Strom M, Sandgren M E, Brown T A, DeLuca H F
Department of Biochemistry, University of Wisconsin Madison College of Agricultural and Life Sciences 53706.
Proc Natl Acad Sci U S A. 1989 Dec;86(24):9770-3. doi: 10.1073/pnas.86.24.9770.
The level of mRNA encoding the 1,25-dihydroxyvitamin D3 receptor in the intestine of vitamin D-deficient rats given 1,25-dihydroxyvitamin D3 was determined by Northern blot analysis using a 32P-labeled cDNA probe to the 1,25-dihydroxyvitamin D3 receptor. mRNA levels increased 10-fold above deficiency levels at 6 and 12 hr after an intravenous dose of 1,25-dihydroxyvitamin D3, returning to predosing levels at 24 hr. Total receptor protein level determined by an immunoradiometric assay was increased 2-fold at 12 hr. No change in unoccupied receptor levels determined by ligand-binding assay was observed during this period. These results suggest that 1,25-dihydroxyvitamin D3 increases receptor mRNA and total receptor level to maintain constant levels of unoccupied receptor.
使用针对1,25 - 二羟基维生素D3受体的32P标记cDNA探针,通过Northern印迹分析来测定给予1,25 - 二羟基维生素D3的维生素D缺乏大鼠肠道中编码该受体的mRNA水平。静脉注射1,25 - 二羟基维生素D3后6小时和12小时,mRNA水平比缺乏时增加了10倍,24小时时恢复到给药前水平。通过免疫放射分析测定的总受体蛋白水平在12小时时增加了2倍。在此期间,通过配体结合分析测定的未占据受体水平未观察到变化。这些结果表明,1,25 - 二羟基维生素D3增加受体mRNA和总受体水平,以维持未占据受体的恒定水平。