Segment-specific expression of sodium-phosphate cotransporters NaPi-IIa and -IIc and interacting proteins in mouse renal proximal tubules
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Sodium-dependent phosphate cotransport in renal proximal tubules (PTs) is heterogeneous with respect to proximal tubular segmentation (S1 vs. S3) and nephron generation (superficial vs. juxtamedullary). In the present study, S1 and S3 segments of superficial and juxtamedullary nephrons were laser-microdissected and mRNA and protein expression of the Na/Pi-cotransporters NaPi-IIa and NaPi-IIc and the PDZ proteins NHERF-1 and PDZK1 determined. Expression of NaPi-IIa mRNA decreased axially in juxtamedullary nephrons. There was no effect of dietary Pi content on NaPi-lla mRNA expression in any proximal tubular segment. The abundance of the NaPi-IIa cotransporter in the brush-border membrane showed inter- and intranephron heterogeneity and increased in response to a low-Pi diet (5 days), suggesting that up-regulation of NaPi-lla occurs via post-transcriptional mechanisms. In contrast, NaPi-IIc mRNA and protein was up-regulated by the low-Pi diet in all nephron generations analysed. NHERF-1 and PDZK1, at both mRNA and protein levels, were distributed evenly along the PTs and did not change after a low-Pi diet.
KeywordsProximal tubule Phosphate reabsorption PDZ protein Laser dissection Immunostaining
Affinity purified anti-NaPi-IIc antibody was kindly provided by Dr. K. Miyamoto and anti-NHERF-1 antibodies were obtained from Dr. E. Weinmann. This work was supported by the Postgraduate Course in Experimental Medicine and Biology of the Medical Faculty of the University of Zürich, Switzerland, by the Swiss National Foundations (Grant 31-65397.01 to H.M.) and the Stiftung für wissenschaftliche Forschung der Universität Zürich.
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