Expression of the vesicular inhibitory amino acid transporter in pancreatic islet cells: distribution of the transporter within rat islets.
gamma-Aminobutyric acid (GABA) is stored in microvesicles in pancreatic islet cells. Because GAD65 and GAD67, which catalyze the formation of GABA, are cytoplasmic, the existence of an islet vesicular GABA transporter has been postulated. Here, we test the hypothesis that the putative transporter is the vesicular inhibitory amino acid transporter (VIAAT), a neuronal transmembrane transporter of GABA and glycine. We sequenced the human VIAAT gene and determined that the human and rat proteins share over 98% sequence identity. In vitro expression of VIAAT and immunoblotting of brain and islet lysates revealed two forms of the protein: an approximately 52-kDa and an approximately 57-kDa form. By immunoblotting and immunohistochemistry, we detected VIAAT in rat but not human islets. Immunohistochemical staining showed that in rat islets, the distribution of VIAAT expression parallels that of GAD67, with increased expression in the mantle. GABA, too, was found to be present in islet non-beta-cells. We conclude that VIAAT is expressed in rat islets and is more abundant in the mantle and that expression in human islets is very low or nil. The rat islet mantle differs from rat and human beta-cells in that it contains only GAD67 and relatively increased levels of VIAAT. Cells that express only GAD67 may require higher levels of VIAAT expression.
Chessler
Steven D
SD
Robert H. Williams Laboratory, Department of Medicine, University of Washington, Seattle, Washington 98195-7710, USA. chessler@u.washington.edu
Simonson
William T
WT
Sweet
Ian R
IR
Hammerle
Lisa P
LP
D000595:Amino Acid Sequence; D026905:Amino Acid Transport Systems; D000818:Animals; D001923:Brain Chemistry; D002352:Carrier Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D015870:Gene Expression; D005968:Glutamate Decarboxylase / Q000032:analysis; D005998:Glycine / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D007515:Islets of Langerhans / Q000737:chemistry*; D007527:Isoenzymes / Q000032:analysis; D051379:Mice; D008969:Molecular Sequence Data; D051381:Rats; D011913:Rats, Inbred BB; D016415:Sequence Alignment; D017421:Sequence Analysis; D014018:Tissue Distribution; D050495:Vesicular Inhibitory Amino Acid Transport Proteins; D033921:Vesicular Transport Proteins; D005680:gamma-Aminobutyric Acid / Q000032:analysis / Q000378:metabolism
D026905:Amino Acid Transport Systems; D002352:Carrier Proteins; D007527:Isoenzymes; C492245:SLC32A1 protein, human; C118623:Slc32a1 protein, rat; D050495:Vesicular Inhibitory Amino Acid Transport Proteins; D033921:Vesicular Transport Proteins; C492244:Viaat protein, mouse; D005680:gamma-Aminobutyric Acid; D005968:Glutamate Decarboxylase; C402565:glutamate decarboxylase 1; D005998:Glycine