Vesicular storage and secretion of L-glutamate from glucagon-like peptide 1-secreting clonal intestinal L cells

Shunsuke Uehara, Sun Kyung Jung, Riyo Morimoto, Shigeo Arioka, Takaaki Miyaji, Narinobu Juge, Miki Hiasa, Kahori Shimizu, Akinori Ishimura, Masato Otsuka, Akitsugu Yamamoto, Pierre Maechler, Yoshinori Moriyama

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Vesicular glutamate transporter (VGLUT) is responsible for the vesicular storage of L-glutamate, and plays an essential role in glutamate-mediated intercellular signal transmission in the CNS and in some neuroendocrine cells. Intestinal L cells are the glucose-responsive neuroendocrine cells responsible for the secretion of glucagon-like peptide 1 (GLP-1). We have shown that intestinal L cells express VGLUT2, a VGLUT isoform, which suggests that L cells secrete L-glutamate. In the present study, we investigated this possibility using GLUTag mouse clonal L cells. RT-PCR and northern blot analyses revealed expression of the VGLUT1 and VGLUT2 genes, but not of the VGLUT3 gene. Western blot analysis revealed immunological counterparts for VGLUT2, whereas an immunological counterpart of VGLUT1 was not detected. Indirect immunofluorescence microscopy revealed a punctate distribution of VGLUT2 immunoreactivity throughout the cells, which co-localized with GLP-1. Double-labeling immunoelectronmicroscopy confirmed the association of VGLUT2 with GLP-1-containing secretory granules. The membrane fraction exhibited ATP-dependent L-glutamate uptake, which was sensitive to bafilomycin A1 (a vacuolar proton ATPase inhibitor) and Evans blue (a VGLUT inhibitor) but insensitive to D,L-aspartate. Upon depolarization with KCl, GLUTag cells secreted appreciable amounts of L-glutamate and GLP-1. D-Glucose and methyl-α-D-glucopyranoside, stimulators of exocytosis of GLP-1, also triggered the secretion of L-glutamate. The L-glutamate secretion was partially dependent on Ca2+ and sensitive to bafilomycin A1. These results demonstrated that GLUTag cells stored L-glutamate in secretory granules and secreted it with GLP-1 by exocytosis. As GLUTag cells and intestinal L cells express kainate receptors and plasma membrane glutamate transporters, these results support the concept of L-glutamate-mediated intercellular signaling in the vicinity of intestinal L cells.

Original languageEnglish
Pages (from-to)550-560
Number of pages11
JournalJournal of Neurochemistry
Volume96
Issue number2
DOIs
Publication statusPublished - Jan 2006

Fingerprint

Enteroendocrine Cells
Glucagon-Like Peptide 1
Glutamic Acid
Vesicular Glutamate Transport Proteins
Neuroendocrine Cells
Exocytosis
Secretory Vesicles
Genes
Vacuolar Proton-Translocating ATPases
Kainic Acid Receptors
Amino Acid Transport System X-AG
D-Aspartic Acid
Glucose
Evans Blue
Membrane Transport Proteins
Depolarization
Cell membranes
Indirect Fluorescent Antibody Technique
Fluorescence Microscopy
Aspartic Acid

Keywords

  • Blood glucose
  • Glucagon-like peptide 1
  • GLUTag
  • L cell
  • L-glutamate
  • Vesicular glutamate transporter

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Vesicular storage and secretion of L-glutamate from glucagon-like peptide 1-secreting clonal intestinal L cells. / Uehara, Shunsuke; Jung, Sun Kyung; Morimoto, Riyo; Arioka, Shigeo; Miyaji, Takaaki; Juge, Narinobu; Hiasa, Miki; Shimizu, Kahori; Ishimura, Akinori; Otsuka, Masato; Yamamoto, Akitsugu; Maechler, Pierre; Moriyama, Yoshinori.

In: Journal of Neurochemistry, Vol. 96, No. 2, 01.2006, p. 550-560.

Research output: Contribution to journalArticle

Uehara, S, Jung, SK, Morimoto, R, Arioka, S, Miyaji, T, Juge, N, Hiasa, M, Shimizu, K, Ishimura, A, Otsuka, M, Yamamoto, A, Maechler, P & Moriyama, Y 2006, 'Vesicular storage and secretion of L-glutamate from glucagon-like peptide 1-secreting clonal intestinal L cells', Journal of Neurochemistry, vol. 96, no. 2, pp. 550-560. https://doi.org/10.1111/j.1471-4159.2005.03575.x
Uehara, Shunsuke ; Jung, Sun Kyung ; Morimoto, Riyo ; Arioka, Shigeo ; Miyaji, Takaaki ; Juge, Narinobu ; Hiasa, Miki ; Shimizu, Kahori ; Ishimura, Akinori ; Otsuka, Masato ; Yamamoto, Akitsugu ; Maechler, Pierre ; Moriyama, Yoshinori. / Vesicular storage and secretion of L-glutamate from glucagon-like peptide 1-secreting clonal intestinal L cells. In: Journal of Neurochemistry. 2006 ; Vol. 96, No. 2. pp. 550-560.
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