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人胎盘滋养层细胞和绒毛膜癌细胞中质膜5-羟色胺转运体而非囊泡单胺转运体的功能性表达。

Functional expression of the plasma membrane serotonin transporter but not the vesicular monoamine transporter in human placental trophoblasts and choriocarcinoma cells.

作者信息

Prasad P D, Hoffmans B J, Moe A J, Smith C H, Leibach F H, Ganapathy V

机构信息

Department of Biochemistry and Molecular Biology, Medical College of Georgia, Augusta 30912-2100, USA.

出版信息

Placenta. 1996 May;17(4):201-7. doi: 10.1016/s0143-4004(96)90039-9.

Abstract

We investigated the functional expression of the plasma membrane serotonin transporter and the vesicular monoamine transporter in choriocarcinoma cells and normal trophoblasts. The RBL 2H3 cells, a rat basophilic leukaemia cell line, which express both transporters were used for comparison. The choriocarcinoma cells JAr and BeWo were found to possess the plasma membrane serotonin transporter as assessed by the presence of serotonin transport activity in intact cells that was Na(+)-dependent and was sensitive to inhibition by tricyclic and non-tricyclic antidepressants. The activity of the vesicular monoamine transporter in these cells was determined by measuring serotonin transport in digitonin-permeabilized cells. The transport in permeabilized cells was very slow, was not stimulated by ATP and was insensitive to inhibition by reserpine. Under similar conditions, the vesicular monoamine transporter activity was demonstrable in RBL cells, which was stimulated by ATP and was inhibitable by reserpine, bafilomycin A1 (an inhibitor of the V-type H(+)-pump) and carbonyl cyanide p-trifluoromethoxy phenylhydrazone (a protonophore which dissipates transmembrane H+ gradients). In corroboration with these findings, mRNA transcripts hybridizable to the vesicular monoamine transporter cDNA probe were detectable in RBL cells but not in JAr choriocarcinoma cells. Similarly, there was no evidence for the expression of the vesicular monoamine transporter as assessed by Northern blot analysis in normal trophoblasts which were maintained in culture to differentiate to form multinucleated syncytial cells. It is concluded that the trophoblasts and choriocarcinoma cells express the plasma membrane serotonin transporter but not the vesicular monoamine transporter.

摘要

我们研究了绒毛膜癌细胞和正常滋养层细胞中质膜5-羟色胺转运体及囊泡单胺转运体的功能表达。将同时表达这两种转运体的大鼠嗜碱性白血病细胞系RBL 2H3细胞用于比较。通过完整细胞中存在的5-羟色胺转运活性评估发现,绒毛膜癌细胞JAr和BeWo拥有质膜5-羟色胺转运体,该转运活性依赖于Na(+),并对三环和非三环抗抑郁药的抑制敏感。通过测量洋地黄皂苷通透细胞中的5-羟色胺转运来确定这些细胞中囊泡单胺转运体的活性。通透细胞中的转运非常缓慢,不受ATP刺激,且对利血平的抑制不敏感。在类似条件下,RBL细胞中可证明有囊泡单胺转运体活性,其受ATP刺激,并可被利血平、巴弗洛霉素A1(一种V型H(+)泵抑制剂)和羰基氰对三氟甲氧基苯腙(一种消散跨膜H+梯度的质子载体)抑制。与这些发现一致,在RBL细胞中可检测到与囊泡单胺转运体cDNA探针杂交的mRNA转录本,但在JAr绒毛膜癌细胞中未检测到。同样,通过Northern印迹分析评估,在培养以分化形成多核合体细胞的正常滋养层细胞中,也没有囊泡单胺转运体表达的证据。结论是,滋养层细胞和绒毛膜癌细胞表达质膜5-羟色胺转运体,但不表达囊泡单胺转运体。

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