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Identification of Phosphate Regulated Transcripts in Ileum and Renal Proximal Tubules
Author(s) -
Knöpfel Thomas,
Kampik Nicole,
Hernando Nati,
Rubio Aliaga Isabel,
Biber Jürg,
Wagner Carsten
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.969.12
Subject(s) - ileum , biology , apical membrane , reabsorption , cotransporter , kidney , medicine , pi , paracellular transport , transcriptome , transcellular , homeostasis , endocrinology , microbiology and biotechnology , biochemistry , chemistry , epithelium , gene expression , gene , genetics , organic chemistry , membrane , permeability (electromagnetism) , sodium
Phosphate (Pi) is a vital mineral for every cell in order to maintain energy metabolism, cell signaling and structure. The kidney and intestine are two major organs in maintaining Pi homeostasis by their role in absorbing Pi from the diet and by the reabsorption of filtered Pi. Apical uptake of Pi into intestinal and proximal tubular epithelial cells is well characterized, whereas basolateral efflux mechanisms are poorly understood. The amount of Na‐Pi cotransporters (members of the Slc34 family) expressed at the apical membrane is regulated by changes in dietary Pi intake through mechanisms, which are not fully understood. The aim of this project is to identify transcripts in ileum and renal proximal tubules regulated by dietary Pi. Identified genes may be involved in the transcellular transport of Pi like the unknown basolateral transport pathway or contribute to adaption of apical Pi uptake and metabolism. C57BL/6 male mice (8 weeks old) were fed a high (1.2%) or low (0.1%) Pi diet for 3 days. Epithelial scrapings were prepared from ileum and micro‐dissected renal proximal tubules were collected. Renal and intestinal adaption was confirmed by qPCR for known Pi regulated transcripts. RNA was then analysed using the RNA sequencing service provided by the Functional Genomics Center Zurich. Comparison of the transcriptome of low and high dietary Pi conditions revealed 2029 genes to be differently (at least 2 fold changes) expressed in ileum and 327 in renal proximal tubules. Regulation of selected transcripts was confirmed by qPCR. Pathway analysis is currently being performed and selected candidates for basolateral Pi transporters will be expressed in Xenopus laevis oocytes. Funded by the Swiss National Science Foundation.