Proton currents through amiloride-sensitive Na channels in hamster taste cells. Role in acid transduction.
The activity of taste cells maintained in the intact hamster tongue was monitored in response to acid stimulation by recording action currents from taste receptor cells with an extracellular "macro" patch pipette: a glass pipette was pressed over the taste pore of fungiform papillae and perfused with citric acid, hydrochloric acid, or NaCl. Because this technique restricted stimulus application to the small surface area of the apical membranes of the taste cells, many nonspecific, and potentially detrimental, effects of acid stimulation could be avoided. Acid stimulation reliably elicited fast transient currents (action currents of average amplitude, 9 pA) which were consistently smaller than those elicited by NaCl (29 pA). The frequency of action currents elicited by acid stimuli increased in a dose-dependent manner with decreasing pH from a threshold of about pH 5.0. Acid-elicited responses were independent of K+, Na+, Cl-, or Ca2+ at physiological (salivary) concentrations, and were unaffected by anthracene-9-carboxylic acid, tetraethylammonium bromide, diisothiocyanate-stilbene-2,2'-disulfonic acid, vanadate, or Cd2+. In contrast, amiloride (< or = 30 microM) fully and reversibly suppressed acid-evoked action currents. At submaximal amiloride concentrations, the frequency and amplitude of the action currents were reduced, indicating a reduction of the taste cell apical conductance concomitant with a decrease in cell excitation. Exposure to low pH elicited, in addition to transient currents, an amiloride-sensitive sustained d.c. current. This current is apparently carried by protons instead of Na+ through amiloride-sensitive channels. When citric acid was applied while the taste bud was stimulated by NaCl, the action currents became smaller and the response resembled that produced by acid alone. Because of the strong interdependence of the acid and salt (NaCl) responses when both stimuli are applied simultaneously, and because of the similarity in the concentration dependence of amiloride block, we conclude that amiloride-sensitive Na+ channels on hamster taste receptor cells are permeable to protons and may play a role in acid (sour) taste.
Gilbertson
T A
TA
Department of Anatomy and Neurobiology, Colorado State University, Fort Collins 80523.
Avenet
P
P
Kinnamon
S C
SC
Roper
S D
SD
D000200:Action Potentials / Q000187:drug effects; D000584:Amiloride / Q000494:pharmacology; D000818:Animals; D002951:Citrates / Q000494:pharmacology; D019343:Citric Acid; D006224:Cricetinae; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D009092:Mucous Membrane / Q000187:drug effects / Q000378:metabolism; D010539:Permeability; D011522:Protons; D012464:Saliva, Artificial; D015222:Sodium Channels / Q000187:drug effects / Q000378:metabolism*; D012965:Sodium Chloride / Q000494:pharmacology; D013650:Taste Buds / Q000187:drug effects / Q000378:metabolism*
D002951:Citrates; D011522:Protons; D012464:Saliva, Artificial; D015222:Sodium Channels; D019343:Citric Acid; D012965:Sodium Chloride; D000584:Amiloride