Abstract
GAD65 and GAD67 are two isoforms of the enzyme glutamic acid decarboxylase which catalyze the production of GABA from glutamate, primarily in the brain. However, GAD and GABA also prevail in the retina, testes and islets of Langerhans. The main function of GABA is in neurotransmission, and it is involved in paracrine signalling in islets, but has also been suggested to play a role as a trophic factor in synaptogenesis and to be an important metabolite feeding into the tricarboxylic acid cycle via the GABA-shunt. Both GAD isoforms are subject to regulation, e.g. by synaptic activity. GAD65 is regulated at the level of enzyme activity by association and dissociation from its cofactor, PLP, whereas GAD67 is controlled at the level of its mRNA. To study this process in further detail, we have isolated and characterized the 5'-flanking region of the rat GAD67 gene. We report the transcriptional initiation sites and promoter sequences important for expression in islet beta-cells and C6 glioma cells, and demonstrate that the GAD67 promoter harbors elements that are responsive to glucose in primary islet cells.
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