Sarangarajan R, Dhabia N, Soleimani M, Baird N, Joiner C
Division of Hematology/Oncology, Department of Pediatrics, Cincinnati Comprehensive Sickle Cell Center, OH 45229, USA.
Biochim Biophys Acta. 1998 Sep 23;1374(1-2):56-62. doi: 10.1016/s0005-2736(98)00130-8.
Erythrocyte sodium hydrogen exchanger (NHE) represents one of a limited number of sodium entry pathway in erythrocytes. At least five NHE isoforms have been identified, differing in tissue specificity, regulatory characteristics, and pharmacological sensitivities. Although physiological characteristics of erythrocyte NHE suggest that the widely expressed NHE-1 isoform may be present, evidence is not conclusive and does not exclude the existence of other isoforms. In this study, Northern blot and reverse transcription-polymerase chain reaction (RT-PCR) analyses were used to test for five NHE isoforms in erythroid cells. Blood from patients with sickle cell disease was depleted of white blood cells (WBC) by passage through leukocyte filters and cellulose column. RT-PCR performed on WBC depleted reticulocyte RNA using a NHE-1 primer set yielded product a of expected size, the sequence of which was identical to the published human NHE-1 sequence. Northern blot analysis of the reticulocyte RNA using a 1.6 kb probe revealed a message of approximately 5.0 kb in size. RT-PCR analysis of rat kidney RNA using primers specific for NHE isoforms -2, -3, -4 and rat brain RNA using primer specific for NHE-5 isoform yielded products of expected size, whereas WBC depleted RNA under identical conditions yielded no products. These results identify the erythroid isoform of the sodium hydrogen exchanger as NHE-1.
红细胞钠氢交换体(NHE)是红细胞中有限的钠进入途径之一。已鉴定出至少五种NHE亚型,它们在组织特异性、调节特性和药理敏感性方面存在差异。尽管红细胞NHE的生理特性表明可能存在广泛表达的NHE-1亚型,但证据并不确凿,也不排除其他亚型的存在。在本研究中,采用Northern印迹法和逆转录-聚合酶链反应(RT-PCR)分析来检测红系细胞中的五种NHE亚型。镰状细胞病患者的血液通过白细胞过滤器和纤维素柱去除白细胞(WBC)。使用NHE-1引物对在去除WBC的网织红细胞RNA上进行RT-PCR,得到预期大小的产物a,其序列与已发表的人类NHE-1序列相同。使用1.6 kb探针对网织红细胞RNA进行Northern印迹分析,显示出大小约为5.0 kb的条带。使用针对NHE亚型-2、-3、-4的特异性引物对大鼠肾脏RNA进行RT-PCR分析,以及使用针对NHE-5亚型的特异性引物对大鼠脑RNA进行RT-PCR分析,均得到预期大小的产物,而在相同条件下,去除WBC的RNA未产生产物。这些结果确定红细胞钠氢交换体的红系亚型为NHE-1。