Lucero M T, Pappone P A
Department of Animal Physiology, University of California, Davis 95616.
J Gen Physiol. 1989 Mar;93(3):451-72. doi: 10.1085/jgp.93.3.451.
We studied the membrane currents of isolated cultured brown fat cells from neonatal rats using whole-cell and single-channel voltage-clamp recording. All brown fat cells that were recorded from had voltage-gated K currents as their predominant membrane current. No inward currents were seen in these experiments. The K currents of brown fat cells resemble the delayed rectifier currents of nerve and muscle cells. The channels were highly selective for K+, showing a 58-mV change in reversal potential for a 10-fold change in the external [K+]. Their selectivity was typical for K channels, with relative permeabilities of K+ greater than Rb+ greater than NH+4 much greater than Cs+, Na+. The K currents in brown adipocytes activated with a sigmoidal delay after depolarizations to membrane potentials positive to -50 mV. Activation was half maximal at a potential of -28 mV and did not require the presence of significant concentrations of internal calcium. Maximal voltage-activated K conductance averaged 20 nS in high external K+ solutions. The K currents inactivated slowly with sustained depolarization with time constants for the inactivation process on the order of hundreds of milliseconds to tens of seconds. The K channels had an average single-channel conductance of 9 pS and a channel density of approximately 1,000 channels/cell. The K current was blocked by tetraethylammonium or 4-aminopyridine with half maximal block occurring at concentrations of 1-2 mM for either blocker. K currents were unaffected by two blockers of Ca2+-activated K channels, charybdotoxin and apamin. Bath-applied norepinephrine did not affect the K currents or other membrane currents under our experimental conditions. These properties of the K channels indicate that they could produce an increase in the K+ permeability of the brown fat cell membrane during the depolarization that accompanies norepinephrine-stimulated thermogenesis, but that they do not contribute directly to the norepinephrine-induced depolarization.
我们使用全细胞和单通道电压钳记录技术,研究了新生大鼠分离培养的棕色脂肪细胞的膜电流。所有记录到的棕色脂肪细胞都以电压门控钾电流作为其主要膜电流。在这些实验中未观察到内向电流。棕色脂肪细胞的钾电流类似于神经和肌肉细胞的延迟整流电流。这些通道对钾离子具有高度选择性,细胞外[K⁺]变化10倍时,反转电位变化58 mV。它们的选择性是钾通道的典型特征,钾离子的相对通透性大于铷离子大于铵离子远大于铯离子、钠离子。棕色脂肪细胞中的钾电流在去极化至膜电位高于 -50 mV后呈S形延迟激活。在 -28 mV电位时激活达到半数最大值,且不需要存在显著浓度的细胞内钙。在高细胞外钾离子溶液中,最大电压激活钾电导平均为20 nS。钾电流在持续去极化时缓慢失活,失活过程的时间常数在数百毫秒到数十秒的范围内。钾通道的平均单通道电导为9 pS,通道密度约为1000个通道/细胞。钾电流被四乙铵或4 - 氨基吡啶阻断,两种阻断剂半数最大阻断浓度均为1 - 2 mM。钾电流不受两种钙激活钾通道阻断剂(蝎毒素和蜂毒明肽)的影响。在我们的实验条件下,浴加去甲肾上腺素不影响钾电流或其他膜电流。钾通道的这些特性表明,它们可能在去甲肾上腺素刺激产热伴随的去极化过程中,使棕色脂肪细胞膜的钾离子通透性增加,但它们并不直接导致去甲肾上腺素诱导的去极化。