1,25-Dihydroxyvitamin D3-mediated vesicular calcium transport in intestine: dose-response studies.
In order to further test the validity of the vesicular transport model of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3)-stimulated intestinal calcium absorption, dose-response studies were undertaken. Using previously established methodology for subcellular fractionation following 45Ca absorption from in situ ligated duodenal loops, radionuclide levels were found to increase gradually in endocytic vesicles prepared from 1,25(OH)2D3-treated (+D) chicks relative to controls (-D) achieving a plateau at greater than or equal to 260 pmol seco-steroid. By comparison, lysosomal 45Ca levels increased more readily, having +D/-D ratios of 1.88 +/- 0.35, 2.21 +/- 0.05, 2.17 +/- 0.88, 2.31 +/- 0.25, and 2.15 +/- 0.47 after 0.0104, 0.052, 0.26, 1.3, or 6.5 nmol of 1,25(OH)2D3, respectively. Net intestinal calcium absorption, as judged by appearance of 45Ca in the serum for the same range of doses, rose gradually to a plateau value at greater than or equal to 260 pmol. Since lysosomal 45Ca levels were maximally increased at 1,25(OH)2D3 doses lower than those required for fully stimulated transport, it was concluded that lysosomes are still candidates for cellular calcium carriers, but that other elements of the transport pathway are required. Analyses of gradient fractions for calbindin-D28K (the vitamin D-induced calcium binding protein), and potential 1,25(OH)2D3-mediated changes in vesicular ATPase (microtubule motive power for transcellular delivery of calcium) failed to identify the missing components.
Nemere
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Division of Biomedical Sciences, University of California, Riverside 92521.
Norman
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D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D001693:Biological Transport, Active; D002117:Calcitriol / Q000494:pharmacology*; D002118:Calcium / Q000493:pharmacokinetics*; D002645:Chickens; D004305:Dose-Response Relationship, Drug; D004386:Duodenum / Q000187:drug effects / Q000378:metabolism*; D004705:Endocytosis / Q000187:drug effects*; D004721:Endoplasmic Reticulum / Q000378:metabolism; D007408:Intestinal Absorption; D008247:Lysosomes / Q000032:analysis; D008297:Male; D014808:Vitamin D Deficiency / Q000378:metabolism*
D000251:Adenosine Triphosphatases; D002117:Calcitriol; D002118:Calcium