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Identification and characterization of the bovine G protein-coupled receptor GPR41 and GPR43 genes¹

A. Wang^*, Z. Gu^*,2, B. Heid^*, R. M. Akers and H. Jiang^*,3

^* Department of Animal and Poultry Sciences, and
Department of Dairy Science, Virginia Polytechnic Institute and State University, Blacksburg 24061

³ Corresponding author: hojiang{at}vt.edu

Volatile fatty acids (VFA), including acetate, propionate, and butyrate, are not only a primary source of energy, but also regulate rumen development, insulin and glucagon secretion, and other physiological processes in cattle and sheep. The mechanism underlying the regulatory effects of VFA is unknown. Recent "reverse pharmacology" studies identified human G protein-coupled receptors GPR41 and GPR43 as receptors for short-chain fatty acids. It is possible that proteins similar to human GPR41 and GPR43 mediate the regulatory effects of VFA in cattle. In this study, we determined first, whether the bovine genome contains genes similar to the human GPR41 and GPR43 genes; second, whether and where these genes are expressed in cattle; and third, if the proteins encoded by these genes can be activated by acetate, propionate, and butyrate. A search of GenBank revealed bovine genomic sequences and expressed sequence tags highly similar to the human GPR41 and GPR43 DNA and cDNA sequences. The protein-coding and 5' untranslated regions of the bovine GPR41 and GPR43 mRNA were cloned and sequenced from spleen tissue. Based on these sequences, the bovine GPR41 gene contains 3 exons and its transcription is initiated at 2 leader exons, generating 2 GPR41 mRNA variants differing in the 5' untranslated region. The bovine GPR43 gene contains 2 exons and transcription of this gene is initiated from a single start site. The amino acid sequences deduced from the bovine GPR41 and GPR43 mRNA sequences are more than 75% identical to those of the human GPR41 and GPR43 and are predicted to encode 7 transmembrane domains, typical of G protein-coupled receptors. Both bovine GPR41 and GPR43 mRNA were detected in a variety of tissues including rumen and pancreas. In a cell system, interaction of the overexpressed bovine GPR41 or GPR43 protein with acetate, propionate, or butyrate inhibited luciferase reporter expression from a cyclic AMP-responsive promoter, suggesting that the bovine GPR41 and GPR43 proteins couple to G_i/11. In total, these results demonstrate that the bovine genome encodes functional GPR41 and GPR43 genes and suggest that GPR41 and GPR43 may play a role in the regulatory effects of VFA in cattle.

Key Words: G protein-coupled receptors GPR41 and GPR43 • volatile fatty acids • cattle

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