Vitamin D receptor mRNA measured in leukocytes with the TaqMan fluorogenic detection system

Effect of calcitriol administration

Laura Soldati, Donatella Adamo, Cristiana Bianchin, Teresa Arcidiacono, Annalisa Terranegra, Maria Luisa Bianchi, Stefano Mora, Daniele Cusi, Giuseppe Vezzoli

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Background: The aim of the present study was to investigate the interactions between the circulating concentrations of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] and the mRNA concentration of its specific nuclear receptor in human leukocytes. Methods: We measured vitamin D receptor (VDR) mRNA extracted from leukocytes by use of TaqMan fluorescence analysis applied to the reverse transcription-PCR (RT-PCR) technique in 16 volunteers before and after calcitriol administration. VDR mRNA was also measured in leukocytes from calcium-stone-formers (37 hypercalciuric and 34 normocalciuric patients). The relationship between VDR mRNA concentrations and genetic VDR polymorphisms was analyzed in these patients. Results: Imprecision (CV) of RT-PCR was 1.3% within assay (n = 10) and 1.7% between assays (n = 4). Oral 1,25(OH)2D3 increased mean (SE) serum 1,25(OH) 2D31.6 (0.3)-fold and VDR mRNA 1.6 (0.1)-fold 8 h after administration. The maximum VDR mRNA was reached 3.6 (1.3) h after 1,25(OH) 2D3 ingestion. No differences in leukocyte VDR mRNA concentrations were found between normocalciuric and hypercalciuric stone-formers in the absence of stimulation. Finally, no association was found between VDR mRNA concentrations and genetic VDR polymorphisms in stone-formers. Conclusions: The TaqMan RT-PCR assay is a rapid and accurate method to measure VDR mRNA, and leukocytes are a useful model to study VDR and 1,25(OH) 2D3 interactions. In humans, VDR mRNA is increased by agonist 1,25(OH)2D3, a finding resembling previously reported results obtained in cellular and animal models.

Original languageEnglish
Pages (from-to)1315-1321
Number of pages7
JournalClinical Chemistry
Volume50
Issue number8
DOIs
Publication statusPublished - Aug 2004

Fingerprint

Calcitriol Receptors
Calcitriol
Leukocytes
Messenger RNA
Transcription
Reverse Transcription
Assays
Polymorphism
Polymerase Chain Reaction
Cytoplasmic and Nuclear Receptors
Volunteers
Animals
Animal Models
Eating
Fluorescence

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this

Vitamin D receptor mRNA measured in leukocytes with the TaqMan fluorogenic detection system : Effect of calcitriol administration. / Soldati, Laura; Adamo, Donatella; Bianchin, Cristiana; Arcidiacono, Teresa; Terranegra, Annalisa; Bianchi, Maria Luisa; Mora, Stefano; Cusi, Daniele; Vezzoli, Giuseppe.

In: Clinical Chemistry, Vol. 50, No. 8, 08.2004, p. 1315-1321.

Research output: Contribution to journalArticle

Soldati, Laura ; Adamo, Donatella ; Bianchin, Cristiana ; Arcidiacono, Teresa ; Terranegra, Annalisa ; Bianchi, Maria Luisa ; Mora, Stefano ; Cusi, Daniele ; Vezzoli, Giuseppe. / Vitamin D receptor mRNA measured in leukocytes with the TaqMan fluorogenic detection system : Effect of calcitriol administration. In: Clinical Chemistry. 2004 ; Vol. 50, No. 8. pp. 1315-1321.
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abstract = "Background: The aim of the present study was to investigate the interactions between the circulating concentrations of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] and the mRNA concentration of its specific nuclear receptor in human leukocytes. Methods: We measured vitamin D receptor (VDR) mRNA extracted from leukocytes by use of TaqMan fluorescence analysis applied to the reverse transcription-PCR (RT-PCR) technique in 16 volunteers before and after calcitriol administration. VDR mRNA was also measured in leukocytes from calcium-stone-formers (37 hypercalciuric and 34 normocalciuric patients). The relationship between VDR mRNA concentrations and genetic VDR polymorphisms was analyzed in these patients. Results: Imprecision (CV) of RT-PCR was 1.3{\%} within assay (n = 10) and 1.7{\%} between assays (n = 4). Oral 1,25(OH)2D3 increased mean (SE) serum 1,25(OH) 2D31.6 (0.3)-fold and VDR mRNA 1.6 (0.1)-fold 8 h after administration. The maximum VDR mRNA was reached 3.6 (1.3) h after 1,25(OH) 2D3 ingestion. No differences in leukocyte VDR mRNA concentrations were found between normocalciuric and hypercalciuric stone-formers in the absence of stimulation. Finally, no association was found between VDR mRNA concentrations and genetic VDR polymorphisms in stone-formers. Conclusions: The TaqMan RT-PCR assay is a rapid and accurate method to measure VDR mRNA, and leukocytes are a useful model to study VDR and 1,25(OH) 2D3 interactions. In humans, VDR mRNA is increased by agonist 1,25(OH)2D3, a finding resembling previously reported results obtained in cellular and animal models.",
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T1 - Vitamin D receptor mRNA measured in leukocytes with the TaqMan fluorogenic detection system

T2 - Effect of calcitriol administration

AU - Soldati, Laura

AU - Adamo, Donatella

AU - Bianchin, Cristiana

AU - Arcidiacono, Teresa

AU - Terranegra, Annalisa

AU - Bianchi, Maria Luisa

AU - Mora, Stefano

AU - Cusi, Daniele

AU - Vezzoli, Giuseppe

PY - 2004/8

Y1 - 2004/8

N2 - Background: The aim of the present study was to investigate the interactions between the circulating concentrations of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] and the mRNA concentration of its specific nuclear receptor in human leukocytes. Methods: We measured vitamin D receptor (VDR) mRNA extracted from leukocytes by use of TaqMan fluorescence analysis applied to the reverse transcription-PCR (RT-PCR) technique in 16 volunteers before and after calcitriol administration. VDR mRNA was also measured in leukocytes from calcium-stone-formers (37 hypercalciuric and 34 normocalciuric patients). The relationship between VDR mRNA concentrations and genetic VDR polymorphisms was analyzed in these patients. Results: Imprecision (CV) of RT-PCR was 1.3% within assay (n = 10) and 1.7% between assays (n = 4). Oral 1,25(OH)2D3 increased mean (SE) serum 1,25(OH) 2D31.6 (0.3)-fold and VDR mRNA 1.6 (0.1)-fold 8 h after administration. The maximum VDR mRNA was reached 3.6 (1.3) h after 1,25(OH) 2D3 ingestion. No differences in leukocyte VDR mRNA concentrations were found between normocalciuric and hypercalciuric stone-formers in the absence of stimulation. Finally, no association was found between VDR mRNA concentrations and genetic VDR polymorphisms in stone-formers. Conclusions: The TaqMan RT-PCR assay is a rapid and accurate method to measure VDR mRNA, and leukocytes are a useful model to study VDR and 1,25(OH) 2D3 interactions. In humans, VDR mRNA is increased by agonist 1,25(OH)2D3, a finding resembling previously reported results obtained in cellular and animal models.

AB - Background: The aim of the present study was to investigate the interactions between the circulating concentrations of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] and the mRNA concentration of its specific nuclear receptor in human leukocytes. Methods: We measured vitamin D receptor (VDR) mRNA extracted from leukocytes by use of TaqMan fluorescence analysis applied to the reverse transcription-PCR (RT-PCR) technique in 16 volunteers before and after calcitriol administration. VDR mRNA was also measured in leukocytes from calcium-stone-formers (37 hypercalciuric and 34 normocalciuric patients). The relationship between VDR mRNA concentrations and genetic VDR polymorphisms was analyzed in these patients. Results: Imprecision (CV) of RT-PCR was 1.3% within assay (n = 10) and 1.7% between assays (n = 4). Oral 1,25(OH)2D3 increased mean (SE) serum 1,25(OH) 2D31.6 (0.3)-fold and VDR mRNA 1.6 (0.1)-fold 8 h after administration. The maximum VDR mRNA was reached 3.6 (1.3) h after 1,25(OH) 2D3 ingestion. No differences in leukocyte VDR mRNA concentrations were found between normocalciuric and hypercalciuric stone-formers in the absence of stimulation. Finally, no association was found between VDR mRNA concentrations and genetic VDR polymorphisms in stone-formers. Conclusions: The TaqMan RT-PCR assay is a rapid and accurate method to measure VDR mRNA, and leukocytes are a useful model to study VDR and 1,25(OH) 2D3 interactions. In humans, VDR mRNA is increased by agonist 1,25(OH)2D3, a finding resembling previously reported results obtained in cellular and animal models.

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