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1 Renal Division, Vanderbilt University Medical Center, Nashville, Tennessee 37232-2372; and 2 Endocrine-Hypertension Unit, Department of Medicine, Brigham & Women's Hospital Harvard Medical School, Boston, Massachusetts 02115
We previously identified transcripts encoding a
G protein-coupled, extracellular calcium/polyvalent cation-sensing
receptor, RaKCaR, in rat kidney (D. Riccardi, J. Park, W.-S. Lee, G. Gamba, E. M. Brown, and S. C. Hebert. Proc. Natl.
Acad. Sci. USA 92: 131-135, 1994), which was
proposed to provide the mechanism for modulating a variety of renal
functions in response to changes in extracellular
Ca2+ (E. M. Brown. In:
Handbook of Physiology. Bethesda, MD:
Am. Physiol. Soc., 1992, sect. 8, vol. 2, chapt. 39, p. 1841-1916;
and S. C. Hebert. Kidney Int. 50:
2129-2139, 1996). Here, we examine the cellular and regional
distribution of receptor protein by immunofluorescence microscopy using
a polyclonal antibody raised against a 22 amino acid region of the
NH2 terminus of the receptor. The
most intense fluorescence was seen at the basolateral border of
cortical thick ascending limb cells. Basolateral staining for the
receptor was also detected in medullary thick ascending limbs, in
macula densa cells identified by costaining with antibody to brain
nitric oxide synthase, NOS-B1, and in distal convoluted tubule cells
distinguished by costaining for the apical thiazide-sensitive
Na+-Cl
cotransporter. Apical anti-RaKCaR staining was detected at the base of
the brush border of proximal tubules with decreasing intensity from S1
to S3 segments. In cortical collecting ducts, anti-RaKCaR staining was
detected in some, but not all, type A intercalated cells identified by
costaining with anti-H+-ATPase and
anti-AE1
Cl/
exchanger antibodies. The present study demonstrates that RaKCaR
protein is expressed in many different nephron segments and that the
polarity of receptor expression varies with cell type along the
nephron. These results suggest potential roles for the extracellular
Ca2+/polyvalent cation-sensing
receptor in responding to both circulating and urinary concentrations
of divalent minerals and potentially other polyvalent cations (e.g.,
aminoglycoside antibiotics) to modulate nephron function.
calcium-sensing receptor; G protein-coupled receptor; immunofluorescence; macula densa; nitric oxide synthase; AE1 anion exchanger; chloride/bicarbonate exchanger; proton-adenosinetriphosphatase; thiazide-sensitive sodium-chloride cotransporter; nitric oxide synthase; kidney tubules
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