Presence and localization of two vitamin D-dependent calcium binding proteins in kidneys of higher vertebrates.

We looked for the presence of vitamin D-dependent intestinal calcium binding protein (CaBP), relative molecular mass (mol wt) about 10,000, in kidneys of mammals, birds, and reptiles, and compared the localization of this 10,000 mol wt CaBP with the localization of the 28,000 mol wt vitamin D-dependent CaBP. Antiserum directed against rat intestinal CaBP (RICaBP) was used with the unlabeled antibody peroxidase-antiperoxidase technique to localize the 10,000 mol wt CaBP. Kidneys of adult Swiss and DBA/2J mice were positive for the 10,000 mol wt CaBP, whereas kidneys of adult rat, chicken, and two species of lizard were negative. Extracts of adult rat kidney showed no detectable immunoreactivity with the antiserum to RICaBP by radial immunodiffusion assay. However, kidneys of postnatal rats (12 days old) exhibited limited immunocytochemical reactivity and relatively low immunoreactivity (about 1 micrograms/mg protein) for 10,000 mol wt CaBP. In both strains of mice, 10,000 mol wt CaBP was localized predominantly to the distal convoluted tubules, connecting tubules, and collecting tubules of the cortical labyrinth. In 12-day-old rats 10,000 mol wt CaBP was localized to the distal convoluted tubules and cortical ascending thick limbs. Absorption of the antiserum with electrophoretically pure RICaBP eliminated specific reactivity. By dual color staining of mouse kidneys, the population of cells reactive for the 10,000 mol wt CaBP was found to be similar, but not identical, to that population of cells reactive for the 28,000 mol wt CaBP. Absorption of the antiserum to RICaBP with either rat renal CaBP or chicken intestinal CaBP (both 28,000 mol wt CaBPs) had no affect on the positive reactivity of the distal mouse nephron for RICaBP. Conversely, absorption of the antiserum to rat renal CaBP with the 10,000 mol wt RICaBP had no affect on reactivity for the 28,000 mol wt CaBP. Thus, two immunologically distinct CaBPs, mol wt 10,000 and 28,000, are present in the adult mouse nephron; whereas, kidneys of adult rats, chickens, and saurian reptiles apparently contain significant levels of only 28,000 mol wt CaBP. Since the 12-day-old rat nephron contains both the 28,000 mol wt and the 10,000 mol wt CaBP it appears that in the kidney, ontogenetically, and perhaps evolutionarily as well, the 28,000 mol wt CaBP is more highly conserved.