Changes in IP3 and cytosolic Ca2+ in response to sugars and non-sugar sweeteners in transduction of sweet taste in the rat.
1. The transduction pathways of sweet-sensitive cells in rat circumvallate (CV) taste buds were investigated with assays for inositol 1,4,5-trisphosphate (IP3) and with Ca2+ imaging. Stimulation with the non-sugar sweeteners SC-45647 and saccharin rapidly increased the cellular content of IP3 by 400 pmol (mg protein)-1, while sucrose had a much smaller effect on IP3. As shown previously, sucrose, but not saccharin, increased the content of cyclic adenosine monophosphate (cAMP) of this preparation. 2. Stimulation of isolated CV taste buds with SC-45647 increased the cytosolic Ca2+ concentration ([Ca2+]i) by 56.7 +/- 3.2 nM (n = 181). Due to the non-confocality of the measuring system, these concentrations are underestimates. The increase in [Ca2+]i did not require the presence of extracellular Ca2+, suggesting that the Ca2+ release was from intracellular stores. 3. Individual cells responding to the non-sugar sweeteners with Ca2+ release also responded to sucrose and to forskolin with an increase in [Ca2+]i. Such cells did not respond to the bitter tastant denatonium chloride. 4. Responses to sucrose were abolished by lowering the Ca2+ concentration of the stimulus solution, indicating Ca2+ uptake from the extracellular medium. 5. The responses of sweet-sensitive cells to forskolin were also abolished when Ca2+ ions were omitted from the stimulus solution. They were partially inhibited by the presence of Co2+, Ni2+, D600 (methoxyverapamil) and amiloride, indicating multiple pathways of Ca2+ uptake activated by cAMP. 6. In conclusion, a sweet-sensitive cell of the rat responds to sucrose with an increase in cAMP and Ca2+ uptake, but to non-sugar sweeteners with an increase in IP3 and Ca2+ release. The increase in [Ca2+]i, common to both pathways, is presumably required for synaptic exocytosis and for signal termination.
Bernhardt
S J
SJ
Department of Biochemistry, Food Science and Nutrition, Hebrew University of Jerusalem, Israel.
Naim
M
M
Zehavi
U
U
Lindemann
B
B
D000584:Amiloride / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002241:Carbohydrates / Q000494:pharmacology; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000494:pharmacology; D005456:Fluorescent Dyes / Q000494:pharmacology; D016257:Fura-2 / Q000494:pharmacology; D005711:Gallopamil / Q000494:pharmacology; D007091:Image Processing, Computer-Assisted; D066298:In Vitro Techniques; D015544:Inositol 1,4,5-Trisphosphate / Q000378:metabolism*; D008297:Male; D008670:Metals / Q000494:pharmacology; D008856:Microscopy, Fluorescence; D000644:Quaternary Ammonium Compounds / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D012439:Saccharin / Q000494:pharmacology; D015398:Signal Transduction / Q000502:physiology*; D013395:Sucrose / Q000494:pharmacology; D013549:Sweetening Agents / Q000494:pharmacology; D013650:Taste Buds / Q000166:cytology / Q000378:metabolism*
D000584:Amiloride; D002118:Calcium; D002241:Carbohydrates; D005576:Colforsin; D000242:Cyclic AMP; D005456:Fluorescent Dyes; D016257:Fura-2; D005711:Gallopamil; D015544:Inositol 1,4,5-Trisphosphate; D008670:Metals; D000644:Quaternary Ammonium Compounds; D012439:Saccharin; D013395:Sucrose; D013549:Sweetening Agents; C043414:denatonium