Alpern R J
J Clin Invest. 1987 Apr;79(4):1026-30. doi: 10.1172/JCI112914.
To examine whether Cl/base exchange is present on the apical membrane of the proximal convoluted tubule, cell pH was measured fluorometrically in the in vivo microperfused rat proximal tubule with (2',7')-bis(carboxyethyl)-(5,6)-carboxyfluorescein. The effect of luminal chloride addition was examined in tubules perfused symmetrically with chloride-free solutions. In the absence of inhibitors, luminal chloride addition did not affect cell pH. However, after inhibition of basolateral membrane anion transport with peritubular 4-acetamido-4'-isothiocyano-(2,2')-disulfonic-stilbene (to amplify effects of apical membrane transport on cell pH), luminal chloride addition caused a small cell acidification (delta pHi = 0.02). When 1 mM formate was added to the solutions, luminal chloride addition caused a larger change in cell pH (delta pHi = 0.06) that was inhibited by (4,4')-diisothiocyano-(2,2')-disulfonic-stilbene. This stimulation of Cl/base exchange was not seen with 1 mM acetate addition. These results demonstrate apical membrane Cl/base exchange, a significant fraction of which is dependent on the presence of formate and probably represents Cl/formate exchange.
为了检测近端小管顶端膜上是否存在氯/碱基交换,使用(2',7')-双(羧乙基)-(5,6)-羧基荧光素通过荧光法在体内微灌注大鼠近端小管中测量细胞pH。在无氯溶液对称灌注的小管中检测管腔添加氯的影响。在没有抑制剂的情况下,管腔添加氯不影响细胞pH。然而,在用肾小管周围的4-乙酰氨基-4'-异硫氰基-(2,2')-二磺酸芪抑制基底外侧膜阴离子转运后(以放大顶端膜转运对细胞pH的影响),管腔添加氯导致细胞轻微酸化(ΔpHi = 0.02)。当向溶液中添加1 mM甲酸盐时,管腔添加氯导致细胞pH发生更大变化(ΔpHi = 0.06),这种变化被(4,4')-二异硫氰基-(2,2')-二磺酸芪抑制。添加1 mM乙酸盐时未观察到这种对氯/碱基交换的刺激作用。这些结果证明了顶端膜存在氯/碱基交换,其中很大一部分依赖于甲酸盐的存在,可能代表氯/甲酸盐交换。