RT-PCR microlocalization of mRNAs for calbindin D28k and vitamin D receptor in the murine nephron.
The spatial relationship between vitamin D receptor (VDR) and calbindin D28k [calcium binding protein D28k (CaBP-D28k)] gene expression within the murine kidney was studied by localizing their mRNAs in discrete nephron structures using reverse transcription-polymerase chain reaction (RT-PCR). Primers for beta-actin mRNA were used as a control for the presence of tissue during RT-PCR for CaBP-D28k mRNA. mRNA for CaBP-D28k was found only in distal convoluted tubules (DCTs), connecting tubules (CNTs), and cortical collecting ducts (CCDs). In contrast, VDR mRNA was detected in glomeruli, S2 proximal convoluted tubules, cortical thick ascending limbs of Henle's loop, DCTs, CNTs, and initial CCDs. The presence of both VDR and CaBP-D28k mRNA in DCTs, CNTs, and CCDs is consistent with the hypothesis that cacitriol acts via the VDR to stimulate CaBP-D28k synthesis. Conversely, the presence of VDR mRNA in other parts of the nephron suggests that calcitriol has genomically mediated actions within the kidney in addition to stimulation of CaBP-D28k synthesis.
Liu
L
L
Department of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, USA.
Khastgir
A
A
McCauley
J M
JM
Dunn
S T
ST
Morrissey
J H
JH
Christakos
S
S
Hughes
M R
MR
Bourdeau
J E
JE
D000199:Actins / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D064092:Calbindin 1; D064026:Calbindins; D005260:Female; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D015335:Molecular Probes / Q000235:genetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009399:Nephrons / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism*; D018167:Receptors, Calcitriol / Q000235:genetics*; D064030:S100 Calcium Binding Protein G / Q000737:chemistry / Q000235:genetics*; D014018:Tissue Distribution; D014158:Transcription, Genetic
D000199:Actins; C576499:Calb1 protein, mouse; D064092:Calbindin 1; D064026:Calbindins; D015335:Molecular Probes; D012333:RNA, Messenger; D018167:Receptors, Calcitriol; D064030:S100 Calcium Binding Protein G