Age-related alterations in calbindin-D28K induction by 1, 25-dihydroxy vitamin D3 in primary cultures of rat renal tubule cells

Ming L. Chen, Monica Boltz, Sylvia Christakos, H. James Armbrecht

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15 Scopus citations


In vivo studies have indicated that renal calbin-din-D28K protein and mRNA levels decrease in adult and old rats, and this decrease parallels the age-associated decline in serum 1, 25-dihydroxyvitamin D3 [1, 25-(OH)2D3] levels. However, diminished renal responsiveness to 1, 25-(OH)2D3 with advancing age could also contribute to decreased calbindin-D28K expression. To study renal responsiveness with age, primary cell cultures were established from the kidney cortices of young (1 month old), adult (10-12 months old), and old (20-24 months old) rats. Cells were incubated in medium K-1 containing 2% fetal calf serum. Calbindin-D28K protein levels were determined by Western blot and enzyme-linked immunosorbent assay. In young animals, the levels of calbindin-D28K declined from 12.1 ± 1.3 μg/mg protein in the intact kidney to 1.6 ± 0.07 μg/mg protein in cells that had been cultured for 3 days in the absence of 1, 25-(OH)2D3. This sharp decline in calbindin-D28K protein concentration moderated by days 6-8. The continuous presence of 10-7 M 1, 25-(OH)2D3 in the medium did not abolish the decline. The low levels of calbindin-D28K in the cells cultured in the absence of 1, 25-(OH)2D3 provided an excellent experimental system in which to compare the response of the cells to 1, 25-(OH)2D3 between age groups. In cultured cells treated with 1, 25-(OH)2D3 for 72 h, calbindin-D28K induction was greater in cells from adult and old animals compared to cells from young animals. The ratios of calbindin-D28K content (with vitamin D/without vitamin D) were 2.2 ± 0.2, 4.7 ± 0.5, and 7.1 ± 1.5 for young, adult, and old cells, respectively. These studies suggested that the observed in vivo decrease in renal calbindin-D28K with age is primarily due to the lowered circulating 1, 25-(OH)2D3.

Original languageEnglish (US)
Pages (from-to)3295-3300
Number of pages6
Issue number6
StatePublished - Jun 1992

All Science Journal Classification (ASJC) codes

  • Endocrinology


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