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The Journal of Neuroscience, January 28, 2004, 24(4):938-946; doi:10.1523/JNEUROSCI.1374-03.2004

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Behavioral/Systems/Cognitive
Polymorphisms in the Taste Receptor Gene (Tas1r3) Region Are Associated with Saccharin Preference in 30 Mouse Strains

D. R. Reed,¹ S. Li,¹ X. Li,¹ L. Huang,¹ M. G. Tordoff,¹ R. Starling-Roney,¹ K. Taniguchi,^1,2 D. B. West,³ J. D. Ohmen,³ G. K. Beauchamp,^1,4 and A. A. Bachmanov¹

¹Monell Chemical Senses Center, Philadelphia, Pennsylvania 19104, ²Kitasato University, Aomori 034-8628, Japan, ³Pfizer Global Research and Development, Alameda, California 94502, and ⁴Department of Psychology and School of Veterinary Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104

The results of recent studies suggest that the mouse Sac (saccharin preference) locus is identical to the Tas1r3 (taste receptor) gene. The goal of this study was to identify Tas1r3 sequence variants associated with saccharin preference in a large number of inbred mouse strains. Initially, we sequenced 6.7 kb of the Tas1r3 gene and its flanking regions from six inbred mouse strains with high and low saccharin preference, including the strains in which the Sac alleles were described originally (C57BL/6J, Sac^b; DBA/2J, Sac^d). Of the 89 sequence variants detected among these six strains, eight polymorphic sites were significantly associated with preferences for 1.6 mM saccharin. Next, each of these eight variant sites were genotyped in 24 additional mouse strains. Analysis of the genotype–phenotype associations in all 30 strains showed the strongest association with saccharin preference at three sites: nucleotide (nt) –791 (3 bp insertion/deletion), nt +135 (Ser45Ser), and nt +179 (Ile60Thr). We measured Tas1r3 gene expression, transcript size, and T1R3 immunoreactivity in the taste tissue of two inbred mouse strains with different Tas1r3 haplotypes and saccharin preferences. The results of these experiments suggest that the polymorphisms associated with saccharin preference do not act by blocking gene expression, changing alternative splicing, or interfering with protein translation in taste tissue. The amino acid substitution (Ile60Thr) may influence the ability of the protein to form dimers or bind sweeteners. Here, we present data for future studies directed to experimentally confirm the function of these polymorphisms and highlight some of the difficulties of identifying specific DNA sequence variants that underlie quantitative trait loci.

Key words: sweet; preference; phenotype; genotype; haplotype; QTL

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Received May 7, 2003; revised November 3, 2003; accepted November 19, 2003.

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