The alpha(1G)-subunit of a voltage-dependent Ca(2+) channel is localized in rat distal nephron and collecting duct.
The molecular type and localization of calcium channels along the nephron are not well understood. In the present study, we assessed the distribution of the recently identified alpha(1G)-subunit encoding a voltage-dependent calcium channel with T-type characteristics. Using a RNase protection assay, alpha(1G)-mRNA levels in kidney regions were determined as inner medulla >> outer medulla congruent with cortex. RT-PCR analysis of microdissected rat nephron segments revealed alpha(1G) expression in the distal convoluted tubule (DCT), in the connecting tubule and cortical collecting duct (CT+CCD), and inner medullary collecting duct (IMCD). alpha(1G) mRNA was expressed in the IMCD cell line mIMCD-3. Single- and double-labeling immunohistochemistry and confocal laser microscopy on semithin paraffin sections of rat kidneys by using an anti-alpha(1G) antibody demonstrated a distinct labeling at the apical plasma membrane domains of DCT cells, CT principal cells, and IMCD principal cells.
Andreasen
D
D
Department of Physiology and Pharmacology, University of Southern Denmark-Odense University, DK-5000 Odense, Denmark.
Jensen
B L
BL
Hansen
P B
PB
Kwon
T H
TH
Nielsen
S
S
Skøtt
O
O
D000818:Animals; D000918:Antibody Specificity; D020747:Calcium Channels, T-Type / Q000235:genetics / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D003001:Cloning, Molecular; D007124:Immunoenzyme Techniques; D007685:Kidney Tubules, Collecting / Q000378:metabolism* / Q000648:ultrastructure; D008297:Male; D018613:Microscopy, Confocal; D009399:Nephrons / Q000378:metabolism* / Q000648:ultrastructure; D023282:Nuclease Protection Assays; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014018:Tissue Distribution
C500994:Cacna1g protein, rat; D020747:Calcium Channels, T-Type; D012333:RNA, Messenger