Abstract

A plethora of molecular and functional studies in tetrapods has led to the discovery of multiple taste 1 receptor (T1R) genes encoding G-protein coupled receptors (GPCRs) responsible for sweet (T1R2 + T1R3) and umami (T1R1 + T1R3) taste. In fish, the T1R gene family repertoires greatly expanded because of several T1R2 gene duplications, and recent studies have shown T1R2 functional divergence from canonical mammalian sweet taste perceptions, putatively as an adaptive mechanism to develop distinct feeding strategies in highly diverse aquatic habitats. We addressed this question in the carnivore fish gilthead seabream (Sparus aurata), a model species of aquaculture interest, and found that the saT1R gene repertoire consists of eight members including saT1R1, saT1R3 and six saT1R2a-f gene duplicates, adding further evidence to the evolutionary complexity of fishT1Rs families. To analyze saT1R taste functions, we first developed a stable gene reporter system based on Ca2+-dependent calcineurin/NFAT signaling to examine specifically in vitro the responses of a subset of saT1R heterodimers to L-amino acids (L-AAs) and sweet ligands. We show that although differentially tuned in sensitivity and magnitude of responses, saT1R1/R3, saT1R2a/R3 and saT1R2b/R3 may equally serve to transduce amino acid taste sensations. Furthermore, we present preliminary information on the potential involvement of the Gi protein alpha subunits saGαi1 and saGαi2 in taste signal transduction.

Highlights

  • Perception of sweet, umami and bitter taste is mediated by two distinct G protein-coupled receptor (GPCR) families, namely by taste 1 receptor (T1R) and T2R, mostly expressed in taste buds [1,2,3]

  • Five saT1R genes were initially identified through blast searches of the preliminary seabream genome assembly, subsequently cloned in their complete coding sequences [saT1R1 (846 AAs, XP_030277517), saT1R2a (826 AAs, XP_030278006), saT1R2b (824 AAs, XP_030278002), saT1R2c (817 AAs, MT892735), saT1R3 (856 AAs, XP_030274769)], and used in the pharmacological characterization of different receptor combinations

  • Information available up to date indicates that the saT1R gene repertoire comprises eight members: two one-to-one orthologs of T1R1 and T1R3 vertebrate genes and six saT1R2 duplicate members highly conserved to each other, and moderately with respect to saT1R1 and saT1R3 (± 47% and 42% respectively)

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Summary

Introduction

Perception of sweet, umami and bitter taste is mediated by two distinct G protein-coupled receptor (GPCR) families, namely by taste 1 receptor (T1R) and T2R, mostly expressed in taste buds [1,2,3]. The T1R gene repertoire is relatively conserved and consists of three ancient duplicated genes that diverged before fish/tetrapod diversification, about 400 million years ago (MYA) [4,5]. The. T1R family’s receptor subunits (T1R1, T1R2 and T1R3) dimerize to form functional receptors. The combination of T1R1/T1R3 forms the umami receptor and is activated by amino and nucleic acids, and the T1R2/T1R3 heterodimer responds to natural and artificial sugars including sweet proteins and.

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