CYP27B1 null mice with LacZreporter gene display no 25-hydroxyvitamin D3-1alpha-hydroxylase promoter activity in the skin.
The hormonally active form of vitamin D(3),1alpha,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)], is synthesized in the kidney through a tightly regulated reaction catalyzed by 25-hydroxyvitamin D(3)-1alpha-hydroxylase (1alpha-hydroxylase), the product of the CYP27B1 gene. Through gene targeting in embryonic stem cells, we engineered a mouse strain in which the coding region of the 1alpha-hydroxylase gene is replaced by the genes for beta-galactosidase (lacZ) and neomycin resistance. Null mice produced no detectable 1alpha-hydroxylase transcript. The mice grew normally when maintained on a balanced diet containing 1,25(OH)(2)D(3) but rapidly developed rickets when phosphorus and 1,25(OH)(2)D(3) were restricted. Rickets was curable through administration of 1,25(OH)(2)D(3) but not its biological precursor, 25-hydroxyvitamin D(3). Upon administration of a diet low in calcium and devoid of any form of vitamin D(3), beta-galactosidase activity was detected in the kidneys of the -/- and +/- mice and in placentas harvested from -/- females bred with -/- males. No beta-galactosidase activity was detected in skin sections or in primary keratinocyte cultures from -/- animals. Our results demonstrate we have generated 1alpha-hydroxylase null mice that display phenotypes characteristic of vitamin D-dependency rickets type I. From the histochemical analysis of reporter gene expression in these mice, we conclude that acute 1,25(OH)(2)D(3) deficiency in otherwise healthy animals does not stimulate local production of 1,25(OH)(2)D(3) in the skin. These findings stand in contrast to previously published reports of 1,25(OH)(2)D(3) production in keratinocytes.
Vanhooke
Janeen L
JL
Department of Biochemistry, University of Wisconsin, Madison, WI 53706, USA.
Prahl
Jean M
JM
Kimmel-Jehan
Christine
C
Mendelsohn
Monica
M
Danielson
Eric W
EW
Healy
Kevin D
KD
DeLuca
Hector F
HF
D015090:25-Hydroxyvitamin D3 1-alpha-Hydroxylase / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D001774:Blood Chemical Analysis; D001842:Bone and Bones / Q000033:anatomy & histology / Q000737:chemistry; D017931:DNA Primers; D005260:Female; D018390:Gene Targeting; D017930:Genes, Reporter / Q000235:genetics*; D006651:Histocytochemistry; D015603:Keratinocytes / Q000378:metabolism; D007668:Kidney / Q000378:metabolism; D007763:Lac Operon / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D010920:Placenta / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D012867:Skin / Q000201:enzymology*; D013054:Spectrophotometry, Atomic; D013234:Stem Cells / Q000378:metabolism; D014808:Vitamin D Deficiency / Q000235:genetics* / Q000378:metabolism; D001616:beta-Galactosidase / Q000378:metabolism
D017931:DNA Primers; D015090:25-Hydroxyvitamin D3 1-alpha-Hydroxylase; D001616:beta-Galactosidase