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Related Experiment Videos

Nucleotide receptor-mediated decrease of tight-junctional permeability in cultured human cervical epithelium

G I Gorodeski1, D E Peterson, B J De Santis

  • 1Department of Reproductive Biology, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106, USA.

The American Journal of Physiology
|June 1, 1996
PubMed
Summary

Extracellular ATP acutely alters human cervical cell electrical conductance in two phases. Phase II involves decreased tight junction permeability, confirmed by reduced ion mobility.

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Area of Science:

  • Epithelial Physiology
  • Cell Biology
  • Ion Transport

Background:

  • Extracellular adenosine triphosphate (ATP) is known to modulate transepithelial electrical conductance (GT) in various cell types.
  • ATP elicits a biphasic response in human cervical cells: an initial rapid increase (Phase I) followed by a sustained decrease (Phase II) in GT.
  • The precise mechanisms underlying the Phase II GT decrease, particularly the role of tight junctions, require further elucidation.

Purpose of the Study:

  • To investigate whether the Phase II decrease in transepithelial electrical conductance (GT) induced by extracellular ATP is mediated by reduced tight junction permeability.
  • To analyze the effect of ATP on the relative mobilities of chloride (Cl-) versus sodium (Na+) ions (uCl/uNa) as an indicator of tight junction function.

Main Methods:

Related Experiment Videos

  • Measurement of dilution potential (Vdil) to calculate the relative ion mobility ratio (uCl/uNa) across cultured human cervical cells.
  • Induction of Vdil by altering NaCl concentrations in luminal or subluminal solutions.
  • Assessment of the impact of various stimuli (ATP, hydrostatic/hypertonic gradients, low extracellular Ca2+, ionomycin) on GT and uCl/uNa.

    • Main Results:

    • The baseline uCl/uNa ratio across cervical cell cultures was 1.27, differing from free solution values (1.52).
    • Phase II of the ATP response significantly decreased uCl/uNa to 1.24, an effect abrogated by low extracellular Ca2+.
    • Stimuli that increased GT without affecting tight junctions (hydrostatic/hypertonic gradients, Phase I, ionomycin) did not alter uCl/uNa.

    Conclusions:

    • The Phase II decrease in transepithelial electrical conductance (GT) across cultured human cervical epithelium is primarily mediated by a reduction in tight junction permeability.
    • Extracellular ATP acutely regulates tight junction function in cervical cells, impacting ion selectivity.
    • Calcium ions play a crucial role in maintaining tight junction integrity and mediating the ATP-induced decrease in their permeability.