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Related Experiment Videos

No decrease of 1,25(OH)2D3 receptors and duodenal calbindin-D9k in uraemic rats.

A Szabo1, J Merke, M Thomasset

  • 1Department of Internal Medicine, University of Heidelberg, Germany.

European Journal of Clinical Investigation
|October 1, 1991
PubMed
Summary

Uraemia in rats shows increased vitamin D receptors (1,25(OH)2D3) in parathyroids and intestines, despite normal calcium binding protein synthesis. This challenges previous findings on vitamin D resistance in renal failure.

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Area of Science:

  • Endocrinology
  • Nephrology
  • Molecular Biology

Background:

  • Secondary hyperparathyroidism is common in renal failure.
  • Decreased binding of 1,25(OH)2D3 (calcitriol) in uraemia has been linked to this condition.
  • Previous studies used methods that may have affected receptor analysis.

Purpose of the Study:

  • To re-examine 1,25(OH)2D3 binding in uraemic animals using improved chromatin preparation methods.
  • To characterize vitamin D receptors (VDR) in uraemia with enhanced stability.
  • To investigate the regulation of VDR and calcium binding protein (CaBP) synthesis in experimental uraemia.

Main Methods:

  • Chromatin preparations were used for VDR characterization.
  • Protease inhibitors (PMSF, aprotinin) and molybdate were included in extraction buffers.

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  • K-extraction step was omitted to preserve receptor integrity.
  • Nmax, receptor sedimentation constant (S), DNA affinity, KD, and occupied/total receptor ratios were measured.
  • Calbindin-D9k concentration was assessed before and after 1,25(OH)2D3 administration.

    • Main Results:

    • Uraemic animals exhibited significantly higher Nmax for 1,25(OH)2D3 receptors in intestinal mucosa and parathyroids.
    • Receptor sedimentation constant (S), DNA affinity, and KD remained unchanged.
    • The ratio of occupied to total receptors was not significantly altered.
    • Regulation of 1,25(OH)2D3 receptors in response to 1,25(OH)2D3 injection was abnormal.
    • Calbindin-D9k concentration was comparable in uraemic and control rats, indicating unchanged 1,25(OH)2D3-dependent synthesis.

    Conclusions:

    • Experimental uraemia is characterized by an increase in 1,25(OH)2D3 receptors.
    • 1,25(OH)2D3-dependent synthesis of calcium binding protein (CaBP) is not impaired in experimental uraemia.
    • These findings suggest a complex regulation of vitamin D action in uraemia, potentially involving factors beyond receptor number or CaBP synthesis.