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Related Experiment Videos

CFTR expression in cortical collecting duct cells

K M Todd-Turla1, E Rusvai, A Náray-Fejes-Tóth

  • 1Department of Physiology, Dartmouth Medical School, Lebanon, New Hampshire 03756, USA.

The American Journal of Physiology
|January 1, 1996
PubMed
Summary
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The cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel mRNA is present in mouse and rabbit kidney collecting ducts. CFTR is most abundant in beta-intercalated cells, suggesting a primary role in their function.

Area of Science:

  • Nephrology
  • Molecular Biology
  • Cell Biology

Background:

  • The cystic fibrosis transmembrane conductance regulator (CFTR) is an adenosine 3',5'-cyclic monophosphate-activated chloride channel.
  • CFTR is found in the apical membrane of various epithelial cells and may have a significant role in kidney function.
  • Previous research suggests CFTR expression in the cortical collecting duct (CCD).

Purpose of the Study:

  • To detect and characterize CFTR mRNA expression in the M-1 mouse CCD cell line and immunoselected rabbit CCD cells.
  • To determine the physiological relevance and cellular distribution of CFTR mRNA within the CCD.

Main Methods:

  • Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect CFTR mRNA.
  • Primers were designed to amplify the cDNA sequence of the first nucleotide-binding domain of CFTR.

Related Experiment Videos

  • Northern analysis and sequencing were employed for product identification and characterization.

    • Main Results:

    • CFTR mRNA was detected in both M-1 mouse CCD cells and rabbit CCD cells.
    • Sequencing confirmed 97% homology between rabbit CCD CFTR and human CFTR.
    • Northern analysis revealed a 6.5 kb CFTR mRNA species.
    • CFTR mRNA was found in principal cells, beta-intercalated cells, and alpha-intercalated cells, with significantly higher levels in beta-intercalated cells.

    Conclusions:

    • CFTR mRNA is present in the mouse and rabbit cortical collecting duct.
    • The abundance of CFTR mRNA in beta-intercalated cells suggests a primary role in their function within the CCD.