Bo Xuenong, Kim Miran, Nori Stefania L, Schoepfer Ralf, Burnstock Geoffrey, North R Alan
Institute of Molecular Physiology, University of Sheffield, Sheffield, S10 2TN, UK.
Cell Tissue Res. 2003 Aug;313(2):159-65. doi: 10.1007/s00441-003-0758-5. Epub 2003 Jul 4.
A monoclonal antibody was developed to the extracellular domain of the rat P2X4 receptor. The antibody was highly selective among all rat P2X receptor subunits, and recognised only the oligomeric, non-denatured form of the P2X4 receptor. Immunohistochemistry showed an extensive pattern of distribution throughout the central and peripheral nervous systems, the epithelia of ducted glands and airways, smooth muscle of bladder, gastrointestinal tract, uterus, and arteries, uterine endometrium and fat cells. The protein was identified by Western blotting in membrane extracts of these tissues, and the ectodomain antibody immunoprecipitated a protein that was recognised with a P2X4 receptor C terminus antibody. The findings indicate that the P2X4 receptor subunit has a very extensive distribution among mammalian tissues, and this suggests possible new functional roles.
开发了一种针对大鼠P2X4受体细胞外结构域的单克隆抗体。该抗体在所有大鼠P2X受体亚基中具有高度选择性,仅识别P2X4受体的寡聚、非变性形式。免疫组织化学显示其在中枢和外周神经系统、导管腺和气道上皮、膀胱、胃肠道、子宫和动脉的平滑肌、子宫内膜和脂肪细胞中广泛分布。通过蛋白质印迹法在这些组织的膜提取物中鉴定出该蛋白,胞外结构域抗体免疫沉淀出一种可被P2X4受体C末端抗体识别的蛋白。这些发现表明P2X4受体亚基在哺乳动物组织中分布非常广泛,这提示了可能的新功能作用。