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      Identification of 5-Hydroxytryptamine-Producing Cells by Detection of Fluorescence in Paraffin-Embedded Tissue Sections

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          Abstract

          5-Hydroxytryptamine (5-HT) produced by enterochromaffin (EC) cells is an important enteric mucosal signaling ligand and has been implicated in several gastrointestinal diseases, including inflammatory bowel disease and functional disorders such as irritable bowel syndrome. The present study reports a new, simple and rapid visualization method of 5-HT-producing EC cells utilizing detection of fluorescence in paraffin-embedded tissue sections after formalin fixation. In human samples, there was a high incidence of fluorescence + cells in the 5-HT + cells in the pyloric, small intestinal and colonic glands, while co-localization was lacking between fluorescence + and gastrin + cells in the pyloric and small intestinal glands. Fluorescence + EC cells were detected in the colon of mice and rats. Fluorescence + cells were also observed in 5-HT + β cells in the pancreatic islets of Langerhans in pregnant mice, while non-pregnant mouse pancreatic islet cells showed no 5-HT immunoreactivity or fluorescence. These results suggest that fluorescence + cells are identical to 5-HT + cells, and the source of fluorescence may be 5-HT itself or molecules related to its synthesis or degradation. This fluorescence signal detection method may be applicable for monitoring of inflammatory status of inflammatory bowel diseases in both the experimental and clinical settings.

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          Synthesis of serotonin by a second tryptophan hydroxylase isoform.

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            Serotonin Regulates Pancreatic β-Cell Mass during Pregnancy

            During pregnancy, the energy requirements of the fetus impose changes in maternal metabolism. Increasing insulin resistance in the mother maintains nutrient flow to the growing fetus, while prolactin and placental lactogen counterbalance this resistance and prevent maternal hyperglycemia by driving expansion of the maternal population of insulin-producing β-cells1–3. However, the exact mechanisms by which the lactogenic hormones drive β-cell expansion remain uncertain. Here we show that serotonin acts downstream of lactogen signaling to drive β-cell proliferation. Serotonin synthetic enzyme Tph1 and serotonin production increased sharply in β-cells during pregnancy or after treatment with lactogens in vitro. Inhibition of serotonin synthesis by dietary tryptophan restriction or Tph inhibition blocked β-cell expansion and induced glucose intolerance in pregnant mice without affecting insulin sensitivity. Expression of the Gαq-linked serotonin receptor Htr2b in maternal islets increased during pregnancy and normalized just prior to parturition, while expression of the Gαi-linked receptor Htr1d increased at the end of pregnancy and postpartum. Blocking Htr2b signaling in pregnant mice also blocked β-cell expansion and caused glucose intolerance. These studies reveal an integrated signaling pathway linking β-cell mass to anticipated insulin need during pregnancy. Modulators of this pathway, including medications and diet, may affect the risk of gestational diabetes4.
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              A unique central tryptophan hydroxylase isoform

              Biochemical Pharmacology, 66(9), 1673-1680
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                Author and article information

                Journal
                Eur J Histochem
                Eur J Histochem
                EJH
                European Journal of Histochemistry : EJH
                PAGEPress Publications, Pavia, Italy
                1121-760X
                2038-8306
                29 September 2016
                10 August 2016
                : 60
                : 3
                : 2684
                Affiliations
                [1 ]R&D Group, Olympus Corporation , Tokyo
                [2 ]Laboratory of Veterinary Pathology, Tokyo University of Agriculture and Technology , Tokyo
                [3 ]United Graduate School of Veterinary Sciences, Gifu University , Japan
                Author notes
                Laboratory of Veterinary Pathology, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu-shi, Tokyo 183-8509, Japan. +81-42-367-5771. mshibuta@ 123456cc.tuat.ac.jp

                Contributions: YK, MS, contribution on the conception and design (detection of fluorescence), development of methodology (detection of fluorescence), acquisition of data (fluorescence), analysis and interpretation of the data (fluorescence and immunohistochemistry), manuscript drafting, and administrative, technical, or material support; NO, YW, contributed on the acquisition of data (immunohistochemistry), analysis and interpretation of the data (immunohistochemistry), manuscript review, technical supports; MS, supervision of the study. All authors have read and approved the final manuscript.

                Disclosures: YK, NO, are employees of Olympus Corporation, whose product lines include microscope and endoscope systems. The views and opinions expressed in this article are those of the authors and do not necessarily reflect those of Olympus Corporation. YW, MS declare no conflict of interest.

                Article
                10.4081/ejh.2016.2684
                5062634
                27734992
                2c789a69-710f-4178-9a66-d8b6ac51a035
                ©Copyright Y. Kaneko et al.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 15 May 2016
                : 18 August 2016
                Page count
                Figures: 9, Tables: 0, Equations: 0, References: 31, Pages: 6
                Categories
                Original Paper

                Clinical chemistry
                5-hydroxytryptamine,enterochromaffin cells,fluorescence,paraffin-embedded tissue sections,formalin fixation

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