Béhé P, DeSimone J A, Avenet P, Lindemann B
Department of Physiology, Universität des Saarlandes, Homburg/Saar, Federal Republic of Germany.
J Gen Physiol. 1990 Nov;96(5):1061-84. doi: 10.1085/jgp.96.5.1061.
Taste buds were isolated from the fungiform papilla of the rat tongue and the receptor cells (TRCs) were patch clamped. Seals were obtained on the basolateral membrane of 281 TRCs, protruding from the intact taste buds or isolated by micro-dissection. In whole-cell configuration 72% of the cells had a TTX blockable transient Na inward current (mean peak amplitude 0.74 nA). All cells had outward K currents. Their activation was slower than for the Na current and a slow inactivation was also noticeable. The K currents were blocked by tetraethylammonium, Ba, and 4-aminopyridine, and were absent when the pipette contained Cs instead of K. With 100 mM Ba or 100 mM Ca in the bath, two types of inward current were observed. An L-type Ca current (ICaL) activated at -20 mV had a mean peak amplitude of 440 pA and inactivated very slowly. At 3 mM Ca the activation threshold of ICaL was near -40 mV. A transient T-type current (ICaT) activated at -50 mV had an average peak amplitude of 53 pA and inactivated with a time constant of 36 ms at -30 mV. ICaL was blocked more efficiently by Cd and D600 than ICaT. ICaT was blocked by 0.2 mM Ni and half blocked by 200 microM amiloride. In whole-cell voltage clamp, Na-saccharin caused (in 34% of 55 cells tested) a decrease in outward K currents by 21%, which may be expected to depolarize the TRCs. Also, Na-saccharin caused some taste cells to fire action potentials (on-cell, 7 out of 24 cells; whole-cell, 2 out of 38 cells responding to saccharin) of amplitudes sufficient to activate ICaL. Thus the action potentials will cause Ca inflow, which may trigger release of transmitter.
从大鼠舌的菌状乳头分离出味蕾,并对受体细胞(TRCs)进行膜片钳记录。在281个从完整味蕾突出或通过显微切割分离的TRCs的基底外侧膜上获得了封接。在全细胞模式下,72%的细胞具有可被河豚毒素阻断的瞬时内向钠电流(平均峰值幅度为0.74 nA)。所有细胞都有外向钾电流。它们的激活比钠电流慢,并且缓慢失活也很明显。钾电流被四乙铵、钡和4-氨基吡啶阻断,当移液管中含有铯而不是钾时钾电流消失。在浴液中加入100 mM钡或100 mM钙时,观察到两种内向电流。一种L型钙电流(ICaL)在-20 mV激活,平均峰值幅度为440 pA,失活非常缓慢。在3 mM钙时,ICaL的激活阈值接近-40 mV。一种瞬时T型电流(ICaT)在-50 mV激活,平均峰值幅度为53 pA,在-30 mV时以36 ms的时间常数失活。ICaL比ICaT更有效地被镉和D600阻断。ICaT被0.2 mM镍阻断,被200 microM氨氯吡咪半阻断。在全细胞电压钳记录中,钠-糖精(在测试的55个细胞中的34%)使外向钾电流降低了21%,这可能会使TRCs去极化。此外,钠-糖精使一些味觉细胞产生动作电位(在细胞贴附模式下,24个细胞中有7个;在全细胞模式下,38个对糖精有反应的细胞中有2个),其幅度足以激活ICaL。因此,动作电位将导致钙内流,这可能触发递质释放。