Identification and characterization of a novel isoform of the vesicular gamma-aminobutyric acid transporter with glucose-regulated expression in rat islets.
Pancreatic islets are unique outside the nervous system in that they contain high levels of the inhibitory neurotransmitter gamma-aminobutyric acid (GABA), synthesized by the enzyme glutamic acid decarboxylase (GAD). Since the role that GABA plays in the islet and the mechanisms whereby the two major GAD isoforms (GAD65 and GAD67) function as diabetes-associated autoantigens are unknown, continued characterization of the islet GAD-GABA system is important. We previously demonstrated that the GABA and glycine transporter vesicular inhibitory amino acid transporter (VIAAT also known as VGAT) is present in rat islets. Here we identify a novel 52 kDa variant of VIAAT in rat islets: VIAAT-52 (V52). V52 is an amino-terminally truncated form of VIAAT (V57) that likely results from utilization of a downstream start site of translation. V57 and V52 display different patterns of post-translational modification and cellular expression. Our results have indicated that islet content of V52, but not V57, is responsive to changes in glucose concentration and other extracellular conditions. VIAAT is expressed in the islet alpha cells, but there have been conflicting findings regarding the presence of VIAAT in the beta cells. Here we have also provided additional evidence for the presence of VIAAT in islet beta cells and show that the beta cell line INS-1 expresses V57. V52 may be better adapted than V57 to the unique rat alpha cell GAD-GABA system, which lacks GAD65 and in which VIAAT traffics to secretory granules rather than just to synaptic microvesicles.
Suckow
A T
AT
Department of Medicine, University of California, San Diego, Leichtag Biomedical Research Building, 9500 Gilman Drive MC 0726, La Jolla, California 92093-0726, USA.
Sweet
I R
IR
Van Yserloo
B
B
Rutledge
E A
EA
Hall
T R
TR
Waldrop
M
M
Chessler
S D
SD
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D050485:GABA Plasma Membrane Transport Proteins / Q000737:chemistry* / Q000235:genetics*; D005786:Gene Expression Regulation / Q000187:drug effects*; D005947:Glucose / Q000494:pharmacology*; D007515:Islets of Langerhans / Q000187:drug effects* / Q000378:metabolism; D014176:Protein Biosynthesis; D020033:Protein Isoforms / Q000737:chemistry* / Q000235:genetics*; D051381:Rats; D011913:Rats, Inbred BB; D013347:Subcellular Fractions / Q000378:metabolism
D017931:DNA Primers; D050485:GABA Plasma Membrane Transport Proteins; D020033:Protein Isoforms; D005947:Glucose