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      Activation of 5-HT 4 receptors facilitates neurogenesis from transplanted neural stem cells in the anastomotic ileum

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          Abstract

          An orally administered serotonin-4 (5-HT 4) receptor agonist, mosapride citrate (MOS), promotes enteric neurogenesis in anastomoses after gut surgery. We performed gut surgery and transplanted 2 × 10 5 neural stem cells (NSCs) from the embryonic central nervous system after marking them with the cell linker, PKH26. We found that neurons differentiated from transplanted NSCs (PKH [+]) and newborn enteric neurons differentiated from mobilized (host) NSCs (YFP [+]) in the deep granulation tissue of the anastomotic ileum. MOS significantly increased the number of PKH (+) and YFP (+) neurons by 2.5-fold ( P < 0.005) ( n = 4). The distribution patterns of PKH (+) neurons and YFP (+) neurons were similar along the depth of the anastomosis. A 5-HT 4 receptor antagonist, SB-207266, abolished these effects of MOS ( n = 4). Our results indicate that neurogenesis from transplanted NSCs is potentiated by activation of 5-HT 4 receptors. Thus, a combination of drug administration and cell transplantation could be more beneficial than cell transplantation alone in treating Hirschsprung’s disease and related disorders.

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          Genetically Encoded Green Fluorescent Ca2+ Indicators with Improved Detectability for Neuronal Ca2+ Signals

          Masamichi Ohkura, Takuya Sasaki, Junko Sadakari … (2012)
          Imaging the activities of individual neurons with genetically encoded Ca2+ indicators (GECIs) is a promising method for understanding neuronal network functions. Here, we report GECIs with improved neuronal Ca2+ signal detectability, termed G-CaMP6 and G-CaMP8. Compared to a series of existing G-CaMPs, G-CaMP6 showed fairly high sensitivity and rapid kinetics, both of which are suitable properties for detecting subtle and fast neuronal activities. G-CaMP8 showed a greater signal (F max/F min = 38) than G-CaMP6 and demonstrated kinetics similar to those of G-CaMP6. Both GECIs could detect individual spikes from pyramidal neurons of cultured hippocampal slices or acute cortical slices with 100% detection rates, demonstrating their superior performance to existing GECIs. Because G-CaMP6 showed a higher sensitivity and brighter baseline fluorescence than G-CaMP8 in a cellular environment, we applied G-CaMP6 for Ca2+ imaging of dendritic spines, the putative postsynaptic sites. By expressing a G-CaMP6-actin fusion protein for the spines in hippocampal CA3 pyramidal neurons and electrically stimulating the granule cells of the dentate gyrus, which innervate CA3 pyramidal neurons, we found that sub-threshold stimulation triggered small Ca2+ responses in a limited number of spines with a low response rate in active spines, whereas supra-threshold stimulation triggered large fluorescence responses in virtually all of the spines with a 100% activity rate.
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            5-HT4 receptor-mediated neuroprotection and neurogenesis in the enteric nervous system of adult mice.

            Paul Gershon, Yung-Hui Kuan, Tso-Tsai Liu … (2009)
            Although the mature enteric nervous system (ENS) has been shown to retain stem cells, enteric neurogenesis has not previously been demonstrated in adults. The relative number of enteric neurons in wild-type (WT) mice and those lacking 5-HT(4) receptors [knock-out (KO)] was found to be similar at birth; however, the abundance of ENS neurons increased during the first 4 months after birth in WT but not KO littermates. Enteric neurons subsequently decreased in both WT and KO but at 12 months were significantly more numerous in WT. We tested the hypothesis that stimulation of the 5-HT(4) receptor promotes enteric neuron survival and/or neurogenesis. In vitro, 5-HT(4) agonists increased enteric neuronal development/survival, decreased apoptosis, and activated CREB (cAMP response element-binding protein). In vivo, in WT but not KO mice, 5-HT(4) agonists induced bromodeoxyuridine incorporation into cells that expressed markers of neurons (HuC/D, doublecortin), neural precursors (Sox10, nestin, Phox2b), or stem cells (Musashi-1). This is the first demonstration of adult enteric neurogenesis; our results suggest that 5-HT(4) receptors are required postnatally for ENS growth and maintenance.
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              Chemokines regulate the migration of neural progenitors to sites of neuroinflammation.

              Abdelhak Belmadani, Da B Tran, Dongjun Ren … (2006)
              Many studies have shown that transplanted or endogenous neural progenitor cells will migrate toward damaged areas of the brain. However, the mechanism underlying this effect is not clear. Here we report that, using hippocampal slice cultures, grafted neural progenitor cells (NPs) migrate toward areas of neuroinflammation and that chemokines are a major regulator of this process. Migration of NPs was observed after injecting an inflammatory stimulus into the area of the fimbria and transplanting enhanced green fluorescent protein (EGFP)-labeled NPs into the dentate gyrus of cultured hippocampal slices. Three to 7 d after transplantation, EGFP-NPs in control slices showed little tendency to migrate and had differentiated into neurons and glia. In contrast, in slices injected with inflammatory stimuli, EGFP-NPs migrated toward the site of the injection. NPs in these slices also survived less well. The inflammatory stimuli used were a combination of the cytokines tumor necrosis factor-alpha and interferon-gamma, the bacterial toxin lipopolysaccharide, the human immunodeficiency virus-1 coat protein glycoprotein 120, or a beta-amyloid-expressing adenovirus. We showed that these inflammatory stimuli increased the synthesis of numerous chemokines and cytokines by hippocampal slices. When EGFP-NPs from CC chemokine receptor CCR2 knock-out mice were transplanted into slices, they exhibited little migration toward sites of inflammation. Similarly, wild-type EGFP-NPs exhibited little migration toward inflammatory sites when transplanted into slices prepared from monocyte chemoattractant protein-1 (MCP-1) knock-out mice. These data indicate that factors secreted by sites of neuroinflammation are attractive to neural progenitors and suggest that chemokines such as MCP-1 play an important role in this process.
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                Author and article information

                Contributors
                Miyako Takaki: +81-744-29-8873 , mtakaki@naramed-u.ac.jp
                Journal
                Journal ID (nlm-ta): J Physiol Sci
                Journal ID (iso-abbrev): J Physiol Sci
                Title: The Journal of Physiological Sciences
                Publisher: Springer Japan (Tokyo )
                ISSN (Print): 1880-6546
                ISSN (Electronic): 1880-6562
                Publication date (Electronic): 3 September 2015
                Publication date PMC-release: 3 September 2015
                Publication date (Print): 2016
                Volume: 66
                Pages: 67-76
                Affiliations
                [ ]Department of Physiology II, Nara Medical University, School of Medicine, Kashihara, Nara Japan
                [ ]Department of Molecular Pathology, Nara Medical University, School of Medicine, Kashihara, Nara Japan
                [ ]Department of Orthopedic Surgery, Nara Medical University, School of Medicine, 840 Shijo-cho, Kashihara, Nara 634-8522 Japan
                [ ]Division of Homeostatic Development, Department of Developmental Physiology, National Institute for Physiological Sciences, Okazaki, Aichi Japan
                Article
                Publisher ID: 396
                DOI: 10.1007/s12576-015-0396-1
                PMC ID: 4676964
                PubMed ID: 26335766
                SO-VID: 4f2e9f3e-5aed-4223-976a-ea0e5ddc0650
                Copyright © © The Author(s) 2015
                License:

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

                History
                Date received : 26 July 2015
                Date accepted : 26 August 2015
                Funding
                Funded by: Grants-in-aid for Scientific Research (KAKENHI) from the Ministry of Education, Culture, Sports, Science, and Technology of Japan
                Award ID: 20659210, 23390330, 24650325, 26560280, 15H03057
                Award ID: 23591969
                Award Recipient :
                Categories
                Subject: Original Paper
                Custom metadata
                issue-copyright-statement © The Physiological Society of Japan and Springer Japan 2016

                ScienceOpen disciplines: Anatomy & Physiology
                Keywords: yfp,5-ht4 receptor,granulation tissue,myenteric neuron,neural stem cells
                Data availability:
                ScienceOpen disciplines: Anatomy & Physiology
                Keywords: yfp, 5-ht4 receptor, granulation tissue, myenteric neuron, neural stem cells

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