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首页> 外文期刊>American Journal of Physiology >A P2X7 receptor stimulates plasma membrane trafficking in the FRTL rat thyrocyte cell line.
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A P2X7 receptor stimulates plasma membrane trafficking in the FRTL rat thyrocyte cell line.

机译：P2X7受体刺激FRTL大鼠甲状腺细胞系中的质膜运输。

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Thyroid cells express a variety of P2Y and P2X purinergic receptor subtypes. G protein-coupled P2Y receptors influence a wide variety of thyrocyte-specific functions; however, functional P2X receptor-gated channels have not been observed. In this study, we used whole cell patch-clamp recording and fluorescence imaging of the plasma membrane marker FM1-43 to examine the effects of extracellular ATP on membrane permeability and trafficking in the Fisher rat thyroid cell line FRTL. We found a cation-selective current that was gated by ATP and 2',3'-O-(4-benzoylbenzoyl)-ATP but not by UTP. The ATP-evoked currents were inhibited by pyridoxal phosphate 6-azophenyl-2',4'-disulfonic acid, adenosine 5'-triphosphate-2',3'-dialdehyde, 100 microM Zn(2+), and 50 microM Cu(2+). Fluorescence imaging revealed pronounced, temperature-sensitive stimulation of exocytosis and membrane internalization by ATP with the same pharmacological profile as observed for activation of current. The EC(50) for ATP stimulation of internalization was 440 microM in saline containing 2 mM Ca(2+) and 2 mM Mg(2+), and 33 microM in low-Mg(2+), nominally Ca(2+)-free saline. Overall, the results are most consistent with activation of a P2X(7) receptor by ATP(4-). However, low permeability to N-methyl-d-glucamine(+) and the propidium cation YO-PRO-1 indicates absence of the cytolytic pore that often accompanies P2X(7) receptor activation. ATP stimulation of internalization occurs in Na(+)-free, Ca(2+)-free, or low-Mg(2+) saline and therefore does not depend on cation influx through the ATP-gated channel. We conclude that ATP activation of a P2X(7) receptor stimulates membrane internalization in FRTL cells via a transduction pathway that does not depend on cation influx.
机译：甲状腺细胞表达多种P2Y和P2X嘌呤能受体亚型。 G蛋白偶联的P2Y受体影响多种甲状腺细胞特异性功能。但是，尚未观察到功能性P2X受体门控通道。在这项研究中，我们使用全细胞膜片钳记录和质膜标记FM1-43的荧光成像来检查细胞外ATP对Fisher大鼠甲状腺细胞系FRTL的膜通透性和运输的影响。我们发现阳离子选择性电流受到ATP和2'，3'-O-（4-苯甲酰基苯甲酰基）-ATP的控制，而不受UTP的控制。 ATP诱发的电流受到吡ido醛磷酸6-偶氮苯基-2'，4'-二磺酸，腺苷5'-三磷酸-2'，3'-二醛，100 microM Zn（2+）和50 microM Cu（ 2+）。荧光成像显示ATP对胞吐作用和膜内在化具有明显的温度敏感性刺激，并且具有与电流激活相同的药理作用。 EC（50）用于ATP刺激的内在化是440 microM在包含2 mM Ca（2+）和2 mM Mg（2+）的盐水中，在33 microM在低Mg（2+），名义上是Ca（2+）不含盐水。总体而言，结果与ATP（4-）激活P2X（7）受体最一致。但是，对N-甲基-d-葡萄糖胺（+）和丙鎓阳离子YO-PRO-1的低渗透性表明缺少通常伴随P2X（7）受体激活的溶细胞孔。 ATP刺激的内化作用发生在无Na（+），无Ca（2+）或低Mg（2+）盐水中，因此不依赖于通过ATP门控通道的阳离子流入。我们得出的结论是，P2X（7）受体的ATP激活通过不依赖于阳离子流入的转导途径刺激FRTL细胞的膜内在化。

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