Ono S, Kusano E, Muto S, Ando Y, Asano Y
Division of Nephrology, Department of Medicine, Jichi Medical School, 3311-1 Yakushiji, Minamikawachi-machi, Kawachi-gun, Tochigi, 329-04, Japan.
Pflugers Arch. 1997 Nov;434(6):756-63. doi: 10.1007/s004240050462.
The purpose of the present study was to determine whether the renal inner medulla expresses mRNA for the rat epithelial Na+ channel (rENaC) and, if so, to define its regulatory properties using a low-Na+ diet model. We detected alpha, beta and gamma subunit mRNA in rat renal inner medulla using reverse transcriptase-polymerase chain reaction (RT-PCR) with primers specific for rENaC alpha, beta and gamma subunits. Moreover, we have developed a specific probe for the alpha subunit using RT-PCR with rENaC alpha-subunit-specific primers. The resulting cDNA was verified by sequencing and was then used in Northern blot analysis of distal colon, whole kidney and inner medulla. The probe for the rENaC alpha subunit hybridized not only to distal colon RNA but also to inner medulla RNA derived from rats fed a normal diet. Furthermore, we examined the effect of a low-Na+ diet on alpha, beta and gamma subunit mRNA expression of rENaC using full-length cDNA as a probe. A marked elevation of rENaC alpha subunit mRNA abundance in the inner medulla was observed in response to a high plasma aldosterone concentration induced by dietary Na+ deprivation. On the other hand, neither beta nor gamma subunit mRNA expression was enhanced by a low-Na+ diet. From these results, it is suggested that rENaC is responsible for Na+ transport in the renal inner medulla and that is probably regulated via transcriptional control of the alpha subunit of ENaC.
本研究的目的是确定大鼠肾内髓质是否表达大鼠上皮钠通道(rENaC)的mRNA,如果表达,则使用低钠饮食模型确定其调节特性。我们使用针对rENaCα、β和γ亚基的特异性引物,通过逆转录聚合酶链反应(RT-PCR)在大鼠肾内髓质中检测α、β和γ亚基mRNA。此外,我们使用rENaCα亚基特异性引物通过RT-PCR开发了一种针对α亚基的特异性探针。所得cDNA经测序验证,然后用于远端结肠、全肾和内髓质的Northern印迹分析。rENaCα亚基的探针不仅与远端结肠RNA杂交,还与来自正常饮食喂养大鼠的内髓质RNA杂交。此外,我们使用全长cDNA作为探针,研究了低钠饮食对rENaCα、β和γ亚基mRNA表达的影响。在饮食性钠缺乏诱导的高血浆醛固酮浓度作用下,观察到肾内髓质中rENaCα亚基mRNA丰度显著升高。另一方面,低钠饮食并未增强β和γ亚基mRNA的表达。从这些结果表明,rENaC负责肾内髓质中的钠转运,并且可能通过ENaCα亚基的转录控制进行调节。