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In the central nervous system, nitric oxide (NO), a free gas with multitudinous bioactivities, is mainly produced from the oxidation of L-arginine by neuronal nitric oxide synthase (nNOS). In the past 20 years, the studies in our group and other laboratories have suggested a significant involvement of nNOS in a variety of neurological and neuropsychiatric disorders. In particular, the interactions between the PDZ domain of nNOS and its adaptor proteins, including post-synaptic density 95, the carboxy-terminal PDZ ligand of nNOS, and the serotonin transporter, significantly influence the subcellular localization and functions of nNOS in the brain. The nNOS-mediated protein-protein interactions provide new attractive targets and guide the discovery of therapeutic drugs for neurological and neuropsychiatric disorders. Here, we summarize the work on the roles of nNOS and its association with multiple adaptor proteins on neurological and neuropsychiatric disorders.
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Humanos , Óxido Nítrico Sintase Tipo I/metabolismo , Proteínas Adaptadoras de Transdução de Sinal , Encéfalo/metabolismo , Doenças do Sistema NervosoRESUMO
In the central nervous system, nitric oxide (NO), a free gas with multitudinous bioactivities, is mainly produced from the oxidation of L-arginine by neuronal nitric oxide synthase (nNOS). In the past 20 years, the studies in our group and other laboratories have suggested a significant involvement of nNOS in a variety of neurological and neuropsychiatric disorders. In particular, the interactions between the PDZ domain of nNOS and its adaptor proteins, including post-synaptic density 95, the carboxy-terminal PDZ ligand of nNOS, and the serotonin transporter, significantly influence the subcellular localization and functions of nNOS in the brain. The nNOS-mediated protein-protein interactions provide new attractive targets and guide the discovery of therapeutic drugs for neurological and neuropsychiatric disorders. Here, we summarize the work on the roles of nNOS and its association with multiple adaptor proteins on neurological and neuropsychiatric disorders.
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Resumen Introducción: Los cardiomiocitos poseen la maquinaria bioquímica capaz de sintetizar, utilizar y recapturar serotonina. Objetivo: Determinar si la miocardiopatía hipertrófica (MCH) induce cambios en la expresión de la triptófano-5-hidroxilasa (TPH) 1 y 2, el transportador de serotonina (SERT) y los receptores serotoninérgicos (RS). Métodos: Estudio transversal de cinco bloques de tejido de corazones con MCH y cinco bloques de corazones de control. Se obtuvieron cinco cortes de la pared libre del ventrículo izquierdo (PLVI) y del septum interventricular (SIV) de cada bloque, para determinar la expresión de TPH1 y TPH2, SERT y RS con anticuerpos por inmunofluorescencia. La inmunofluorescencia fue evaluada mediante t de WELCH, con nivel de significación de p < 0.05. Resultados: La PLVI y el SIV de los corazones con MCH mostraron aumento de la expresión de TPH1 y TPH2, así como de los receptores 5-HT2A y 5-HT2B en comparación con los controles (p < 0.01). El receptor 5-HT4 y SERT aumentaron en el SIV de los corazones con MCH (p < 0.01). Conclusiones: Se demostró aumento de las expresiones de TPH, SERT y RS en los cardiomiocitos de los corazones con MCH en comparación con los controles, lo cual podría participar en la fisiopatología de la MCH en los humanos.
Abstract Introduction: Cardiomyocytes have a biochemical machinery with the capacity to synthesize, utilize and reuptake serotonin. Objective: To determine whether hypertrophic cardiomyopathy (HCM) induces changes in the expression of tryptophan-5-hydroxylase (TPH) 1 and 2, serotonin transporter (SERT) and serotonergic receptors (SR). Methods: Cross-sectional study of five tissue blocks from hearts with HCM and five controls. Five sections of the left ventricular free wall (LVFW) and interventricular septum (IVS) were obtained from each block to determine the expression of TPH1 and TPH2, SERT and SRs by immunofluorescence with specific antibodies. Immunofluorescence was evaluated by WELCH t-test, with a level of significance of p < 0.05. Results: LVFW and IVS of hearts with HCM showed an increase in the expression of TPH1 and TPH 2 and 5-HT2A and 5-HT2B receptors in comparison with controls (p < 0.01). The 5-HT4 receptor and SERT showed an increase in the IVS of hearts with HCM (p < 0.01). Conclusions: This study demonstrated an increased expression of TPH, SERT and SRs in cardiomyocytes from hearts with HCM in comparison with controls, which could be involved in the pathophysiology of HCM in humans.
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Aim To study the protective effect of fluoxetine hydrochloride on brain tissues of rats with simulated high altitude cerebral edema(HACE)and its mechanism.Methods The optimal dosage and time of fluoxetine hydrochloride were determined by the hypoxia tolerance test of mice under normal pressure.The rat model of brain edema at high altitude was established by large-scale low-pressure oxygen chamber.HE staining was used to observe the pathological changes of brain tissues in rats.Microplate reader was used to detect the corresponding indexes of oxidative stress such as malondialdehyde(MDA)level and superoxide dismutase(SOD)activity.The expressions of hypoxia-related proteins HIF-1α,VEGF,MMP-9,AQP4 and SERT were detected by Western blot.Results Compared with the hypoxia model group,after the intervention of fluoxetine hydrochloride,the survival time of mice was prolonged,and the middle dose of fluoxetine(14 mg·kg-1)had the best effect,with an extension rate of 17.78%.The pathological damage of brain was improved,the water content of brain decreased,and the permeability of blood-brain barrier decreased.MDA content in rat brain decreased and SOD activity increased.Western blot results showed that HIF-1α,VEGF,MMP-9,AQP4,SERT protein were significantly down-regulated.Conclusions Fluoxetine has protective effect on rats with brain edema at high altitude,and its mechanism may be related to improving oxidative stress,activating HIF-1α/VEGF/MMP-9 signaling pathway and affecting the expression of SERT protein.SERT may be a potential target for treating brain edema at high altitude.
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Objective:To observe the effect of Tongxie Yaofang on the expressions of colon serotonin transporter （SERT）， liver 5-hydroxytryptamine<sub>2A</sub> receptor （5-HT<sub>2A</sub>R） protein， serum 5-HT and inflammatory factors in ulcerative colitis （UC） model rats of liver stagnation and spleen deficiency， in order to explore the basis of syndrome of liver stagnation and spleen deficiency and the intervention mechanism of Tongxie Yaofang. Method:Fifty male SD rats were randomly divided into blank control group， model group， high， medium and low-dose Tongxie Yaofang group （10，5，2.5 g·kg<sup>-1</sup>）， and salazosulacil group （0.3 g·kg<sup>-1</sup>）. The ulcerative colitis model of liver depression and spleen deficiency was established by 2，4，6-trinitrobenzene sulfonic acid （TNBS）/ethanol solution enema + restraint stress + diet loss. After successful modeling， the samples were collected after 21 days of drug intervention. Htoxylin eosin （HE） staining and oil red staining were used to observe the pathological changes of colon and liver in each group. Serum interleukin-6 （IL-6）， IL-9， 5-HT and superoxide dismutase （SOD） were detected by enzyme linked immunosorbent assay （ELISA）. Protein expressions of SERT in the colons and 5-HT<sub>2A</sub>R in liver of rats were detected by Western blot. Result:Compared with the normal group， obvious ulcers were formed in the colon and lipid droplets in the liver increased in the model group， serum levels of IL-6， IL-9 and 5-HT in the model group increased， while the level of SOD decreased （<italic>P</italic><0.05）. The protein expression of SERT in colon decreased， whereas the protein expression of 5-HT<sub>2A</sub>R in liver increased （<italic>P</italic><0.05）. Compare with model group， the pathological damage of colon was improved， and the formation of lipid droplets in liver was reduced in high， medium-dose Tongxie Yaofang groups and sulfasalazine group. The serum levels of IL-6， IL-9 and 5-HT decreased， while the level of SOD increased in Tongxie Yaofang group and sulfasalazine group （<italic>P</italic><0.05）. The protein expression of SERT in colon increased in high，low-dose Tongxie Yaofang groups and sulfasalazine group， and the protein expression of 5-HT<sub>2A</sub>R in liver decreased in medium， low dose Tongxie Yaofang groups and sulfasalazine group （<italic>P</italic><0.05）. Conclusion:Tongxie Yaofang may reduce the content of 5-HT， and regulate the intestinal motility and sensory system by up-regulating the expression of SERT in the colon， inhibit the expressions of IL-6，IL-9 and other inflammatory factors， and play an anti-inflammatory role， reduce the content of 5-HT and the expression of 5-HT<sub>2A</sub>R in the liver， increase the level of SOD， regulate emotion and lipid metabolism in the liver， and then exert the intervention effect on ulcerative colitis with liver depression and spleen deficiency on the whole.
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Objective • To discuss the effects of citalopram on miRNA-16/serotonin transporter (SERT) pathway in peripheral blood of the patients with depression. Methods • Forty-five patients with depression without medication (untreated group), 32 patients with depression treated with medicine(drug treated group) and 32 healthy people (control group) were enrolled in the study. Hamilton Depression Scale-17 items were used to evaluate the depressive symptoms. The expression level of plasma miRNA-16 was detected by fluorescence quantitative PCR, and the level of SERT protein in platelets was detected by Western blotting. Fourteen of the baseline patients who were treated with citalopram were followed up for 2 months. After the follow-up, the evaluation of HAMD-17, the detection of miRNA-16 and SERT protein were conducted. Results • There was no significant difference in the expression level of plasma miRNA-16 among the three groups (F=0.421, P=0.657). There was no significant difference of SERT protein expression in the platelets among the three groups (F=0.112, P=0.894). The follow-up study showed that the HAMD-17 score decreased after 2 months (Z=.3.187, P=0.001), the average expression level of plasma miRNA-16 increased (t=2.455, P=0.032), and the expression of SERT protein in the platelets did not change (t=.0.750, P=0.470) in 14 patients who were treated with citalopram. Conclusion • Citalopram, a serotonin reuptake inhibitor, can down-regulate the expression of plasma miRNA-16 in patients with depression, and the decrease of the platelet serotonin is not caused by the decrease of SERT protein on platelet membrane, but may be related to the decrease of the SERT function.
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Objective: The current research focuses on neurotransmitter serotonin levels and the work was planned to study the comparison between children (female and male) as well as the special need children (with abnormal behaviors ADHD). Materials and methods: Total 86 children from certain primary school were selected, 56 of them were with ADHD (27 boys and 29 girls were present) and 30 healthy individuals (16 boy and 14 girls). ADHD was diagnosed by certain questionnaire form prepared for this study, blood samples were gathered, and were send for serotonin examination which was achieved with a certain enzyme linked immune sorbent assay (ELISA). Results were analyzed using special statistic program (SPSS version 18). Results: The collected data indicate that serotonin level was high (121.70 ± 4.05 ng/ml) in ADHD children compared with normal children (85.64 ± 2.43 ng/ml), and highly significant decrease in level of serotonin transporter in ADHD than in control samples (9.87 ± 0.29; 13.17 ± 0.50 ng/ml correspondingly). Conclusion: The increase and decrease in the level of serotonin serum and serotonin transporters could be related to the environmental issues that contribute to the disruption of behavior particularly rise in poor security and hot regions.
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Aim To investigate the effect of Radices Paeoniae Alba extract on estrogen receptor (3 ( ERp ) and serotonin transporter(SERT) in the hippocampus of rat model of premenstrual syndrome( PMS) liver-qi inverse syndrome. Methods The rats were randomly divided into the normal group, the model group, the Radices Paeoniae Alba extract group, the fluoxetine group and the ER(3 agonist group by the vaginal smear microscopy and the open field score. The model group rats were prepared by electrical stimulation to prepare PMS liver-qi inverse syndrome. After five days of administration, the hippocampal tissues of each group were separated. ERp and SERT mRNA and protein expression in each group were determined by RT-PCR and immunofluorescence histochemical method. Results During the estrous reception period, there was no significant difference in the behavior score of the open- field test and the expression of ERp and SERT mRNA and protein in the hippocampus of each group. Estrous non-reception period; compared with normal rats, the total distance of open field test in model rats increased significantly. The expression of ER(3 mRNA in hippocampus significantly decreased and SERT mRNA significantly increased. In model group, the pyramidal cells in hippocampal CA1 and CA3 brain areas of rats were irregularly arranged, and the number of ERp positive cells and protein expression decreased. The number of SERT positive cells and protein expression were significantly up-regulated. After the Radices Paeoniae Alba extract was given, the above indexes were significantly improved, and no significant difference was found between them and fluoxetine group and ER agitator group. Conclusions Radices Paeoniae Alba extract may adjust the PMS card of diseases with inverse nega-tive emotions possibly by improving the hippocampus ER|3 and SERT mRNA and protein expression.
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BACKGROUND/AIMS: MicroRNAs (miRNAs) were reported to be responsible for intestinal permeability in diarrhea-predominant irritable bowel syndrome (IBS-D) rats in our previous study. However, whether and how miRNAs regulate visceral hypersensitivity in IBS-D remains largely unknown. METHODS: We established the IBS-D rat model and evaluated it using the nociceptive visceral hypersensitivity test, myeloperoxidase activity assay, restraint stress-induced defecation, and electromyographic (EMG) activity. The distal colon was subjected to miRNA microarray analysis followed by isolation and culture of colonic epithelial cells (CECs). Bioinformatic analysis and further experiments, including dual luciferase assays, quantitative real-time polymerase chain reaction, western blot, and enzyme-linked immunosorbent assay, were used to detect the expression of miRNAs and how it regulates visceral hypersensitivity in IBS-D rats. RESULTS: The IBS-D rat model was successfully established. A total of 24 miRNAs were differentially expressed in the distal colon of IBS-D rats; 9 were upregulated and 15 were downregulated. Among them, the most significant upregulation was miR-200a, accompanied by downregulation of cannabinoid receptor 1 (CNR1) and serotonin transporter (SERT). MiR-200a mimic markedly inhibited the expression of CNR1/SERT. Bioinformatic analysis and luciferase assay confirmed that CNR1/SERT are direct targets of miR-200a. Rescue experiments that overexpressed CNR1/SERT significantly abolished the inhibitory effect of miR-200a on the IBS-D rats CECs. CONCLUSIONS: This study suggests that miR-200a could induce visceral hyperalgesia by targeting the downregulation of CNR1 and SERT, aggravating or leading to the development and progression of IBS-D. MiR-200a may be a regulator of visceral hypersensitivity, which provides potential targets for the treatment of IBS-D.
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Animais , Ratos , Western Blotting , Colo , Biologia Computacional , Defecação , Diarreia , Regulação para Baixo , Ensaio de Imunoadsorção Enzimática , Células Epiteliais , Hiperalgesia , Hipersensibilidade , Síndrome do Intestino Irritável , Luciferases , Análise em Microsséries , MicroRNAs , Modelos Animais , Permeabilidade , Peroxidase , Reação em Cadeia da Polimerase em Tempo Real , Receptores de Canabinoides , Proteínas da Membrana Plasmática de Transporte de Serotonina , Serotonina , Regulação para CimaRESUMO
This study is to investigate the effect of antidepressant medicine prescription Dingzhi Xiaowan (DZ) on miR-16 expression levels in vitro and in vivo, and to explore the mechanism of DZ elevated levels of 5-HT from the perspective of post transcriptional regulation. Firstly, a chronic unpredictable mild moderate stimulation (CUMS) combined with solitary rising depression rat model was established, the behavior changes were detected after different doses of DZ (600, 300, 150 mg·kg⁻¹) given for 3 weeks, high performance liquid chromatography (HPLC) was used to detect 5-HT level in hippocampal, PCR method was used to observe the effect of DZ on the expression of SERT mRNA and miR-16 in hippocampus of CUMS rat. The effects of DZ (10, 100, 200, 500 mg·L⁻¹) on the expression of miR-16 and SERT mRNA in the cell model induced by miR-16 silencing and corticosterone or glutamate injury were observed in primary cultured hippocampal neurons of rats in vitro. It was found that 300 mg·kg⁻¹ and 600 mg·kg⁻¹ DZ could significantly improve the behavioral score of CUMS rats, increase the level of 5-HT in hippocampus, and increase the expression of miR-16 and decrease the expression of SERT in the hippocampus of rats. At the same time, in primary cultured hippocampal neurons, 100, 200, 500 mg·L⁻¹ of DZ could significantly increase the expression level of miR-16 in miR-16 silencing and corticosterone or glutamate injury cell model, and decrease the expression level of SERT significantly. So DZ could inhibit the reuptake of 5-HT by inhibiting the expression of SERT by up regulating the expression level of miR-16, and finally increase the level of 5-HT in the brain and exert antidepressant effect.
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Objective To explore the influence of the lactation exposure to fluoxetine on offspring's behavior and serotonin transporter (SERT) and tryptophan hydroxylase (TPH). Methods Six SD pregnant rats were randomly divided into 2 groups (n=3 each group). Experimental maternal rats were intraperitoneally injected with fluoxetine at a dose of 12 mg/kg from postnatal day 5 to 21. The control group were injected with the same amount of normal saline. In infancy, the offspring's weight, hair length, eye opening and auditory development were measured. The free suspension test and bur?ied food pellets test were applied to evaluate the offspring's behaviors. After postnatal day 21, all the offspring were wean. At early childhood (P35d) and adulthood (P75d), 6 offspring rats from each group were executed to examine SERT and TPH in the prefrontal cortex by immunohistochemistry. Results The offspring's weight of experimental group was significantly lower than control group (P<0.05). The sensitivity of auditory in experimental group was significantly higher than control group (P<0.01). The time of free suspension in experimental group significantly was decreased comparing to control group (P<0.01). The SERT and TPH in prefrontal cortex was significantly lower in experimental group than those in control group either at childhood (P35d) or at adulthood (P75d) (P<0.05). Conclusion Lactation exposure to fluoxetine re?sults in offspring's abnormal development and behaviors through down-regulation of SERT and TPH in the prefrontal cor?tex.
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The SLC6A4 gene encodes the serotonin transporter SERT. Since the discovery of the role of SLC6A4 polymorphisms on human behavior, there is an increasingly growing wealth of information regarding SLC6A4 gene variants associated with anxiety and mood disorders, as well as their pharmacogenetic implications. In this brief review, the main discoveries on SLC6A4 variants, their functional impact and their suggested roles in neuropsychiatric and neurodevelopmental disorders are discussed.
El gen SLC6A4 codifica el transportador de serotonina SERT. Desde el descubrimiento inicial del rol que tienen polimorfismos de SLC6A4 en el comportamiento humano, hay una creciente cantidad de información acerca de variantes genéticas de SLC6A4 asociadas con trastornos de ansiedad y de estado de ánimo, así como de sus implicancias farmacogenéticas. En esta breve revisión, se discuten los principales descubrimientos de variantes de SLC6A4, su impacto funcional y sus roles sugeridos en enfermedades neuropsiquiátricas y de neurodesarrollo.
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Humanos , Serotonina , Proteínas da Membrana Plasmática de Transporte de Serotonina , Doenças do Sistema NervosoRESUMO
Aims: The aim of our study was to investigate the effects of continuous action of ultrasonic waves of variable frequencies on behavior of rats in "classical" tests used to reveal depression-like behavior, to evaluate the influence of different psychotropic drugs on rates of these tests and to analyze expression of several genes involved in pathogenesis of depression. Study Design: Rats in individual cages were exposed to ultrasonic irradiation for 21 days. Place and Duration of Study: V.P. Serbsky National Recearh Center For Social and Forensic Psychiatry, Department of Basic and Applied Neurobiology, Moscow, Russian Federation, between November 2012 and January 2013. Methodology: 48male non-pedigree albino rats were divided into 5 groups: nonultrasound- saline, ultrasound-saline, ultrasound-fluoxetine, ultrasound-bupropion and ultrasound-tianeptine. After 21 days of irradiation social interaction test, forced swimming test and sucrose preference test (anhedonia test) were conducted. Than rats were decapitated and prefrontal cortex were taken for RT-qPCR gene expression analysis of 5-HT1A, 5-HT2A, 5HT1B, 5HT2B receptors and SERT. Results: Depression-like behavior manifests itself in reduced social activity in social interaction test, increased immobility in forced swimming test and lower sucrose consumption in anhedonia test. The administrated antidepressants demonstrated their effectiveness, except for bupropion in the social interaction test. RT-qPCR gene expression analysis showed reduced expression of 5HT2A receptor gene and increased expression of SERT gene in the prefrontal cortex of rats stressed with ultrasonic radiation. Conclusion: The obtained data allow to conclude that further investigations with lager number of animals, extended tests battery may allow to claim that this model meets the main requirements set to animal models (face, predictive and construct validity) and can be used in studies of depression-like disorders caused by a situation of informational uncertainty and in pre-clinical development of new antidepressants.
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Visceral hypersensitivity plays an important role in motor and sensory abnormalities associated with irritable bowel syndrome, but the underlying mechanisms are not fully understood. The present study was designed to evaluate the expression of the 5-HT4 receptor and the serotonin transporter (SERT) as well as their roles in chronic visceral hypersensitivity using a rat model. Neonatal male Sprague-Dawley rats received intracolonic injections of 0.5% acetic acid (0.3-0.5 mL at different times) between postnatal days 8 and 21 to establish an animal model of visceral hypersensitivity. On day 43, the threshold intensity for a visually identifiable contraction of the abdominal wall and body arching were recorded during rectal distention. Histological evaluation and the myeloperoxidase activity assay were performed to determine the severity of inflammation. The 5-HT4 receptor and SERT expression of the ascending colon were monitored using immunohistochemistry and Western blot analyses; the plasma 5-HT levels were measured using an ELISA method. As expected, transient colonic irritation at the neonatal stage led to visceral hypersensitivity, but no mucosal inflammation was later detected during adulthood. Using this model, we found reduced SERT expression (0.298 ± 0.038 vs 0.634 ± 0.200, P < 0.05) and increased 5-HT4 receptor expression (0.308 ± 0.017 vs 0.298 ± 0.021, P < 0.05). Treatment with fluoxetine (10 mg·kg-1·day-1, days 36-42), tegaserod (1 mg·kg-1·day-1, day 43), or the combination of both, reduced visceral hypersensitivity and plasma 5-HT levels. Fluoxetine treatment increased 5-HT4 receptor expression (0.322 ± 0.020 vs 0.308 ± 0.017, P < 0.01) but not SERT expression (0.219 ± 0.039 vs 0.298 ± 0.038, P = 0.654). These results indicate that both the 5-HT4 receptor and SERT play a role in the pathogenesis of visceral hypersensitivity, and its mechanism may be involved in the local 5-HT level.
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Animais , Masculino , Ratos , Hipersensibilidade/metabolismo , Síndrome do Intestino Irritável/metabolismo , /metabolismo , Proteínas da Membrana Plasmática de Transporte de Serotonina/metabolismo , Vísceras/metabolismo , Animais Recém-Nascidos , Western Blotting , Doença Crônica , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Fluoxetina/farmacologia , Hipersensibilidade/tratamento farmacológico , Imuno-Histoquímica , Síndrome do Intestino Irritável/induzido quimicamente , Síndrome do Intestino Irritável/tratamento farmacológico , Ratos Sprague-Dawley , Índice de Gravidade de Doença , Inibidores Seletivos de Recaptação de Serotonina/farmacologiaRESUMO
The serotonin transporter (SERT) is the target site for serotonin reuptake inhibitors, which are the most widely used agents for treating various psychiatric diseases including depression. The SERT is a member of a large family of homologous integral membrane proteins. This transporter takes up 5-HT in a process that is coupled to the transmembrane movement of Na+, Cl-, and K+. The SERT may operate in at least two modes, an alternating access carrier or a channel. The function of SERT is acutely regulated by various protein kinases and phosphatases. The SERT gene is located on chromosome 17 and has several polymorphisms including 5-HTTLPR and intron 2 VNTR. Most studies involving the association between 5-HTTLPR and the response to SSRI in depression reported that l/l genotype showed better response and fewer side effects. But, it is too early to draw definite conclusion of the effects of 5-HTTLPR on anti-depressant treatment. Therefore, it is necessary to perform further studies reflecting various ethnicities and genetics of subjects as well as the environmental interactions. This review discusses recent advances in defining the structure, the action mechanism, the location, and the regulation of SERT. Furthermore, it discusses the function of SERT polymorphisms and its implications on the anti-depressant therapies.