Ahmad I, Korbmacher C, Segal A S, Cheung P, Boulpaep E L, Barnstable C J
Department of Ophthalmology and Visual Science, Yale University School of Medicine, New Haven, CT 06510.
Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10262-6. doi: 10.1073/pnas.89.21.10262.
Apical nonselective cation channels with an average single-channel conductance of 34 +/- 2.3 pS were found in M-1 mouse cortical collecting duct cells. Channel activity is increased by depolarization and abolished by cytoplasmic calcium removal. Cytoplasmic application of 0.1 mM cGMP decreases channel open probability by 27%. cDNAs corresponding to approximately 40% of the coding region of the photoreceptor channel were isolated by the polymerase chain reaction from M-1 cells and a rat kidney cDNA library. The rat kidney-derived sequence differs by a single base, and the M-1-cell-derived sequence differs by only two bases, from the photoreceptor sequence. A second clone from M-1 cells differs by 20 out of 426 bases from the photoreceptor sequence. In all three clones, the deduced amino acid sequence is identical to that of the rat photoreceptor channel. Northern blot analysis of poly(A)+ RNA from M-1 cells reveals the presence of a 3.2-kilobase band hybridizing with a retinal cGMP-gated cation channel probe. The results suggest the expression in M-1 cells of more than one gene coding for nonselective cation channels or channel subunits, one of which is identical to the cGMP-gated cation channel gene of rod photoreceptors.
在M-1小鼠皮质集合管细胞中发现了平均单通道电导为34±2.3 pS的顶端非选择性阳离子通道。通道活性因去极化而增加,因去除细胞质钙而消失。细胞质中加入0.1 mM cGMP可使通道开放概率降低27%。通过聚合酶链反应从M-1细胞和大鼠肾脏cDNA文库中分离出与光感受器通道编码区约40%相对应的cDNA。大鼠肾脏来源的序列与光感受器序列仅相差一个碱基,M-1细胞来源的序列与光感受器序列仅相差两个碱基。来自M-1细胞的第二个克隆与光感受器序列在426个碱基中有20个不同。在所有三个克隆中,推导的氨基酸序列与大鼠光感受器通道的序列相同。对M-1细胞的聚腺苷酸RNA进行的Northern印迹分析显示存在一条与视网膜cGMP门控阳离子通道探针杂交的3.2千碱基条带。结果表明,在M-1细胞中表达了一个以上编码非选择性阳离子通道或通道亚基的基因,其中一个与视杆光感受器的cGMP门控阳离子通道基因相同。