title: Vesicular inhibitory amino acid transporter is present in glucagon-containing secretory granules in alphaTC6 cells, mouse clonal alpha-cells, and alpha-cells of islets of Langerhans.
abstract: Islets of Langerhans contain gamma-aminobutyrate (GABA) and may use it as an intercellular transmitter. In beta-cells, GABA is stored in synaptic-like microvesicles and secreted through Ca(2+)-dependent exocytosis. Vesicular inhibitory amino acid transporter (VIAAT), which is responsible for the storage of GABA and glycine in neuronal synaptic vesicles, is believed to be responsible for the storage and secretion of GABA in beta-cells. However, a recent study by Chessler et al. indicated that VIAAT is expressed in the mantle region of islets. In the present study, we investigated the precise localization of VIAAT in rat islets of Langerhans and clonal islet cells and found that it is present in alpha-cells, a minor population of F-cells and alphaTC6 cells, and clonal alpha-cells but not in beta-cells, delta-cells, or MIN6 m9-cells (clonal beta-cells). Combined biochemical, immunohistochemical, and electronmicroscopical evidence indicated that VIAAT is specifically localized with glucagon-containing secretory granules in alpha-cells. ATP-dependent uptake of radiolabeled GABA, which is energetically coupled with a vacuolar proton pump, was detected in digitonin-permeabilized alphaTC6 cells as well as in MIN6 m9 cells. These results demonstrate that functional neuronal VIAAT is present in glucagon-containing secretory granules in alpha-cells and suggest that the ATP-dependent GABA transporter in beta-cells is at least immunologically distinct from VIAAT. Because glucagon-containing secretory granules also contain vesicular glutamate transporter and store L-glutamate, as demonstrated by Hayashi et al., the present results suggest more complex features of the GABAergic phenotype of islets than previously supposed.
authors.affiliation: Department of Biochemistry, Faculty of Pharmaceutical Sciences, Okayama University, Okayama, Japan.
authors.forename: Mitsuko
authors.initials: M
authors.lastname: Hayashi
authors.forename: Masato
authors.initials: M
authors.lastname: Otsuka
authors.forename: Riyo
authors.initials: R
authors.lastname: Morimoto
authors.forename: Akiko
authors.initials: A
authors.lastname: Muroyama
authors.forename: Shunsuke
authors.initials: S
authors.lastname: Uehara
authors.forename: Akitsugu
authors.initials: A
authors.lastname: Yamamoto
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Moriyama
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D026905:Amino Acid Transport Systems; D000818:Animals; D019556:COS Cells; D002352:Carrier Proteins / Q000032:analysis* / Q000378:metabolism; D002999:Clone Cells; D005455:Fluorescent Antibody Technique; D005934:Glucagon / Q000032:analysis*; D007515:Islets of Langerhans / Q000737:chemistry* / Q000166:cytology / Q000378:metabolism; D008297:Male; D026901:Membrane Transport Proteins; D051379:Mice; D016716:PC12 Cells; D051381:Rats; D017208:Rats, Wistar; D022142:Secretory Vesicles / Q000737:chemistry* / Q000378:metabolism; D050599:Vesicular Glutamate Transport Protein 2; D050495:Vesicular Inhibitory Amino Acid Transport Proteins; D033921:Vesicular Transport Proteins; D005680:gamma-Aminobutyric Acid / Q000493:pharmacokinetics
chemical_list: D026905:Amino Acid Transport Systems; D002352:Carrier Proteins; D026901:Membrane Transport Proteins; C493477:Slc17a6 protein, mouse; C493475:Slc17a6 protein, rat; C118623:Slc32a1 protein, rat; D050599:Vesicular Glutamate Transport Protein 2; D050495:Vesicular Inhibitory Amino Acid Transport Proteins; D033921:Vesicular Transport Proteins; C492244:Viaat protein, mouse; D005680:gamma-Aminobutyric Acid; D000255:Adenosine Triphosphate; D005934:Glucagon