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

D Andreasen, B L Jensen, P B Hansen, T H Kwon, S Nielsen, O Skøtt

Research output: Contribution to journalJournal articleResearchpeer-review

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.
Original languageEnglish
JournalAmerican Journal of Physiology: Renal Physiology
Volume279
Issue number6
Pages (from-to)F997-1005
ISSN1931-857X
Publication statusPublished - 2000

Keywords

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

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