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Title: Scintillation autoradiographic localization of 1,25-dihydroxyvitamin D/sub 3/ in chick intestine. [Tritium tracer techniques]

Abstract

The intracellular binding site of 1,25-dihydroxyvitamin D/sub 3/ (1,25(OH)/sub 2/D/sub 3/) was determined via biochemical analysis of radioactive 1,25(OH)/sub 2/D/sub 3/ association with various chick tissues and then by direct autoradiography. When vitamin D-deficient chicks were injected intracardially with doses of tritiated 1,25(OH)/sub 2/D/sub 3/ and killed 2 h later, 2 to 3 times more radioactivity was found in the intestinal mucosa than was present in equal weights of pancreas, parathyroid, or liver tissue. Very little tritium was found in muscle tissue. The intestinally localized radioactivity was predominantly associated with the nuclear chromatin fraction, and binding of 1,25(OH)/sub 2/(/sup 3/H)D/sub 3/ to the nucleus was maximal 2 h after injection and at a dose of at least 0.52 nmol. Using this dose and time period, autoradiographic studies were done on duodenum and thoracic muscle of rachitic chicks injected with radioactive 1,25(OH)/sub 2/D/sub 3/ (11.2 Ci/mol). Thin sections of tissue were prepared for thaw and dry mount scintillation autoradiography as well as simple dip-coating autoradiography. After exposure for 4 to 6 months, a preferential concentration and retention of tritium-labeled 1,25(OH)/sub 2/D/sub 3/ was evident in the nuclei of intestinal villi and in the crypt of Lieberkuehn cells when each of themore » autoradiographic techniques was utilized. Quantitation of the labeled hormone confirms the significant nuclear accumulation in both villi and crypt cells. No such nuclear concentration of silver grains was observed in thoracic muscle cells, and the intestinal localization was abolished when a 100-fold excess of unlabeled 1,25(OH)/sub 2/D/sub 3/ was injected simultaneously with the radioactive hormone. It is concluded that 1,25(OH)/sub 2/D/sub 3/ is bound in a tissue-selective fashion to a high affinity, low capacity site within the nucleus of its intestinal target organ.« less

Authors:
;
Publication Date:
Research Org.:
Univ. of Arizona College of Medicine, Tucson
OSTI Identifier:
6371721
Alternate Identifier(s):
OSTI ID: 6371721
Resource Type:
Journal Article
Journal Name:
Endocrinology; (United States)
Additional Journal Information:
Journal Volume: 104:2
Country of Publication:
United States
Language:
English
Subject:
62 RADIOLOGY AND NUCLEAR MEDICINE; CHOLECALCIFEROL; BIOLOGICAL LOCALIZATION; INTESTINES; SCINTISCANNING; LIVER; MUSCLES; PANCREAS; PARATHYROID GLANDS; TRITIUM; AUTORADIOGRAPHY; CHEMICAL ANALYSIS; CHICKENS; EXPERIMENTAL DATA; GRAPHS; ISOLATED VALUES; TRACER TECHNIQUES; ANIMALS; BETA DECAY RADIOISOTOPES; BETA-MINUS DECAY RADIOISOTOPES; BIRDS; BODY; COUNTING TECHNIQUES; DATA; DATA FORMS; DIAGNOSTIC TECHNIQUES; DIGESTIVE SYSTEM; ENDOCRINE GLANDS; FOWL; GASTROINTESTINAL TRACT; GLANDS; HYDROGEN ISOTOPES; INFORMATION; ISOTOPE APPLICATIONS; ISOTOPES; LIGHT NUCLEI; NUCLEI; NUMERICAL DATA; ODD-EVEN NUCLEI; ORGANS; RADIOISOTOPE SCANNING; RADIOISOTOPES; VERTEBRATES; VITAMIN D; VITAMINS; YEARS LIVING RADIOISOTOPES 550601* -- Medicine-- Unsealed Radionuclides in Diagnostics

Citation Formats

Jones, P.G., and Haussler, M.R. Scintillation autoradiographic localization of 1,25-dihydroxyvitamin D/sub 3/ in chick intestine. [Tritium tracer techniques]. United States: N. p., 1979. Web.
Jones, P.G., & Haussler, M.R. Scintillation autoradiographic localization of 1,25-dihydroxyvitamin D/sub 3/ in chick intestine. [Tritium tracer techniques]. United States.
Jones, P.G., and Haussler, M.R. Thu . "Scintillation autoradiographic localization of 1,25-dihydroxyvitamin D/sub 3/ in chick intestine. [Tritium tracer techniques]". United States.
@article{osti_6371721,
title = {Scintillation autoradiographic localization of 1,25-dihydroxyvitamin D/sub 3/ in chick intestine. [Tritium tracer techniques]},
author = {Jones, P.G. and Haussler, M.R.},
abstractNote = {The intracellular binding site of 1,25-dihydroxyvitamin D/sub 3/ (1,25(OH)/sub 2/D/sub 3/) was determined via biochemical analysis of radioactive 1,25(OH)/sub 2/D/sub 3/ association with various chick tissues and then by direct autoradiography. When vitamin D-deficient chicks were injected intracardially with doses of tritiated 1,25(OH)/sub 2/D/sub 3/ and killed 2 h later, 2 to 3 times more radioactivity was found in the intestinal mucosa than was present in equal weights of pancreas, parathyroid, or liver tissue. Very little tritium was found in muscle tissue. The intestinally localized radioactivity was predominantly associated with the nuclear chromatin fraction, and binding of 1,25(OH)/sub 2/(/sup 3/H)D/sub 3/ to the nucleus was maximal 2 h after injection and at a dose of at least 0.52 nmol. Using this dose and time period, autoradiographic studies were done on duodenum and thoracic muscle of rachitic chicks injected with radioactive 1,25(OH)/sub 2/D/sub 3/ (11.2 Ci/mol). Thin sections of tissue were prepared for thaw and dry mount scintillation autoradiography as well as simple dip-coating autoradiography. After exposure for 4 to 6 months, a preferential concentration and retention of tritium-labeled 1,25(OH)/sub 2/D/sub 3/ was evident in the nuclei of intestinal villi and in the crypt of Lieberkuehn cells when each of the autoradiographic techniques was utilized. Quantitation of the labeled hormone confirms the significant nuclear accumulation in both villi and crypt cells. No such nuclear concentration of silver grains was observed in thoracic muscle cells, and the intestinal localization was abolished when a 100-fold excess of unlabeled 1,25(OH)/sub 2/D/sub 3/ was injected simultaneously with the radioactive hormone. It is concluded that 1,25(OH)/sub 2/D/sub 3/ is bound in a tissue-selective fashion to a high affinity, low capacity site within the nucleus of its intestinal target organ.},
doi = {},
journal = {Endocrinology; (United States)},
number = ,
volume = 104:2,
place = {United States},
year = {1979},
month = {2}
}