Oelberg D G, Xu F
Department of Pediatrics, Eastern Virginia Medical School, Norfolk, Virginia, 23510, USA.
Mol Genet Metab. 1998 Nov;65(3):220-8. doi: 10.1006/mgme.1998.2735.
Choline is an important substrate in alveolar epithelia for both surfactant production and cellular maintenance. The underlying mechanisms of uptake and sites of membrane transport remain uncertain. To test the hypothesis that choline transport occurs at the basolateral side of alveolar epithelia by both Na+-independent and -dependent mechanisms, plasma membrane vesicles were prepared from the apical and basolateral membranes of mature porcine type II pneumocytes. Choline+ transport was assayed by uptake of [3H]choline+ by enriched apical or basolateral vesicles. In the presence of imposed, inside-negative charge gradients, basolateral vesicles exhibited early overshoot of [3H]choline+ uptake unaffected by the presence or absence of external Na+ (541 +/- 53 vs 564 +/- 79 pmol/mg protein (NS)). High sensitivity to hemicholinium-3 was observed in the presence or absence of Na+. In the absence of inside-negative charge gradients, uptake was reduced 12-fold in the presence or absence of Na+, and external choline+ induced internal alkalization of acidified basolateral vesicles. Accumulative [3H]choline+ uptakes by apical vesicles in the presence or absence of inside-negative charge gradients and Na+ were insignificant. We conclude that predominant choline+ uptake by type II pneumocytes occurs at the basolateral membrane by Na+-independent, electrogenic choline+ conductance. The presence of electroneutral choline+/H+ exchange is suggested.
胆碱是肺泡上皮细胞中用于表面活性剂产生和细胞维持的重要底物。其摄取的潜在机制和膜转运位点仍不确定。为了验证胆碱转运通过不依赖钠和依赖钠的机制在肺泡上皮细胞的基底外侧发生这一假设,从成熟猪II型肺细胞的顶端和基底外侧膜制备了质膜囊泡。通过富集的顶端或基底外侧囊泡摄取[3H]胆碱+来测定胆碱+转运。在施加的内负电荷梯度存在的情况下,基底外侧囊泡表现出[3H]胆碱+摄取的早期过冲,不受外部钠存在与否的影响(541±53对564±79 pmol/mg蛋白质(无显著性差异))。在有或没有钠的情况下,均观察到对hemicholinium-3的高敏感性。在没有内负电荷梯度的情况下,无论有无钠,摄取均降低12倍,并且外部胆碱+诱导酸化的基底外侧囊泡内部碱化。在有或没有内负电荷梯度和钠的情况下,顶端囊泡的累积[3H]胆碱+摄取不显著。我们得出结论,II型肺细胞主要通过不依赖钠的、电生性胆碱+电导在基底外侧膜摄取胆碱+。提示存在电中性胆碱+/H+交换。