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

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.

Smith L ，Moran AW ，Al-Rammahi M ，Daly K ，Shirazi-Beechey SP ... -  《Frontiers in Veterinary Science》

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

The perception of sweet is mediated by the sweet taste receptor T1R2-T1R3 expressed in taste cells of the lingual epithelium. This receptor is also expressed in intestinal enteroendocrine cells and is required for sensing luminal sugars and sweeteners to regulate expression of intestinal Na+-glucose cotransporter 1 (SGLT1). There are some notable differences amongst species in the ability to detect certain non-nutritive (artificial) sweeteners. Amino acid substitutions and pseudogenization of taste receptor genes are responsible for these disparities. Using heterologous expression, we demonstrate that the commonly used non-nutritive sweeteners sucralose, saccharin and acesulfame K activate pig T1R2-T1R3, but that aspartame and cyclamate do not. Furthermore, we show that in vitro sweetener activation of pig T1R2-T1R3 mirrors the sweetener stimulation of the gut-expressed receptor in vivo. Considering that sweeteners are included in animal feed worldwide, determination of taste receptor specificities in different species is essential for the development of scientifically-based dietary formulations.

Daly K ，Moran AW ，Al-Rammahi M ，Weatherburn D ，Shirazi-Beechey SP ... -  《-》

Sweet taste receptor expression in ruminant intestine and its activation by artificial sweeteners to regulate glucose absorption.

Absorption of glucose from the lumen of the intestine into enterocytes is accomplished by sodium-glucose co-transporter 1 (SGLT1). In the majority of mammalian species, expression (this includes activity) of SGLT1 is upregulated in response to increased dietary monosaccharides. This regulatory pathway is initiated by sensing of luminal sugar by the gut-expressed sweet taste receptor. The objectives of our studies were to determine (1) if the ruminant intestine expresses the sweet taste receptor, which consists of two subunits [taste 1 receptor 2 (T1R2) and 3 (T1R3)], and other key signaling molecules required for SGLT1 upregulation in nonruminant intestines, and (2) whether T1R2-T1R3 sensing of artificial sweeteners induces release of glucagon-like peptide-2 (GLP-2) and enhances SGLT1 expression. We found that the small intestine of sheep and cattle express T1R2, T1R3, G-protein gustducin, and GLP-2 in enteroendocrine L-cells. Maintaining 110-d-old ruminating calves for 60d on a diet containing a starter concentrate and the artificial sweetener Sucram (consisting of saccharin and neohesperidin dihydrochalcone; Pancosma SA, Geneva, Switzerland) enhances (1) Na(+)-dependent d-glucose uptake by over 3-fold, (2) villus height and crypt depth by 1.4- and 1.2-fold, and (3) maltase- and alkaline phosphatase-specific activity by 1.5-fold compared to calves maintained on the same diet without Sucram. No statistically significant differences were observed for rates of intestinal glucose uptake, villus height, crypt depth, or enzyme activities between 50-d-old milk-fed calves and calves maintained on the same diet containing Sucram. When adult cows were kept on a diet containing 80:20 ryegrass hay-to-concentrate supplemented with Sucram, more than a 7-fold increase in SGLT1 protein abundance was noted. Collectively, the data indicate that inclusion of this artificial sweetener enhances SGLT1 expression and mucosal growth in ruminant animals. Exposure of ruminant sheep intestinal segments to saccharin or neohesperidin dihydrochalcone evokes secretion of GLP-2, the gut hormone known to enhance intestinal glucose absorption and mucosal growth. Artificial sweeteners, such as Sucram, at small concentrations are potent activators of T1R2-T1R3 (600-fold>glucose). This, combined with oral bioavailability of T1R2-T1R3 and the understanding that artificial sweetener-induced receptor activation evokes GLP-2 release (thus leading to increased SGLT1 expression and mucosal growth), make this receptor a suitable target for dietary manipulation.

Moran AW ，Al-Rammahi M ，Zhang C ，Bravo D ，Calsamiglia S ，Shirazi-Beechey SP ... -  《-》

Expression of sweet receptor components in equine small intestine: relevance to intestinal glucose transport.

Daly K ，Al-Rammahi M ，Arora DK ，Moran AW ，Proudman CJ ，Ninomiya Y ，Shirazi-Beechey SP ... -  《-》

Expression of Na+/glucose co-transporter 1 (SGLT1) is enhanced by supplementation of the diet of weaning piglets with artificial sweeteners.

Moran AW ，Al-Rammahi MA ，Arora DK ，Batchelor DJ ，Coulter EA ，Daly K ，Ionescu C ，Bravo D ，Shirazi-Beechey SP ... -  《-》