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Journal of Histochemistry and Cytochemistry, Vol. 48, 839-846, June 2000, Copyright © 2000, The Histochemical Society, Inc.

ARTICLE

Immunocytochemical Evidence Suggesting that Diamine Oxidase Catalyzes Biosynthesis of {gamma}-Aminobutyric Acid in Antropyloric Gastrin Cells

Jesper Hardta, Lars-Inge Larssonb, and David M. Hougaarda
a Department of Clinical Biochemistry, Statens Serum Institut, Copenhagen, Denmark
b Division of Cell Biology, Institute of Anatomy and Physiology, The Royal Veterinary and Agricultural University, Frederiksberg, Denmark

Correspondence to: David M. Hougaard, Dept. of Clinical Biochemistry, Statens Serum Institut, Artillerivej 5, DK-2300 Copenhagen S, Denmark. E-mail: dh@ssi.dk

{gamma}-Aminobutyric acid (GABA) is a neurotransmitter that also occurs in a few non-neuronal cell types, where it may serve as a paracrine modulator. GABA is biosynthesized from glutamate by glutamate decarboxylase (GAD) and from putrescine via diamine oxidase (DAO). GAD is demonstrable in several GABA-positive cell types but is undetectable in the GABA-containing gastrin cells and somatostatin cells of the antropyloric mucosa of the stomach. Using two antisera raised against synthetic peptides corresponding to two different regions of rat DAO, we now demonstrate strong reactivity for DAO in gastrin-positive cells of the rat antropyloric mucosa, whereas somatostatin-positive cells as well as other structures of the antrum are unreactive. Western blotting analysis of antrum and colon demonstrate that both antisera react with a single band of 85 kD, consistent with the predicted molecular weight of DAO. Expression of DAO mRNA in the antrum is demonstrated by reverse transcriptase polymerase chain reaction (RT-PCR). Our results strongly indicate that gastrin cells produce GABA via DAO-catalyzed oxidation of putrescine, and experimental data moreover suggest that the biosynthesis of GABA is regulated by the prandial state. Because GABA modulates release of somatostatin, these results point to a new mechanism of paracrine interaction between gastrin cells and somatostatin cells.

(J Histochem Cytochem 48:839–846, 2000)

Key Words: diamine oxidase, DAO, GABA, polyamines, putrescine, gastrin, antrum, immunohistochemistry, paracrine interaction


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