The afferent innervation of the gastrointestinal (GI) tract consists of intrinsic and extrinsic sensory neurons that respond to nutrients, chemicals or mechanical stimuli within the gut lumen. Most stimuli do not interact directly with the afferent nerves but instead activate specialised cells in the epithelium in a process of sensory transduction. It is thought that one of the first steps in this process is the release of serotonin (5-HT) from the enterochromaffin (EC) cells. The EC cells are a sub-type of enteroendocrine (EE) cells which are found among the enterocytes of the intestinal epithelium. The EC cells are responsible for the production and storage of the largest pool of 5 HT in the body. Released 5-HT can act on the intrinsic nerves and vagal endings. This review will focus on the role of 5-HT in sensory transduction and examine how the EC cell produces and releases 5-HT. We will explore recent developments that have helped to elucidate some of the proteins that allow EC cells to sense the luminal environment. Finally, we will highlight some of the findings from new studies using electrochemical techniques which allow the real-time recording of 5-HT concentrations near to the EC cell.

Diet-induced obesity is associated with changes in gastrointestinal function and induction of a mild inflammatory state. Serotonin (5-HT) containing enterochromaffin (EC) cells within the intestine respond to nutrients and are altered by inflammation. Thus, our aim was to characterize the uptake and release of 5-HT from EC cells of the rat ileum in a physiologically relevant model of diet-induced obesity. In chow-fed (CF) and Western diet-fed (WD) rats electrochemical methods were used to measure compression evoked (peak) and steady state (SS) 5-HT levels with fluoxetine used to block the serotonin reuptake transporter (SERT). The levels of mRNA for tryptophan hydroxylase 1 (TPH1) and SERT were determined by quantitative PCR, while EC cell numbers were determined immunohistochemically. In WD rats, the levels of 5-HT were significantly increased (SS: 19.2 ± 3.7 μm; peak: 73.5 ± 14.1 μm) compared with CF rats (SS: 12.3 ± 1.8 μm; peak: 32.2 ± 7.2 μm), while SERT-dependent uptake of 5-HT was reduced (peak WD: 108% of control versus peak CF: 212% control). In WD rats, there was a significant increase in TPH1 mRNA, a decrease in SERT mRNA and protein, and an increase in EC cells. In conclusion, our data show that foods typical of a Western diet are associated with an increased 5-HT availability in the rat ileum. Increased 5-HT availability is driven by the up-regulation of 5-HT synthesis genes, decreased re-uptake of 5-HT, and increased numbers and/or 5-HT content of EC cells which are likely to cause altered intestinal motility and sensation in vivo.

Serotonin (5-HT)-containing enterochromaffin (EC) cells of the intestine transduce chemical and mechanical stimuli from the intestinal lumen by releasing 5-HT on to afferent nerve terminals. Dysfunctional mucosal 5-HT signaling has been implicated in heightened visceral sensitivity and altered motility in patients with inflammatory bowel disease and in animal models. Our aim was to characterize the release and uptake of 5-HT in the mouse dextran sulfate sodium (DSS; 5% wt/vol) model of colitis. We made electrochemical recordings and used an ELISA assay to determine mucosal 5-HT release and uptake in untreated mice and mice with DSS-induced colitis. Peak and steady-state 5-HT concentrations were measured before and during blockade of the serotonin reuptake transporter (SERT) with 1 microM fluoxetine. Electrochemical recordings showed that colons from DSS-treated mice had roughly twice the steady-state levels of extracellular 5-HT and compression-evoked 5-HT release compared with untreated mice. Fluoxetine doubled the compression-evoked and steady-state 5-HT levels in control and DSS mice. These data were supported by ELISA assays, which showed enhanced 5-HT release during colitis, by immunohistochemical analyses, which showed increases in EC cell numbers, and by real-time PCR, which identified a decrease in SERT mRNA expression in the mucosa during colitis. These data are the first to demonstrate 5-HT release close to its release site and near its site of action during DSS-colitis. We conclude that DSS-colitis increases 5-HT availability primarily by an increase in the numbers of EC cells and/or of content of 5-HT in these EC cells.

The spatial localization of TRPC3 and associated channels, receptors, and calcium stores are integral for myoendothelial microdomain function. TRPC3 facilitates endothelial SK(Ca) and IK(Ca) activation, as key components of EDH-mediated vasodilator activity and for regulating mesenteric artery tone.

(5-HT) release and uptake measured by real-time electrochemical techniques in the rat ileum. Am J Physiol Gastrointest Liver Physiol 295: G1228 -G1236, 2008. First published October 16, 2008 doi:10.1152/ajpgi.90375.2008 is released from the enterochromaffin cells and plays an important role in regulating intestinal function. Although the release of 5-HT is well documented, the contribution of the serotonin reuptake transporter (SERT) to the levels and actions of 5-HT in the intestine is unclear. This study aimed to demonstrate real-time SERT activity in ileal mucosa and to assess the effects of SERT inhibition using fluoxetine. Electrochemical recordings were made from the mucosa in full-thickness preparations of rat ileum using a carbon fiber electrode to measure 5-HT oxidation current and a force transducer to record circular muscle (CM) tension. Compression of the mucosa stimulated a peak 5-HT release of 12 Ϯ 6 M, which decayed to 7 Ϯ 4 M. Blockade of SERT with fluoxetine (1 M) increased the peak compression-evoked release to 19 Ϯ 9 M, and the background levels of 5-HT increased to 11 Ϯ 7 M (P Ͻ 0.05, n ϭ 7). When 5-HT was exogenously applied to the mucosa, fluoxetine caused a significant increase in the time to 50% and 80% decay of the oxidation current. Fluoxetine also increased the spontaneous CM motility (P Ͻ 0.05; n ϭ 7) but did not increase the CM contraction-evoked 5-HT release (P Ͼ 0.05, n ϭ 5). In conclusion, this is the first characterization of the real-time uptake of 5-HT into the rat intestine. These data suggest that SERT plays an important role in the modulation of 5-HT concentrations that reach intestinal 5-HT receptors. electrochemistry; enterochromaffin cell; gastrointestinal tract; serotonin reuptake transporter is produced and stored in the enterochromaffin (EC) cells of the intestinal epithelium (19, 34). Released 5-HT acts as a sensory mediator and plays an important role in visceral sensation and in regulating gut function by activating 5-HT receptors on the afferent nerve terminals (1,22,26). Activation of the intrinsic sensory neurons of the enteric nervous system by 5-HT is responsible for initiating or enhancing local intestinal motor reflexes (11,(42)(43)(44) and for increasing secretory function (e.g., 39). EC-derived 5-HT also activates extrinsic afferent fibers and thus contributes to the relay of sensory information to the central nervous system (24,25,41).The actions of 5-HT are terminated by uptake via the serotonin reuptake transporter (SERT, a Na ϩ /Cl Ϫ -dependent transporter) (31). SERT is the target for many important therapeutic drugs such as Prozac (fluoxetine), a member of the serotonin-selective reuptake inhibitors. SERT has been localized to many intestinal epithelial cells using immunohistochemistry to detect protein (44) and Northern blots to detect mRNA (13). The SERT gene is subject to a number of naturally occurring polymorphisms in the coding and promoter regions, some of which have been linked to the symptoms of irritable bowel syndrome (IBS) (e.g., 46) or ...