The alpha(1G)-subunit of a voltage-dependent Ca(2+) channel is localized in rat distal nephron and collecting duct

Am J Physiol Renal Physiol. 2000 Dec;279(6):F997-1005. doi: 10.1152/ajprenal.2000.279.6.F997.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibody Specificity
  • Calcium Channels, T-Type / genetics
  • Calcium Channels, T-Type / metabolism
  • Cell Membrane / metabolism
  • Cells, Cultured
  • Cloning, Molecular
  • Immunoenzyme Techniques
  • Kidney Tubules, Collecting / metabolism*
  • Kidney Tubules, Collecting / ultrastructure
  • Male
  • Microscopy, Confocal
  • Nephrons / metabolism*
  • Nephrons / ultrastructure
  • Nuclease Protection Assays
  • RNA, Messenger / biosynthesis
  • Rats
  • Rats, Sprague-Dawley
  • Reverse Transcriptase Polymerase Chain Reaction
  • Tissue Distribution

Substances

  • Cacna1g protein, rat
  • Calcium Channels, T-Type
  • RNA, Messenger