Barakat L, Wang D, Bordey A
Department of Neurosurgery, Yale University, New Haven, CT 06520-8082, USA.
J Physiol. 2002 Jun 15;541(Pt 3):753-67. doi: 10.1113/jphysiol.2001.015834.
Taurine uptake is essential for the maintenance of millimolar intracellular concentrations of taurine, which is released during ischaemia and is thought to be neuroprotective. To determine whether Bergmann glia express functional transporters that can mediate both taurine uptake and efflux, whole-cell patch-clamp recordings were obtained from these cells in rat cerebellar slices. Taurine-induced inward currents can be pharmacologically separated into GABA(A) receptor and taurine transporter currents. In the presence of GABA receptor blockers, residual taurine currents averaged -28 pA at -70 mV and were strictly inwardly rectifying between -70 and +50 mV. These residual currents were also abolished by external Na+ removal and diminished by reduction of external Cl-, consistent with transport currents. Taurine transport currents were reduced by a taurine transporter inhibitor, guanidinoethyl sulphonate (GES). Other classical inhibitors reduced taurine transport currents with an order of potency (hypotaurine > beta-alanine > GES > GABA) similar to that reported for cloned rat taurine transporters. Following intracellular taurine perfusion during the recording, a progressively developing outward current could be observed at -50 mV but not at -70 mV. Intracellular perfusion of taurine also decreased taurine-induced inward currents at both holding potentials. Outward currents induced by intracellular taurine increased in amplitude with depolarization, activated near -50 mV, and were affected by GES. For the first time, these results demonstrate that taurine activates both GABA(A) receptors and Na+/Cl--dependent taurine transporters in Bergmann glia in slices. In addition, our data show that taurine transporters can work in reverse and can probably mediate taurine efflux under ischaemic conditions.
牛磺酸摄取对于维持细胞内毫摩尔浓度的牛磺酸至关重要,牛磺酸在缺血时释放,被认为具有神经保护作用。为了确定伯格曼胶质细胞是否表达能够介导牛磺酸摄取和外流的功能性转运体,在大鼠小脑切片的这些细胞上进行了全细胞膜片钳记录。牛磺酸诱导的内向电流在药理学上可分为GABA(A)受体电流和牛磺酸转运体电流。在存在GABA受体阻滞剂的情况下,残余牛磺酸电流在-70 mV时平均为-28 pA,并且在-70至+50 mV之间严格内向整流。这些残余电流也因去除细胞外Na+而被消除,并因降低细胞外Cl-而减小,这与转运电流一致。牛磺酸转运体抑制剂胍基乙磺酸盐(GES)可降低牛磺酸转运电流。其他经典抑制剂降低牛磺酸转运电流的效力顺序(次牛磺酸>β-丙氨酸>GES>GABA)与报道的克隆大鼠牛磺酸转运体相似。在记录过程中进行细胞内牛磺酸灌注后,在-50 mV但不在-70 mV处可观察到逐渐发展的外向电流。细胞内灌注牛磺酸也降低了两个钳制电位下牛磺酸诱导的内向电流。细胞内牛磺酸诱导的外向电流幅度随去极化增加,在接近-50 mV时激活,并受GES影响。这些结果首次证明,牛磺酸在切片中的伯格曼胶质细胞中激活GABA(A)受体和Na+/Cl-依赖性牛磺酸转运体。此外,我们的数据表明,牛磺酸转运体可以反向工作,并且可能在缺血条件下介导牛磺酸外流。