The intracellular distribution of [1-3H]cholecalciferol in the intestine of vitamin D-deficient and -supplemented rats

Wilson, Peter W.F.; Lawson, David; Kodíček, E.

doi:10.1042/bj1030165

Cited by 16 publications

(7 citation statements)

References 15 publications

(6 reference statements)

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“…We have also found (Wilson, Lawson & Kodicek, 1967) that a fraction of the intestinal cell containing nuclei and brush borders had the highest concentration of radioactivity after a large dose of [loc-3H]cholecalciferol. In the present study we were unable to find radioactivity localized in intestinal nuclei after a dose of 0 25,ug.…”

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confidence: 52%

“…Tissue fractionation. The mitochondrial, microsomal and cytoplasmic fractions were obtained by a modified method of Schneider & Hogeboom (1950) as previously described (Wilson et al 1967). The nuclear-debris fraction and the pure nuclei were prepared as described in the preceding paper under method (b) (Lawson et al 1969a).…”

mentioning

confidence: 99%

“…, with the counting solution described previously (Wilson et al 1967). Quenching was corrected for by using the automatic external standardization and correlation curves for 14C and 3H, either individually or combined.…”

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confidence: 99%

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Metabolism of vitamin D. A new cholecalciferol metabolite, involving loss of hydrogen at C-1, in chick intestinal nuclei

Lawson

Wilson

Kodíček

1969

Biochemical Journal

177

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1. A comparison was made of the nature and intestinal intracellular distribution of the metabolites formed in vitamin D-deficient chicks from [4-(14)C]cholecalciferol and [1-(3)H]cholecalciferol. 2. The simultaneous administration of the two radioactive substances showed the presence in blood, liver, intestine, kidney and bone of cholecalciferol, its ester, 25-hydroxycholecalciferol and a further metabolite of cholecalciferol more polar than 25-hydroxycholecalciferol. The (3)H/(14)C ratios in these four radioactive components were the same as that of the dosed material (4.7:1) with the exception of the most polar material. The (3)H/(14)C ratio was lower in the fourth, most polar, metabolite (0.4:1-1.8:1) in all tissues examined, with the exception of blood. 3. In the chick intestine the polar metabolite accounted for almost 70% of the radioactivity in this tissue after a dose of 0.5mug. of [4-(14)C,1-(3)H]cholecalciferol. This polar metabolite from the intestine also had the lowest (3)H/(14)C ratio of all the tissues. It appears that in the chick intestine the polar metabolite reaches a maximum concentration of 1ng./g. of tissue, above which it cannot be increased irrespective of the dose of the vitamin. 4. The intestinal intracellular organelle with the highest concentration of (14)C radioactivity is the nucleus, and this radioactivity is almost entirely due to the polar metabolite with the lowered (3)H/(14)C ratio, in this case <0.2:1. It appears to be further localized in the chromatin of the nuclei. However, about half of the polar metabolite in the intestine is extranuclear. 5. Double-labelled 25-hydroxycholecalciferol was prepared and after its administration to vitamin D-deficient chicks the polar metabolite with the lowered (3)H/(14)C ratio was detected in liver, kidney, intestine, bone, muscle and heart. 6. None of the polar metabolite with the lowered (3)H/(14)C ratio was detected 16hr. after dosing with either the double-labelled vitamin or the double-labelled 25-hydroxycholecalciferol in blood and adipose tissue of vitamin D-deficient chicks, nor in the intestine, liver and kidney of supplemented birds. 7. The reasons for this loss of (3)H relative to (14)C are discussed in relation to possible chemical structures of this new polar metabolite.

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Metabolism of vitamin D. A new cholecalciferol metabolite, involving loss of hydrogen at C-1, in chick intestinal nuclei

Lawson

Wilson

Kodíček

1969

Biochemical Journal

177

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“…Tissue fractionations. The mitochondrial, microsomal and cytoplasmic fractions were obtained by a modified method of Schneider & Hogeboom (1950) as described by Wilson, Lawson & Kodicek (1967). The nuclear debris fractions and the pure nuclei were prepared as before (Lawson et al 1969b (1956).…”

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confidence: 99%

Synthesis of [1,2-3H2]cholecalciferol and metabolism of [4-14C,1,2-3H2]- and [4-14C,1-3H]-cholecalciferol in rachitic rats and chicks

Lawson¹,

Pelc²,

Bell³

et al. 1971

Biochemical Journal

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[1,2-3H2]Cholecalciferol has been synthesized with a specific radioactivity of 508 mCi/mmol by using tristriphenylphosphinerhodium chloride, the homogeneous hydrogen catalyst. With doses of 125ng (5i.u.) of [4-14C,i-3H2]cholecalciferol the tissue distribution in rachitic rats ofcholecalciferol and its metabolites (25-hydroxycholecalciferol and peak P material) was similar to that found in chicken with 500ng doses of the double-labelled vitamin. The only exceptions were rat kidney, with a very high concentration of vitamin D, and rat blood, with a higher proportion of peak P material, containing a substance formed from vitamin D with the loss of hydrogen from C-1. Substance P formed from [4-14C,1,2-3H2]cholecalciferol retained 36% of 3H, the amount expected from its distribution between C-I and C-2, the 3H at C-1 being lost. 25-Hydroxycholecalciferol does not seem to have any specific intracellular localization within the intestine of rachitic chicks. The 3H-deficient substance P was present in the intestine and bone 1 h after a dose of vitamin D and 30min after 25-hydroxycholecalciferol. There was very little 25-hydroxycholecalciferol in intestine at any time-interval, but bone and blood continued to take it up over the 8h experimental period. It is suggested that the intestinal 3H-deficient substance P originates from outside this tissue. The polar metabolite found in blood and which has retained its 3H at C-1 is not a precursor of the intestinal 3H-deficient substance P.

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“…An aliquot o f this material was evaporated with nitrogen in a scintillation vial. The scintillation fluid [13] was added to each sample vial, and radioactivity was counted with a Model 3375 Packard Tri-Carb liquid scintillation counter.…”

Section: Malcriáis and Methodsmentioning

confidence: 99%

Lipid Absorption in Newborn Young of Guinea Pigs Fed a Protein-Deficient Diet during Gestation

Zeman

Widdowson²

1974

Neonatology

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Absorption of ¹⁴C-oleic acid and ¹⁴C-triolein was measured in newborn young of guinea pigs which had been adequately nourished or deprived of protein in the last half of pregnancy. The young of protein-deprived dams absorbed significantly greater amounts of both substances on the basis of amount per gram of intestinal tissue per minute or per gram of body weight per minute. In animals in the deficient group, villi of the jejunum were longer, narrower, and more numerous than in the controls, providing a morphological basis for some of the observed increase.

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The intracellular distribution of [1-3H]cholecalciferol in the intestine of vitamin D-deficient and -supplemented rats

Cited by 16 publications

References 15 publications

Metabolism of vitamin D. A new cholecalciferol metabolite, involving loss of hydrogen at C-1, in chick intestinal nuclei

Metabolism of vitamin D. A new cholecalciferol metabolite, involving loss of hydrogen at C-1, in chick intestinal nuclei

Synthesis of [1,2-3H2]cholecalciferol and metabolism of [4-14C,1,2-3H2]- and [4-14C,1-3H]-cholecalciferol in rachitic rats and chicks

Lipid Absorption in Newborn Young of Guinea Pigs Fed a Protein-Deficient Diet during Gestation

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