Expression Of Serotonin Transporter Protein Research Articles

The effects of proinflammatory cytokines on expression of serotonin transporters in human enterocyte-like cell line

Objective To determine the effect of proinflammatory cytokines including interleukin-lbeta (IL-1β) and tumor necrosis factor-Alpha (TNF-α) on human serotonin transporter (SERT) in human enterocyte-like cell line Caco-2.Methods Caco2 cells were cultured for 5 days,then were divided into control group.IL-1β treated group (50 ng/ml) and TNF-α treated group (50ng/ml) for 2.24,48 and 72 hours.The expression of SERT mRNA was detected by RT-PCR at 2,24 and 48 hours and its protein expression was measured by Western blotting at 24,48 and 72 hours.Results The expression of SERT mRNA at 2,24 and 48 hours was lower both in IL-1β treated group (1.393±1.184,1.064±0.625 and 1.013±0.415,respectively) and TNF-α treated group (1.000±0.000,0.829±0.162and 0.945±0.147,respectively) in comparison with control group (2.282±1.367,1.586±0.421 and 1.86 ±O.496,respectively).There was significant difference among three groups (P<0.01).The expression of SERT protein at 24,48 and 72 hours was lower in IL-1β treated group and TNF-α treated group than in control group.Conclusions The IL-1β and TNF-α can inhibit the expressions of SERT mRNA and protein in Caco-2 cells,which indicates that IL-1β and TNF-α may change visceral sensitivity by influencing peripheral 5-hydroxytryptamine levels. Key words: Colonic neoplasms Interleukins; Tumor necrosis factor-Alpha; Serotonin transporter; Cell line,tumor

Serotonin transporter gene deficiency is associated with sudden death of newborn mice through activation of TGF-β1 signalling

The serotonin transporter (SERT) gene has been proposed as a candidate gene responsible for the sudden infant death syndrome (SIDS). In this study, for the first time we obtained a SERT-knockout (KO) mouse model which reproduces SIDS phenotype. SERT-KO mice were generated by mating SERTCre/+ heterozygous mice. The SERT-KO mouse embryos at the pre-natal stage E18.5 were lacking of SERT mRNA and protein expression in the heart. A premature death of 75% of SERT-KO mice occurred in the first week after birth. LacZ staining of whole mounts and tissue sections of the heart from SERTCre/+;ROSA26R adult mice and E18.5 embryos demonstrated a marked localized expression of SERT in the right ventricle, the conal region, the vasculature, the atrial septum, the ventricular valves, and the sinoatrial node of the conduction system. These data suggest a cardiac phenotype for the sudden death of SERT-KO mice. Histological analysis of heart sections showed that SERT-KO mice develop cardiac fibrosis. Increased collagen accumulation in the myocardium and the valvular and perivascular regions, and enhanced expression of α-smooth muscle actin were detected in the heart of SERT-KO mice versus wild-type (WT) mice. Interestingly, higher expression levels of the 5-HT2A receptor and increased levels of phospho-SMAD2/3 and phospho-ERK1/2 were detected in SERT-KO mouse heart versus WT mice. Overall, our findings provide i) new insights into the role of SERT gene in SIDS, and ii) the first in vivo validation of the molecular mechanism involving the activation of TGF-β1 signalling in the cardiac fibrosis.

Expression of placental serotonin transporter and 5-HT 2A receptor in normal and gestational diabetes mellitus pregnancies

Recent studies reveal that serotonin (5-hydroxytryptamine, 5-HT) might play a role in the aetiology of gestational diabetes mellitus (GDM). The involvement of the serotonin transporter (SERT) and 5-HT(2A) receptor (5-HT(2A)R) in diabetes has also been suggested. However, placental SERT and 5-HT(2A)R have never been studied in GDM-complicated pregnancies. The aim of this study was to investigate the effect of GDM on the expression of both placental SERT and 5-HT(2A)R. First, immunohistochemical analysis demonstrated the presence of SERT and 5-HT(2A)R proteins in the villous trophoblast and the fetal capillary endothelium of normal term placental tissue. Protein and mRNA expression of SERT and 5-HT(2A)R in the villous cytotrophoblastic and syncytiotrophoblastic cells was further confirmed in primary culture. A significantly (P < 0.05) decreased expression of SERT mRNA (56.3%) and protein (79.7%), and 5-HT(2A)R mRNA (79.1%) and protein (29.1%) was observed in placental tissues from GDM compared with non-GDM pregnancies. These data suggest that SERT and 5-HT(2A)R might be implicated in the aetiology of GDM. Moreover, the presence of SERT and 5-HT(2A)R in villous trophoblastic cells argues in favour of an important role of serotonin in human placental function.

Brain serotonin transporter in human methamphetamine users

Research on methamphetamine (MA) toxicity primarily focuses on the possibility that some of the behavioural problems in human MA users might be caused by damage to brain dopamine neurones. However, animal data also indicate that MA can damage brain serotonin neurones, and it has been suggested that cognitive problems and aggression in MA users might be explained by serotonergic damage. As information on the brain serotonin system in human MA users is fragmentary, our objective was to determine whether protein levels of serotonin transporter (SERT), a key marker of serotonin neurones, are decreased in brain of chronic MA users. SERT immunoreactivity was measured using an immunoblotting procedure in autopsied brain of 16 chronic MA users testing positive for the drug in blood and brain and matched controls. SERT levels were non-significantly decreased (-14% to -33%) in caudate, putamen and thalamus (normal in hippocampus), and, unlike the robust striatal dopamine reduction, there was marked overlap between control and MA user ranges. Concentrations of SERT were significantly decreased (-23% to -39%) in orbitofrontal and occipital cortices (normal in frontopolar and temporal cortices). Our data suggest that MA might modestly damage brain serotonin neurones and/or inhibit SERT protein expression, with cerebral cortex being more affected than sub-cortical regions. The SERT reduction in orbitofrontal cortex complements other data suggesting involvement of this area in MA-related behaviour. Decreased brain SERT could also be related to the clinical finding that treatment with a selective serotonin re-uptake inhibitor might increase relapse to MA.

Antidepressants modulate the in vitro inhibitory effects of propofol and ketamine on norepinephrine and serotonin transporter function

Norepinephrine transporter (NET) and serotonin transporter (SERT) proteins regulate norepinephrine (NE) and serotonin via their reuptake function and are targets of antidepressants action. Several intravenous (IV) anesthetics have been shown to inhibit NET and SERT. The interactions between antidepressants and anesthetics on transporter function, however, are not well studied. We examined the effect of different IV anesthetics on NET and SERT function, with and without chronic antidepressant pretreatment, by measuring NE or 5-hydroxytryptamine (5-HT) uptake and determined NET and SERT protein expression via immunoblotting. Both ketamine and propofol inhibited NET dose-dependently (propofol 10 −4M −22% ± 5.6%, and propofol 10 −3M −35% ± 5.7%; ketamine 10 −4M −23% ± 4.1% and ketamine 10 −3M −73% ± 2.9%); and SERT (propofol 10 −4M −11% ± 4.3% and propofol 10 −3M −23% ± 3.8%; ketamine 10 −4M −29% ± 5.2% and ketamine 10 −3M −63% ± 6.4%). Etomidate and thiopental had no effect on either NET or SERT function. Desipramine and fluoxetine, specific inhibitors of NET and SERT, respectively, both enhanced the inhibitory effects of propofol but reduced the inhibitory effects of ketamine on NET and SERT functions. IV anesthetics treatment did not change transporter protein expression in the presence of its respective inhibitor. Our results demonstrate that both ketamine and propofol inhibited SERT and NET function, but the inhibition was differentially modulated by antidepressants. Therefore, in the clinical context, this would suggest that patients receiving antidepressant treatments might have altered response to IV anesthetics in an agent-specific manner.

Expression of serotonin transporters by peripheral blood mononuclear cells of rhesus monkeys ( Macaca mulatta)

It has been well established that serotonin (5-hydroxytryptamine, 5-HT) plays a key role in neuro-endocrine-immune networks, mostly through its receptors and/or transporters. Although the presence of 5-HT receptor mRNAs in peripheral blood mononuclear cells (PBMCs) of rhesus monkeys has been reported, there is little information about serotonin transporter (SERT) expression by these cells. To examine SERT expression at the transcription and translation level, one-step RT-PCR, confocal microscopy and flow cytometry were used to detect SERT mRNA and protein expression by rhesus monkey PBMCs. It was found that SERT mRNA could be detected by RT-PCR from all of the rhesus macaque PBMC RNA samples and the nucleotide sequence of the amplicons was identical to the published SERT mRNA sequence. Low level SERT immunoreactivity was also demonstrated on the surface of rhesus PBMCs by confocal microscopy. Almost all lymphocytes and most monocytes were positive for SERT by flow cytometry. In the 2 rhesus macaques examined by multicolor flow cytometry, SERT bright cells were more than 84%, 94%, and 96% among CD20+, CD3+, and CD3+CD4+ lymphocytes respectively. These data demonstrate expression of SERT by rhesus macaque PBMCs, and indicate that rhesus macaques would be suitable models to test the in vivo immune regulatory effects of 5-HT or drugs targeting SERT.

Serotonin transporter protein in pulmonary hypertensive rats treated with atorvastatin

HMG-CoA-reductase inhibitors (statins) influence lipid metabolism and have pleiotropic effects. Several statins reduce various forms of pulmonary hypertension (PH) in animal models. The relationship between atorvastatin and expression of serotonin transporter protein (5-HTT) remains unknown. This study focused on the effects of atorvastatin on the course of monocrotaline (MCT)-induced PH and its relation to 5-HTT expression. Male Sprague-Dawley rats were challenged with MCT with or without subsequent daily oral treatment with 0.1, 1, and 10 mg/kg of atorvastatin for 28 days. Over the 4-wk course, the progression of PH was followed by transthoracic echocardiography [pulmonary artery pressure was assessed by pulmonary artery flow acceleration time (PAAT), an estimate reciprocal to pulmonary artery pressure], and, at the end of the 4-wk course, invasive right ventricular pressure, right ventricular weight, quantitative morphology, and 5-HTT expression were measured. MCT caused significant PH as early as 7 days after injection. Atorvastatin treatment increased PAAT and reduced right ventricular pressure, right ventricular hypertrophy, and vascular remodeling over the 4-wk course. MCT challenge was associated with increased pulmonary vascular 5-HTT expression, and atorvastatin treatment reduced the 5-HTT expression. MCT-induced PH over the course of 4 wk can be easily followed by transthoracic echocardiography, and atorvastatin is effective in reducing the PH. Atorvastatin's effects are associated with a decrease of 5-HTT expression.

Phosphorylation of Threonine Residue 276 Is Required for Acute Regulation of Serotonin Transporter by Cyclic GMP

Cellular protein kinases, phosphatases, and other serotonin transporter (SERT) interacting proteins participate in several signaling mechanisms regulating SERT activity. The molecular mechanisms of protein kinase G (PKG)-mediated SERT regulation and the site of transporter phosphorylation were investigated. Treatment of rat midbrain synaptosomes with 8-bromo-cGMP increased SERT activity, and the increase was selectively blocked by PKG inhibitors. The V(max) value for serotonin (5-HT) transport increased following cGMP treatment. However, surface biotinylation studies showed no change in SERT surface abundance following PKG activation. (32)P metabolic labeling experiments showed increased SERT phosphorylation in the presence of cGMP that was abolished by selectively inhibiting PKG. Phosphoamino acid analysis revealed that cGMP-stimulated native SERT phosphorylation occurred only on threonine residues. When added to CHO-1 cells expressing SERT, 8-bromo-cGMP stimulated 5-HT transport and SERT phosphorylation. Mutation of SERT threonine 276 to alanine completely abolished cGMP-mediated stimulation of 5-HT transport and SERT phosphorylation. Although the T276A mutation had no significant effect on 5-HT transport or SERT protein expression, mutation to aspartate (T276D) increased the level of 5-HT uptake to that of cGMP-stimulated 5-HT uptake in wild-type SERT-expressing cells and was no longer sensitive to cGMP. These findings provide the first identification of a phosphorylation site in SERT and demonstrate that phosphorylation of Thr-276 is required for cGMP-mediated SERT regulation. They also constitute the first evidence that in the central nervous system PKG activation stimulates endogenous SERT activity by a trafficking-independent mechanism.

Serotonin transporter gene polymorphisms in irritable bowel syndrome

Irritable bowel syndrome (IBS) is a symptomatic motility and sensory disorder of the lower gastrointestinal (GI) tract and is characterized by abdominal pain/ discomfort, bloating and altered bowel activity. It is usually classified according to the predominant bowel habit – IBS with constipation (C-IBS), IBS with diarrhoea (D-IBS) or IBS with alternating symptoms of both constipation and diarrhoea. Despite intensive research, the complicated and multifactorial pathophysiology underlying IBS is not completely understood. Recently, there is growing emphasis on identifying specific biological pathways that contribute to the complex pathophysiology of this disease.

Serotonin transporter expression is not sufficient to confer cytotoxicity to 3,4-methylenedioxymethamphetamine (MDMA) in vitro

There is much evidence from animal studies that the recreational drug MDMA is a selective toxin which damages serotonin nerve terminals and axons. These in vivo studies show that an interaction between MDMA and the serotonin transporter protein (SERT) is the .rst step in toxicity. To further our understanding of the biochemical processes of MDMA toxicity we wished to use an in vitro model for toxicity. We produced two COS-7 cell lines with different levels of expression of recombinant rat SERT, as determined by 5-HT uptake assays, and compared them to human SERT expressing JAR cells and to untransfected COS-7 cells which do not express SERT. Cultured cells were exposed to MDMA (0.1 microM-1 mM) for 24 or 48 h at 37 degrees C before assessing cytotoxicity by LDH release and MTT turnover. Only at the highest concentration used, 1 mM, was MDMA cytotoxic, and this toxicity was found in all cell lines. Cytotoxicity caused by 48 h exposure to 1 mM MDMA at 37 degrees C was not related to the level of SERT expression, not blocked by the SERT-blocking drugs paroxetine or fluoxetine and not enhanced, in JAR cells, by forskolin preincubation that increased 5-HT uptake capacity by 50%. We conclude that SERT expression is not sufficient to confer MDMA toxicity to cell lines. Therefore SERT-expressing cell lines do not offer a simple model system to elucidate the mechanisms underlying MDMA toxicity.

Effects of continuous administration of paroxetine on ligand binding site and expression of serotonin transporter protein in mouse brain

Selective serotonin reuptake inhibitors (SSRIs), such as paroxetine, are utilized in the treatment of depression and anxiety disorders. Although SSRIs potently interfere with the activity of brain serotonin transporter (SERT) after acute treatment, clinical improvement of psychiatric diseases is observed only after the repeated administration for several weeks (2–6 weeks). The present study was undertaken to investigate the effects of continuous administration of paroxetine on specific [ 3H]paroxetine binding sites and expression of SERT protein in mouse brain. Mice continuously and subcutaneously received paroxetine at doses of 2.67 or 13.3 μmol/kg/day for 21 days by using osmotic minipumps, and the steady-state plasma drug levels were within the range of reported concentrations in the clinical therapy. Continuous administration of paroxetine at theses doses produced significant (25–46%) reduction of [ 3H]paroxetine binding in each brain region (cerebral cortex, striatum, hippocampus, thalamus, midbrain) of mice. In Western blot analysis, expression levels of SERT protein in the thalamus and midbrain of mice were significantly (51% and 61%, respectively) decreased on day 21 after the implantation of minipumps at the higher dose. In conclusion, this study has firstly shown that continuous administration of paroxetine induces significant reduction of not only ligand binding sites of SERT but the protein expression level in mouse brain. Such down-regulation of SERT may partly underlie the therapeutic effect of long-term treatment with SSRIs in human.

Serotonergic‐like progenitor cells propagated from neural stem cells in vitro: survival with SERT protein expression following implantation into brains of mice lacking SERT

Neural stem cells (NSCs) obtained from the midbrain region of embryonic (E14) mice were initially cultured with basic fibroblast growth factor (bFGF), Sonic hedgehog, and FGF-8 in a serum-free N-2 culture medium to foster differentiation into a serotonergic-like phenotype. During the initial differentiating phase, these progenitor cells expressed En1, Pax3, and Pax5 mRNA. Subsequently, a single serotonin [5-hydroxytryptamine (5-HT)] and tryptophan hydroxylase-positive clone was isolated, which gave rise to cells that developed serotonergic properties. Sixty percent of these progenitor cells expressed the serotonin transporter (SERT), as indicated by specific ligand binding of [125I]-RTI-55. To further evaluate SERT functionality, we showed that these progenitor cells possessed specific [3H]-5-HT uptake activity. Implantation of the serotonergic-like progenitors into the hippocampus of adult mice genetically lacking SERT was followed by migration of these cells into adjacent brain regions, and survival of the cells at 8 weeks was accompanied by a gradual increase in density of SERT protein expression, which was not found in vehicle-injected, control mice. These findings suggest that this serotonergic-like NSC model will be a useful contribution to the development of cell biotechnology in regard to the expression of missing genes such as SERT in the adult brain.

Peritoneal macrophages express the serotonin transporter

Although it is known that macrophages take up serotonin, a specific monoamine transporter has not been identified in macrophages. In this study, mRNA coding for the serotonin transporter (SERT) was detected with the reverse transcription–polymerase chain reaction (RT-PCR) in recruited mouse peritoneal macrophages. Sequencing confirmed the identity of the RT-PCR product to mouse SERT mRNA. SERT protein was detected by Western blotting. Macrophage activation with lipopolysaccharide had no effect on expression of SERT mRNA or protein. Consistent with expression of a functional SERT, specific uptake of 3H-serotonin in macrophages was sodium dependent and inhibited by fluoxetine (IC 50 6.9 nM) and desipramine (IC 50 32 nM) but not by nisoxetine or reserpine.

Substituted Amphetamines That Produce Long‐Term Serotonin Depletion in Rat Brain (“Neurotoxicity”) Do Not Decrease Serotonin Transporter Protein Expression

Administration of high-dose D-fenfluramine (D-FEN) or parachloroamphetamine (PCA) produces long-lasting decreases in serotonin transporter (SERT) binding and tissue levels of serotonin (5-HT) in rat forebrain. These changes have been viewed as evidence for 5-HT neurotoxicity, but few studies have measured SERT protein levels. Thus, in the present study we determined the effect of high-dose D-FEN or PCA, administered according to a "neurotoxic" dosing regimen, on the density of SERT sites using ligand binding methods and on SERT protein levels using Western blots. Rats were sacrificed 2 days and 2 weeks after administration of drug or saline. The density of SERT was determined in homogenates of caudate and whole brain minus caudate. d-FEN and PCA decreased SERT binding by 30 to 60% in both tissues and at both time points. Similarly, D-FEN and PCA administration profoundly decreased tissue 5-HT and 5-HIAA in frontal cortex. Despite the large decreases in SERT binding and depletion of tissue 5-HT that occurred with d-FEN administration, SERT protein expression, as determined by Western blot analysis, did not change in either tissue or time point. PCA administration decreased SERT protein by about 20% only at the 2-day point in the caudate. Drug treatments did not change expression of glial fibrillary acidic protein (GFAP), a hallmark indicator of neuronal damage, in whole brain minus caudate in the 2-week group. These results support the hypothesis that D-FEN- and PCA-induced decreases in tissue 5-HT and SERT binding sites reflect neuroadaptive changes rather than neurotoxic effects.

High‐dose fenfluramine administration decreases serotonin transporter binding, but not serotonin transporter protein levels, in rat forebrain

Administration of D-fenfluramine (D-FEN) or parachloroamphetamine (PCA) can produce long-lasting decreases in serotonin transporter (SERT) binding and tissue levels of serotonin (5-HT) in rat forebrain. These changes have been viewed as evidence for 5-HT neurotoxicity, but no studies have measured SERT protein levels. In the present study, we determined the effect of high-dose D-FEN or PCA, administered according to a "neurotoxic" dosing regimen, on the density of SERT sites using ligand binding methods and on SERT protein levels using Western blots. Rats were sacrificed 2 days and 2 weeks after administration of drug or saline. The density of SERT was determined in homogenates of caudate and whole brain minus caudate. D-FEN and PCA decreased SERT binding by 30-60% in both tissues and at both time points. Similarly, D-FEN and PCA administration profoundly decreased tissue 5-HT and 5-HIAA in frontal cortex. Despite the large decreases in SERT binding and depletion of tissue 5-HT that occurred with D-FEN administration, SERT protein expression, as determined by Western blot analysis, did not change in either tissue or time point. PCA administration decreased SERT protein by about 20% only at the 2-day point in the caudate. Drug treatments did not change expression of glial fibrillary acidic protein (GFAP), a hallmark indicator of neuronal damage, in whole brain minus caudate in the 2-week group. These results support the hypothesis that decreases in tissue 5-HT and SERT binding sites induced by D-FEN and PCA reflect neuroadaptive changes, rather than neurotoxic effects.

Localization of norepinephrine and serotonin transporter in mouse brain capillary endothelial cells.

Monoamines function as a vasoactive modulator in the central nervous system (CNS) and are believed to regulate blood-brain barrier (BBB) function. Although monoamine transport is an essential process for regulating the extracellular monoamine concentration, the transport systems for monoamines at the BBB are poorly understood. mRNA expression of norepinephrine transporter (NET) and serotonin transporter (SERT) has been detected in a conditionally immortalized mouse brain capillary endothelial cell line (TM-BBB4) used as an in vitro model of the BBB, whereas no dopamine transporter (DAT) was detected. Western blot analysis showed the expression of NET and SERT protein in the membrane fraction of mouse brain capillaries and TM-BBB4 cells. Immunohistochemical analysis revealed that NET and SERT are localized at the brain capillaries in the mouse cerebral cortex, and suggests that NET is localized at the abluminal side of brain capillary endothelial cells, and SERT is localized at the luminal and abluminal sides. NET and SERT expressed at the BBB may be involved in the inactivation of monoamines released from neurons around the BBB.

Expression of serotonin transporter protein in developing rat brain

Serotonin transporter (5-HTT), a transmembrane protein, has been shown in adult brain to be distributed not only on synaptic terminals but to a great extent on axons as well. Here we report the ontogeny of 5-HTT and its relationship with serotonin (5-HT) neurons using established 5-HTT and 5-HT antibodies. Both 5-HTT- and 5-HT-immunostaining (-im) appear in 5-HT neurons at embryonic day 12 (E12) in rostral raphe nuclei (RRN). Soon after appearing, 5-HTT-im is highly expressed on axons, similar to adult expression. However, in contrast to adult, 5-HTT-im also outlines the soma-dendrites. Rich 5-HTT-im appears along the entire length of projecting axons, extending to the growth tip. In the next 2 days, intensive 5-HTT-im axons from RRN travel a course preferentially in the floor plate and later, the medial forebrain bundle trajectory. A group of new 5-HT-im neurons and 5-HTT-im axons appear at E13 in caudal raphe nuclei. At E16–18, taking the exact trajectory course of 5-HT axons, 5-HTT-im axons reach ganglionic eminence, olfactory bulb, and cortex and disperse into many brain regions in E18-20. No 5-HTT-im cell bodies were seen in nigral, locus ceruleus, or hypothalamus. However, the transient expression of 5-HTT on non-serotonergic system was seen in cortical and striatal neuroepithelia at E12 and sensory thalamic pathways at P0–P10. Prominent 5-HTT-im fibers in thalamocortical bundles project from sensory thalamic nuclei through reticular nucleus, internal capsule bundle and form barrels in somatosensory cortices. No 5-HTT-im was seen in glia-like cells using currently available antibody. These observations indicate that 5-HTT is: (a) associated preferentially with 5-HT neurons in brainstem, (b) temporally co-expressed with 5-HT in 5-HT neurons, (c) expressed on axons prior to synaptical sites at target neurons, which strongly indicates a volumic (extrasynaptic) transmission, (d) expressed in non-5-HT neurons within a specific window, which may affect the development of the systems “borrowing” the 5-HT. The early appearance of 5-HTT may also endow functionality as well as vulnerabilities of 5-HT, sensory thalamic, and cortical neurons to 5-HTT targeting drugs during pregnancy and after birth.

Antiphosphatidylserine antibody induces intrauterine growth retardation in BALB/C mice

Recent studies reveal that serotonin (5-hydroxytryptamine, 5-HT) might play a role in the aetiology of gestational diabetes mellitus (GDM). The involvement of the serotonin transporter (SERT) and 5-HT2A receptor (5-HT2AR) in diabetes has also been suggested. However, placental SERT and 5-HT2AR have never been studied in GDM-complicated pregnancies. The aim of this study was to investigate the effect of GDM on the expression of both placental SERT and 5-HT2AR. First, immunohistochemical analysis demonstrated the presence of SERT and 5-HT2AR proteins in the villous trophoblast and the fetal capillary endothelium of normal term placental tissue. Protein and mRNA expression of SERT and 5-HT2AR in the villous cytotrophoblastic and syncytiotrophoblastic cells was further confirmed in primary culture. A significantly (P < 0.05) decreased expression of SERT mRNA (56.3%) and protein (79.7%), and 5-HT2AR mRNA (79.1%) and protein (29.1%) was observed in placental tissues from GDM compared with non-GDM pregnancies. These data suggest that SERT and 5-HT2AR might be implicated in the aetiology of GDM. Moreover, the presence of SERT and 5-HT2AR in villous trophoblastic cells argues in favour of an important role of serotonin in human placental function.