/physiopathology , Ketanserin/metabolism , Male , Motor Activity/drug effects , Motor Activity/physiology , Rats , Rats, Sprague-Dawley , Receptors, Serotonin, 5-HT1
15.
J Neurochem ; 71(4): 1457-63, 1998 Oct.
Article in English | MEDLINE | ID: mdl-9751178

ABSTRACT

The regulation of 5-HT2A receptor expression by an antisense oligodeoxynucleotide, complementary to the coding region of rat 5-HT2A receptor mRNA, was examined in a cortically derived cell line and in rat brain. Treatment of A1A1 variant cells, which express the 5-HT2A receptor coupled to the stimulation of phosphatidylinositol (PI) hydrolysis, with antisense oligodeoxynucleotide decreased the maximal stimulation of PI hydrolysis by the partial agonist quipazine and the number of 5-HT2A receptor sites as measured by the binding of 2-[125I]-iodolysergic acid diethylamide. Treatment of cells with random, sense, or mismatch oligodeoxynucleotide did not alter the stimulation of PI hydrolysis by quipazine or 5-HT2A receptor number. Intracerebroventricular infusion of antisense, but not mismatch, oligodeoxynucleotide for 8 days resulted in a significant increase in cortical 5-HT2A receptor density and an increase in headshake behavior induced by the 5-HT2 receptor agonist 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane. The density of cortical 5-HT2A receptors was not altered by administration of antisense oligodeoxynucleotide for 1, 2, or 4 days. We hypothesize that in brain this antisense oligodeoxynucleotide relieved some form of translational suppression, resulting in an increase in 5-HT2A receptor expression.


Subject(s)
DNA, Antisense/pharmacology , Receptors, Serotonin/biosynthesis , Receptors, Serotonin/drug effects , Animals , Cell Line , Cerebral Cortex/drug effects , Cerebral Cortex/metabolism , DNA, Antisense/administration & dosage , DNA, Antisense/genetics , Injections, Intraventricular , Male , Rats , Rats, Sprague-Dawley , Receptor, Serotonin, 5-HT2A , Receptors, Serotonin/genetics
16.
J Pharmacol Exp Ther ; 278(3): 1138-45, 1996 Sep.
Article in English | MEDLINE | ID: mdl-8819496

ABSTRACT

P11 cells were transfected with DNA for the human 5-hydroxytryptamine1A (5-HT1A) receptor. These cells stably expressed the 5-HT1A receptor coupled to the inhibition of adenylyl cyclase, and not to the stimulation of phosphoinositide hydrolysis. Homologous and heterologous regulation of the 5-HT1A receptor was studied in this cell system. Pretreatment of P11-5HT1A cells with the 5-HT1 receptor agonist 5-carbox-amidotryptamine (5-CT) resulted in a 3-fold increase in both basal and forskolin-stimulated cAMP accumulation, and desensitization of the 5-HT1A receptor as indicated by a decrease in the potency of 8-hydroxydipropylaminotetralin (8-OH-DPAT) to inhibit forskolin-stimulated cAMP accumulation (vehicle-treated cells: EC50 = 2.3 +/- 0.8 nM; 5-CT-treated cells: 9.9 +/- 0.4 nM). The sensitization of adenylyl cyclase as a result of chronic agonist exposure was prevented by the 5-HT1A antagonist WAY100635, which indicated that the effect of 5-CT pretreatment on basal and forskolin-stimulated cAMP accumulation was mediated by 5-HT1A receptor activation. Pretreatment of cells with pertussis toxin abolished the inhibition of forskolin-stimulated cAMP accumulation mediated by 5-HT1A receptor activation and prevented the sensitization of adenylyl cyclase as a result of chronic 5-HT1A receptor agonist exposure. Pretreatment of P11-5HT1A cells with the phorbol ester, phorbol 12-myristate 13-acetate (PMA), also resulted in desensitization of the 5-HT1A receptor, as indicated by a marked decrease in the potency and intrinsic activity of 8-OH-DPAT. No change in the binding characteristics (i.e., Kd or Bmax) of [3H]8-OH-DPAT to 5-HT1A receptor sites was observed after 5-CT or PMA treatments. Activation of alpha-1 adrenergic receptors, but not 5-HT2A receptors, had effects on 5-HT1A receptor responsiveness similar to those seen with PMA pretreatment. In P11-5HT1A cells, homologous regulation of the 5-HT1A receptor was characterized by sensitization of adenylyl cyclase and a decrease in agonist potency, whereas heterologous regulation of the 5-HT1A receptor was characterized by a greater decrease in agonist potency, as well as a marked decrease in intrinsic activity.


Subject(s)
Receptors, Serotonin/physiology , 8-Hydroxy-2-(di-n-propylamino)tetralin/metabolism , Adenylyl Cyclases/metabolism , Animals , Cell Line , Humans , Phosphatidylinositols/metabolism , Protein Kinase C/physiology , Rats , Receptors, Serotonin, 5-HT1 , Second Messenger Systems , Serotonin/analogs & derivatives , Serotonin/pharmacology , Serotonin Receptor Agonists/pharmacology , Signal Transduction , Tetradecanoylphorbol Acetate/pharmacology , Transfection
18.
Synapse ; 17(1): 1-15, 1994 May.
Article in English | MEDLINE | ID: mdl-8042142

ABSTRACT

The binding of 3H-CN-IMI to 5-HT uptake sites, as measured by quantitative autoradiography, was used as a marker of serotonergic neurons. Within the dorsal raphe nucleus the binding of 3H-CN-IMI was compared in adjacent coronal sections of rat brain to the binding of 3H-DPAT to 5-HT1A receptors, which have a known somatodendritic localization. The heterogeneous pattern of binding of these two radioligands within the dorsal raphe nucleus was similar and corresponded to the distribution of serotonergic cell bodies as visualized by 5-HT immunohistochemistry. Intracerebroventricular administration of 5,7-dihydroxytryptamine (5,7-DHT), which caused a dramatic loss of 5-HT immunoreactivity and 3H-DPAT binding to 5-HT1A receptors, resulted in a marked reduction of 3H-CN-IMI binding in this nucleus. Treatment of rats with a dose of para-chloroamphetamine (PCA) which has been reported to selectively lesion serotonergic processes arising from the dorsal raphe nucleus, while sparing serotonergic cell bodies and projections from the median raphe nucleus, did not alter the binding of 3H-DPAT or 3H-CN-IMI in the dorsal raphe nucleus; serotonergic cell bodies appeared morphologically unaffected. The lack of effect of PCA treatment on the binding of 3H-DPAT and 3H-CN-IMI is consistent with a somatodendritic localization of the 5-HT transporter in the dorsal raphe nucleus. PCA treatment appeared to produce a moderate loss of serotonergic innervation in serotonergic terminal field areas as visualized by serotonin immunohistochemistry. The reductions in 3H-CN-IMI binding observed in terminal field areas (24 to 69%) following treatment of rats with PCA did not reflect a marked differential innervation of forebrain areas by the dorsal and medial raphe nuclei as expected from previous biochemical studies, and were not entirely consistent with the findings of neuroanatomical studies using histochemical techniques. Site-specific injection of 5,7-DHT into the dorsal raphe nucleus produced an 80 +/- 11% reduction in the binding of 3H-CN-IMI in this nucleus, whereas the binding of 3H-CN-IMI in the median raphe nucleus was not reduced.(ABSTRACT TRUNCATED AT 400 WORDS)


Subject(s)
Carrier Proteins/metabolism , Membrane Glycoproteins/metabolism , Membrane Transport Proteins , Nerve Tissue Proteins , Raphe Nuclei/physiology , Serotonin/physiology , Synaptic Transmission , 5,7-Dihydroxytryptamine/pharmacokinetics , Animals , Autoradiography , Imipramine/analogs & derivatives , Imipramine/pharmacokinetics , Male , Raphe Nuclei/drug effects , Raphe Nuclei/metabolism , Rats , Rats, Sprague-Dawley , Serotonin/metabolism , Serotonin Antagonists/pharmacokinetics , Serotonin Plasma Membrane Transport Proteins , Tetrahydronaphthalenes/pharmacokinetics , p-Chloroamphetamine/pharmacokinetics
19.
J Pharmacol Exp Ther ; 268(2): 935-42, 1994 Feb.
Article in English | MEDLINE | ID: mdl-8114008

ABSTRACT

[125I](R,S)-trans-7-Hydroxy-2-[N-propyl-N-(3'-iodo-2'-propenyl)- amino]tetralin ([125](R,S)-trans-7-OH-PIPAT) has been shown to bind with high affinity to dopamine D3 receptors expressed in Spodoptera frugiperda cells. No specific binding was seen in Spodoptera frugiperda cells expressing a high density of D2 receptors. It was therefore, suggested that [125I] (R,S)-trans-7-OH-PIPAT selectively labels D3 receptors. In the present study, saturation binding of [125I](R)-trans-7-OH-PIPAT to membranes from rat olfactory tubercle resulted in markedly curvilinear Scatchard plots, suggesting that the radioligand was binding to more than one receptor class or affinity state. [125I] (R)-trans-7-OH-PIPAT bound with high affinity to membranes from human embryonic kidney-293 cells expressing transfected D2 or D3 receptors and to membranes from Chinese hamster ovary cells expressing serotonin1A receptors. Binding of [125I](R)-trans-7-OH-PIPAT to serotonin1A and D2 receptors was decreased or eliminated in the presence of NaCl and/or guanylyl-imidodiphosphate [Gpp(NH)p]. In the presence of Gpp(NH)p and NaCl, a linear Scatchard plot with a Kd value of 0.4 nM and a density of 100 fmol/mg of protein was obtained in membranes from rat olfactory tubercle. Agonists and antagonists inhibited binding of [125I](R)-trans-7-OH-PIPAT with a rank order of potency consistent with an interaction at D3 receptors. These results suggest that, in the presence of Gpp(NH)p and NaCl, [125I](R)-trans-7-OH-PIPAT specifically labels D3 receptors.


Subject(s)
Olfactory Pathways/metabolism , Receptors, Dopamine/metabolism , Tetrahydronaphthalenes/metabolism , Animals , Cell Line , Guanylyl Imidodiphosphate/pharmacology , Humans , Rats , Rats, Sprague-Dawley , Receptors, Dopamine/genetics , Receptors, Dopamine D2/metabolism , Receptors, Dopamine D3 , Receptors, Serotonin/metabolism , Sodium Chloride/pharmacology , Transfection
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