Non-nutritive sweetener activation of the pig sweet taste receptor T1R2-T1R3 in vitro mirrors sweetener stimulation of the gut-expressed receptor in vivo

Daly, Kristian; Moran, Andrew W.; Al-Rammahi, Miran; Weatherburn, Darren; Shirazi-Beechey, Soraya P.

doi:10.1016/j.bbrc.2021.01.032

Cited by 6 publications

(8 citation statements)

References 24 publications

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“…Immunocytochemistry was performed as described before ( 12 ). Briefly, human embryonic kidney (HEK)-293PEAKrapid cells that stably express the chimeric G-protein subunit, Gα15, were transiently transfected with the horse sweet receptor constructs (Tas1r2pcDNA3/TO and Tas1r3pcDNA5/FRT/PM) or with an empty pcDNA3/TO vector (negative control) using Lipofectamine2000 according to manufacturer’s instruction.…”

Section: Methodsmentioning

confidence: 99%

“…For the assessment of horse T1R2-T1R3 function, HEK293PEAKrapid Gα15 cells, were seeded into poly-D-lysing coated 96-well plates and transfected with either horse sweet receptor constructs or empty vector (as described above under immunocytochemistry). After 48 h incubation, cells were loaded with Ca 2+ -sensitive Fluo-4 AM fluorescent dye as described previously ( 12 ). Sweeteners, sucralose, stevia 80, NHDC, cyclamate and the natural sugar, glucose were diluted to desired concentrations.…”

Section: Methodsmentioning

confidence: 99%

“…Ca 2+ responses of transfected cells upon sweetener application were measured using Flex Station3 microplate reader (Molecular Devices UK Ltd., Wokingham, United Kingdom). For each trace, sweetener was added 22 s after the start of scan, with fluorescent measurements (excitation 485 nm; emission 525 nm) taken every 1.4 for 500 s as described ( 12 ). Data were calculated from at least three set of experiments.…”

Section: Methodsmentioning

confidence: 99%

“…Signals were then normalized to background fluorescence (F). Plots of signal amplitude (ΔF/F) against concentration were used to calculate the EC 50 values as reported before ( 12 ). The EC 50 values of agonist-receptor interaction were calculated by nonlinear regression of amplitude plots to the function f(x) = 100/[1 + (EC 50 /x)nH], where x is the agonist concentration and nH is the Hill coefficient ( 15 ).…”

Section: Methodsmentioning

confidence: 99%

“…In this paper, we have assessed the ability of several sweeteners in activating horse T1R2-T1R3 using a heterologous expression system ( 12 ). We show here that sucralose, stevia 80 and NHDC, at much lower concentrations than glucose, activate horse T1R2-T1R3, whereas cyclamate does not.…”

Section: Introductionmentioning

confidence: 99%

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Determination of sweetener specificity of horse gut-expressed sweet taste receptor T1R2-T1R3 and its significance for energy provision and hydration

Smith,

Moran,

Al-Rammahi

et al. 2024

Front. Vet. Sci.

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Studies carried out in several species have demonstrated that detection of low-calorie sweeteners in the lumen of the intestine, by the sweet receptor, T1R2-T1R3, initiates a signaling pathway leading to enhanced expression and activity of intestinal Na+/glucose cotransporter 1, SGLT1. This results in an increased gut capacity to absorb glucose, sodium chloride and water, the basis for oral rehydration therapy. Horses express T1R2, T1R3 and downstream signaling elements in the intestinal tissue. As such, the potential of sweetener-stimulation of T1R2-T1R3 leading to upregulation of SGLT1 allows the provision of more glucose (energy) and hydration for horses. This is especially important when the need for glucose increases during strenuous exercise, pregnancy, and lactation. There are significant differences among species in the ability to detect sweeteners. Amino acid substitutions and pseudogenization of taste receptor genes underlie these variations. Nothing is known about the sweetener specificity of horse T1R2-T1R3. Using heterologous expression methodology, we demonstrate that sweeteners sucralose, stevia and neohesperidin dihydrochalcone (NHDC) activate horse T1R2-T1R3, but cyclamate does not. Determination of sweetener specificity of equine sweet receptor is crucial for developing suitable dietary additives to optimize glucose absorption, hydration and avoiding the intestinal disease brought about by microbial fermentation of unabsorbed carbohydrate reaching the large intestine.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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