Levi M, Lötscher M, Sorribas V, Custer M, Arar M, Kaissling B, Murer H, Biber J
Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas.
Am J Physiol. 1994 Nov;267(5 Pt 2):F900-8. doi: 10.1152/ajprenal.1994.267.5.F900.
Recently, the cDNA for a Na-P(i) cotransport system of rat kidney cortex (NaPi-2) has been identified by expression cloning. Using polyclonal antibodies raised against this renal Na-P(i) cotransport system, and using the polymerase chain reaction after reverse transcription of mRNA in microdissected nephron segments, we recently demonstrated that NaPi-2-related mRNA and protein is expressed in the brush-border membranes (BBM) of the proximal tubules of rat kidney. The purpose of the present study was to study the cellular mechanisms involved in adaptation of rat renal Na-P(i) cotransporter to acute and chronic alterations in dietary P(i). Compared with rats fed chronically (7 days) a high-P(i) diet (1.2%), in rats fed chronically a low-P(i) (0.1%) diet the 3.4-fold increase in BBM Na-P(i) cotransport rate (chronic upregulation) was associated with a 2.2-fold increase in renal cortical NaPi-2 mRNA and a 4.9-fold increase in BBM NaPi-2 protein abundances. In contrast, compared with rats fed chronically (7 day) a high-P(i) diet, in rats fed acutely (2 h) a low-P(i) diet the 1.5-fold increase in Na-P(i) cotransport rate (acute upregulation) was associated with a 1.8-fold increase in NaPi-2 protein but no change in NaPi-2 mRNA abundance. Similarly, compared with rats fed chronically a low-P(i) diet, in rats fed acutely (2 h) a high-P(i) diet the 1.9-fold decrease in Na-P(i) cotransport rate (acute downregulation) was associated with a 3.8-fold decrease in NaPi-2 protein but no change in NaPi-2 mRNA abundance.(ABSTRACT TRUNCATED AT 250 WORDS)
最近,通过表达克隆鉴定出了大鼠肾皮质钠-无机磷共转运系统(NaPi-2)的互补脱氧核糖核酸(cDNA)。我们利用针对该肾钠-无机磷共转运系统产生的多克隆抗体,并在显微切割的肾单位节段中对信使核糖核酸(mRNA)进行逆转录后使用聚合酶链反应,最近证明NaPi-2相关的mRNA和蛋白质在大鼠肾近端小管的刷状缘膜(BBM)中表达。本研究的目的是研究大鼠肾钠-无机磷共转运体适应饮食中无机磷急性和慢性变化所涉及的细胞机制。与长期(7天)喂食高磷(1.2%)饮食的大鼠相比,长期喂食低磷(0.1%)饮食的大鼠,BBM钠-无机磷共转运速率增加3.4倍(慢性上调),同时肾皮质NaPi-2 mRNA增加2.2倍,BBM NaPi-2蛋白质丰度增加4.9倍。相反,与长期(7天)喂食高磷饮食的大鼠相比,急性(2小时)喂食低磷饮食的大鼠,钠-无机磷共转运速率增加1.5倍(急性上调),与NaPi-2蛋白质增加1.8倍相关,但NaPi-2 mRNA丰度无变化。同样,与长期喂食低磷饮食的大鼠相比,急性(2小时)喂食高磷饮食的大鼠,钠-无机磷共转运速率降低1.9倍(急性下调),与NaPi-2蛋白质降低3.8倍相关,但NaPi-2 mRNA丰度无变化。(摘要截短于250字)