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  • Title: Absence of renal 25-hydroxycholecalciferol-1-hydroxylase activity in a pig strain with vitamin D-dependent rickets.
    Author: Winkler I, Schreiner F, Harmeyer J.
    Journal: Calcif Tissue Int; 1986 Feb; 38(2):87-94. PubMed ID: 3082499.
    Abstract:
    Cholecalciferol-1-hydroxylase activities were estimated in renal cortex homogenates and mitochondrial preparations from three groups of 6- to 12-week-old pigs. Five animals suffering from an inherited form of vitamin D-deficiency rickets showed symptoms of florid rickets when used for this study. Six pigs were normocalcemic heterozygous litter mates of the rachitic strain and three pigs were normal controls (German land-race and wild pigs). The renal cortex homogenates and mitochondrial preparations were incubated for 5-30 min at 37 degrees C with 25-(26-27-methyl-3H) OHD3 as substrate. 1,25(OH)2D3 was subsequently identified in normal phase (Zorbax-Sil) and reversed phase (Zorbax-ODS) HPLC eluates. 1-hydroxylase activities were demonstrated in both normal controls and heterozygote offspring and were ten times above minimum detectability of the assay. The km was 255 +/- 74 (SD) and 278 +/- 85 nmol X 1(-1) in controls and heterozygote offspring, respectively. The Vmax in the two groups was between 0.11 and 0.794 and decreased with age of the animals. Km and Vmax did not differ between the two groups. In homozygous, hypocalcemic rachitic animals no 1-hydroxylase activity was detectable in either homogenates or mitochondrial preparations. Addition of kidney homogenate from a rachitic animal to a homogenate from a normal pig did not specifically depress 1-hydroxylase activity in the mixture. Treatment of rachitic pigs with 1.0 microgram/day of 1,25(OH)2D3 for 4 weeks also had no effect on 1-hydroxylase activity. It is concluded that the rachitic pigs suffer from an inborn error of renal 1,25(OH)2D3 production.(ABSTRACT TRUNCATED AT 250 WORDS)
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