Hoffman Joseph F, Dodson Alicia, Wickrema Amittha, Dib-Hajj Sulayman D
Department of Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, CT 06520, USA.
Proc Natl Acad Sci U S A. 2004 Aug 17;101(33):12370-4. doi: 10.1073/pnas.0404228101. Epub 2004 Aug 3.
This article concerns the identification of different types of voltage-gated Na(+) channels and of muscarinic and purinergic receptors that are expressed in human erythroid precursor cells and red cell ghosts. We analyzed, by RT-PCR, RNA that was extracted from purified and synchronously growing human erythroid progenitor cells, differentiating from erythroblasts to reticulocytes in 7 to 14 days. These extracts were free of white cell and platelet contamination. Two types of voltage-gated, tetrodotoxin-sensitive Na(+) channels were found. These were Na(v)1.4 and Na(v)1.7, the former known to be present in skeletal muscle and the latter in peripheral nerve. By using a pan Na(+) channel antibody and Western blotting, an immunoreactive channel was detected in ghosts of human red blood cells, consistent with the expression of these two channels. The transcripts for four of the five known subtypes of muscarinic receptors were also identified, including subtypes M2, M3, M4, and M5, whereas subtype M1 was not found. Expression was also detected for the purinergic type receptors P2X(1), P2X(4), P2X(7), and P2Y(1) whereas types P2Y(2), P2Y(4), and P2Y(6) were not found. We also searched for but did not find transcripts for hBNP-1, a type 1b human brain sodium phosphate cotransporter, and cystic fibrosis transmembrane conductance regulator (CFTR). Implications regarding the presence of these different types of channels and receptors in human red blood cells and their functional significance are discussed.
本文涉及在人类红系前体细胞和红细胞影中表达的不同类型电压门控钠通道以及毒蕈碱型和嘌呤能受体的鉴定。我们通过逆转录聚合酶链反应(RT-PCR)分析了从纯化的、同步生长的人类红系祖细胞中提取的RNA,这些细胞在7至14天内从成红细胞分化为网织红细胞。这些提取物无白细胞和血小板污染。发现了两种类型的电压门控、对河豚毒素敏感的钠通道。它们是Na(v)1.4和Na(v)1.7,前者已知存在于骨骼肌中,后者存在于外周神经中。通过使用泛钠通道抗体和蛋白质印迹法,在人类红细胞影中检测到一种免疫反应性通道,这与这两种通道的表达一致。还鉴定出了五种已知毒蕈碱型受体亚型中的四种转录本,包括M2、M3、M4和M5亚型,而未发现M1亚型。还检测到嘌呤能型受体P2X(1)、P2X(4)、P2X(7)和P2Y(1)的表达,而未发现P2Y(2)、P2Y(4)和P2Y(6)型。我们还搜索了1b型人脑钠磷共转运体hBNP-1和囊性纤维化跨膜传导调节因子(CFTR)的转录本,但未找到。讨论了这些不同类型的通道和受体在人类红细胞中的存在及其功能意义。