Díaz P, Vallejos C, Guerrero I, Riquelme G
Laboratorio de Electrofisiología de Membranas, Fisiología y Biofísica, ICBM, Facultad de Medicina, Universidad de Chile, Santiago, Chile.
Placenta. 2008 Oct;29(10):883-91. doi: 10.1016/j.placenta.2008.07.002. Epub 2008 Aug 15.
The human placental syncytiotrophoblast (hSTB) is a polarized epithelial structure, without paracellular routes, forming the main barrier for materno-fetal exchange. There is ample evidence suggesting the presence of potassium (K(+)) channels in the placental apical membrane; which could contribute to membrane potential and volume regulation. We have therefore examined the K(+) currents of isolated apical membranes from human term placenta using electrophysiological methods: reconstitution of ion channels from apical membranes into giant liposomes (single channel recordings, patch clamp method) or their functional transplantation into Xenopus laevis oocytes (total currents recording, voltage clamp method). Single channel recording experiments show the presence of K(+) channels in the hSTB microvillous membrane sensitive to Tetraethylammonium (TEA) and Barium (Ba(+2)). Patch current activity was diminished 50% and 70% by 20 mmol/L TEA and 5 mmol/L Ba(+2) respectively. The more frequent conductance was approximately 73pS, however several levels of current were detected suggesting the presence of more than one type of K(+) channel. In addition, sodium (Na(+)) sensitivity was detected in the patch current thus, over 10 mmol/L Na(+) reduced the seal current to 38%. These results were corroborated by the total current experiments where the K(+) current elicited in injected oocytes with apical purified membrane was blocked by Ba(+2) and TEA. The total current was also affected by Na(+), becoming larger when a Na(+)-free solution was used. Our results show the existence of at least two types of Ba(+2)-sensitive K(+) channels including a TEA sensitive sub-population, and some of them Na(+) sensitive K(+) channels. These channels could be the conductive pathways proposed previously for this cation in placental hSTB. Our novel contribution has been to successfully obtain K(+) channel recordings in systems suitable for electrophysiological studies of isolated apical membranes.
人胎盘合体滋养层细胞(hSTB)是一种极化上皮结构,没有细胞旁通道,构成母胎交换的主要屏障。有充分证据表明胎盘顶端膜中存在钾(K⁺)通道,这可能有助于膜电位和体积调节。因此,我们采用电生理方法研究了人足月胎盘分离顶端膜的K⁺电流:将顶端膜中的离子通道重构到巨型脂质体中(单通道记录,膜片钳法),或将其功能移植到非洲爪蟾卵母细胞中(总电流记录,电压钳法)。单通道记录实验表明,hSTB微绒毛膜中存在对四乙铵(TEA)和钡(Ba²⁺)敏感的K⁺通道。膜片电流活性分别被20 mmol/L TEA和5 mmol/L Ba²⁺降低了50%和70%。最常见的电导约为73 pS,但检测到几个电流水平,表明存在不止一种类型的K⁺通道。此外,在膜片电流中检测到钠(Na⁺)敏感性,因此,超过10 mmol/L的Na⁺将封接电流降低到38%。这些结果在总电流实验中得到了证实,在用顶端纯化膜注射卵母细胞中引发的K⁺电流被Ba²⁺和TEA阻断。总电流也受Na⁺影响,当使用无Na⁺溶液时电流变大。我们的结果表明至少存在两种对Ba²⁺敏感的K⁺通道,包括一个对TEA敏感的亚群,其中一些是对Na⁺敏感的K⁺通道。这些通道可能是先前提出的该阳离子在胎盘hSTB中的传导途径。我们的新贡献是在适合对分离顶端膜进行电生理研究的系统中成功获得了K⁺通道记录。