Residual Glucose Taste in T1R3 Knockout but not TRPM5 Knockout Mice.

Knockout (KO) mice missing the sweet taste receptor subunit T1R3 or the signaling protein TRPM5 have greatly attenuated sweetener preferences. Yet both types of KO mice develop preferences for glucose but not fructose in 24-h tests, which has been attributed to the postoral reinforcing actions of glucose. Here we probed for residual sugar taste sensitivity in KO mice. Unlike wildtype (WT) mice, food-restricted T1R3 KO and TRPM5 KO mice displayed little attraction for 8% glucose and 8% fructose in 1-min, two-bottle choice tests. However, in 1-h tests about half of the T1R3 KO mice displayed a significant preference for glucose over fructose (78-84%), while WT mice showed either no or weak preferences (41-56%) for glucose. Following one-bottle training sessions, WT mice display greater glucose preferences although still weaker than those observed in T1R3 KO mice. In contrast, TRPM5 KO mice were indifferent to sugars in 1-h tests but developed a strong preference for glucose over fructose in 24-h tests. T1R3 taste cells contain the sodium glucose cotransporter 1 (SGLT1) and the ATP-gated K+ (KATP) metabolic sensor, which may mediate the unlearned glucose preference displayed by T1R3 KO mice. Unlike WT mice, many T1R3 KO mice strongly preferred glucose to a non-metabolizable glucose analog (α-methyl-D-glucopyranoside, MDG) in initial 1-h choice tests. Glucose and MDG are both ligands for SGLT1 which indicates that SGLT1 sensing does not mediate the glucose preference of T1R3 KO mice. Instead, KATP sensing and/or other oral sensors are implicated. The MDG findings also argue against postoral sensing as the primary source of the initial glucose preference displayed by T1R3 KO mice. Why only half of the T1R3 KO mice showed this preference in 1-h tests remains to be determined. All T1R3 KO mice preferred glucose to fructose in 24-h tests, which appears to be due to both oral and postoral glucose sensing.

Sclafani A ，Zukerman S ，Ackroff K 《-》

Impact of T1r3 and Trpm5 on carbohydrate preference and acceptance in C57BL/6 mice.

Zukerman S ，Glendinning JI ，Margolskee RF ，Sclafani A ... -  《-》

Flavor preference conditioning by different sugars in sweet ageusic Trpm5 knockout mice.

Knockout (KO) mice missing the taste signaling protein Trpm5 have greatly attenuated sweetener preferences but develop strong preferences for glucose in 24-h tests, which is attributed to post-oral sugar conditioning. Trpm5 KO mice express mild preferences for galactose but no preferences for fructose in 24-h tests, which suggests that these sugars differ in their post-oral reinforcing effects. Here we investigated sugar-conditioned flavor preferences in Trpm5 KO and C57BL/6J wildtype (B6) mice. The mice were trained to consume a flavored (CS+, e.g. grape) 8% sugar solution and flavored (CS-, e.g., cherry) water on alternating days followed by two-bottle choice tests with CS+ vs. CS- flavors in water and with unflavored sugar vs. water. The KO mice displayed strong preferences (>80%) for the CS+ glucose and CS+ galactose but not for the CS+ fructose flavor. They also preferred glucose and galactose, but not fructose to water. In contrast, the B6 mice preferred all three CS+ flavors to the CS- flavor, and all three sugars to water. In tests with the non-metabolizable sugar α-methyl-d-glucopyranoside (MDG), the KO and B6 mice preferred 8% MDG to water but did not prefer the CS+ 8% MDG to CS-. However, they preferred a CS+ flavor mixed with 4% MDG over the CS- flavor. Trpm5 KO mice also preferred galactose and MDG to fructose in direct choice tests. The Trpm5 KO data indicate that glucose and, to a lesser extent, galactose and MDG have post-oral reinforcing actions that stimulate intake and preference while fructose has a much weaker effect. The CS+ flavor and sugar preferences of B6 mice may be mediated by the sweet taste and/or post-oral actions of the various sugars. Glucose, galactose, and MDG, but not fructose, are ligands for the sodium-glucose transporter 1 (SGLT1) which is implicated in post-oral sugar conditioning in B6 mice.

Sclafani A ，Ackroff K 《-》

Fat preference deficits and experience-induced recovery in global taste-deficient Trpm5 and Calhm1 knockout mice.

There is much evidence that gustation mediates the preference for dietary fat in rodents. Several studies indicate that mice have fat taste receptors that activate downstream signaling elements, including TRPM5 and CALHM1 ion channels and P2×2/P2×3 purinergic gustatory nerve receptors. Experiment 1 further documented the involvement of TRPM5 in fat appetite by giving Trpm5 knockout (KO) mice, which show global taste deficits, 24-h two-bottle choice tests with ascending concentrations of soybean oil (0.1 - 10% Intralipid) vs. water. Unlike wildtype (WT) mice, naive Trpm5 KO mice were indifferent to 0.5 - 2.5% fat. They preferred 5-10% fat but consumed much less than WT mice. The same KO mice preferred all fat concentrations in a second test series, which is attributed to a postoral fat conditioned attraction to the non-taste flavor qualities of the Intralipid, although they consumed less fat than the WT mice. The fat preference deficits of the Trpm5 KO mice were as great or greater than those observed in Calhm1 KO mice, another KO line with global taste deficits. Experiment 2 examined experience-enhanced fat preferences in Trpm5 KO and Calhm1 KO mice by giving them one-bottle training with 1%, 2.5%, and 5% fat prior to two-bottle fat vs. water tests. The KO mice displayed increased two-bottle preferences for all concentrations, although they still consumed less 1% and 2.5% fat than WT mice. Thus, the postoral actions of fat induce robust preferences for fat in taste-deficient mice, but do not stimulate the high fat intakes observed in WT mice with normal fat taste signaling.

Sclafani A ，Ackroff K 《-》

Sucrose-conditioned flavor preferences in sweet ageusic T1r3 and Calhm1 knockout mice.

The present study compared the ability of sweet ageusic T1r3 knockout (KO) and Calhm1 KO mice to acquire preferences for a sucrose-paired flavor as well as for unflavored sucrose. The KO and wildtype (WT) mice were given 24-h one-bottle access to 8% sucrose containing one flavor CS+, e.g., grape) and to water containing a different flavor (CS-, e.g., cherry) over 4 training days. In subsequent two-bottle tests with the flavors in water only, the T1r3 KO and Calhm1 KO mice, like WT mice, preferred the CS+ to the CS-. After training with flavored solutions, both KO groups also preferred unflavored 8% sucrose to water although Calhm1 KO mice required more sugar experience to match the preference of the T1r3 KO mice. These findings demonstrate that Calhm1 KO mice, like T1r3 KO mice and WT mice, are sensitive to the post-oral preference conditioning actions of sucrose and can discriminate sugar from water. Yet, despite their acquired sucrose preferences, the Calhm1 KO and T1r3 KO mice consumed only half as much sugar per day as did WT mice. Thus, sweet taste signaling elements are not needed in the gut for sugar conditioning, but sweet taste signaling in the mouth is essential for the full expression of sugar appetite.

Sclafani A ，Marambaud P ，Ackroff K 《-》