Renal Na+/H+ exchanger isoforms and their regulation by thyroid hormone
1996
Na+ crosses the luminal membrane of the proximal tubule primarily via Na+/H+ exchange (NHE), and NHE activity is influenced by thyroid status. Pharmacological, immunological, and kinetic studies indicate multiple isoforms of NHE, and four full-length cDNAs have been cloned to date. The aims of this study were to determine which NHE mRNAs (NHE1, -2, -3, and -4) were expressed in the rat proximal tubule, the relative abundance of each in the renal cortex, and the effect of thyroid status on their expression. By blot hybridization of poly(A)+ RNA, all NHE isoform mRNAs were detected in the rat renal cortex; NHE1, -2, and -3 in the proximal tubule; and NHE1 and -3 in LLC-PK1 cells. NHE3 mRNA abundance was fourfold higher than the other three isoforms in renal cortex. The effect of thyroid status was assessed in renal cortex from euthyroid, hypothyroid, and hyperthyroid rats. Although none of the NHE mRNA levels was altered in the transition from euthyroid to hypothyroid states, both NHE2 and NHE3 mRNA levels increased 1.5-fold in the transition from hypo- to hyperthyroidism. NHE3 protein, measured by immunoblot with the use of an NHE3-specific antibody, was detected at 83-85 kDa in renal cortex and codistributed on sorbitol gradients with the brush-border marker alkaline phosphatase. No significant difference in NHE3 protein abundance was detected between hypothyroid and hyperthyroid rats. In conclusion, in the renal cortex, the NHE3 isoform predominates at the mRNA level, is expressed in apical membranes, and increases at the mRNA but not the protein levels in response to thyroid hormone treatment, suggesting parallel changes in synthesis and turnover of NHE3 by thyroid hormone.
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