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Related Experiment Videos

Thrombin-induced membrane currents in native Xenopus follicles

Q T Nguyen1, J Stinnakre, R Miledi

  • 1Department of Psychobiology, U.C.I. 92715, USA.

Pflugers Archiv : European Journal of Physiology
|January 1, 1996
PubMed
Summary

Thrombin receptors in Xenopus Laevis follicles trigger chloride channel opening via a PKC-regulated pathway. These receptors, located on follicular cells, mediate thrombin-induced currents that desensitize over time.

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Area of Science:

  • Reproductive Biology
  • Cellular Physiology
  • Molecular Pharmacology

Background:

  • Thrombin plays a role in various physiological processes, including oocyte maturation.
  • The specific mechanisms and cellular localization of thrombin receptors in Xenopus Laevis follicles remain incompletely understood.

Purpose of the Study:

  • To investigate the presence and functional properties of thrombin receptors in Xenopus Laevis follicles.
  • To elucidate the signaling pathway involved in thrombin-induced responses in these cells.

Main Methods:

  • Electrophysiological recordings (voltage-clamp) were performed on Xenopus Laevis follicles.
  • Sub-micromolar concentrations of thrombin were applied to induce currents.
  • Protein kinase C (PKC) inhibitors (staurosporine, H7) and hirudin were used to probe the signaling pathway.

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  • Responses were examined under varying osmotic conditions and in defolliculated oocytes.

    • Main Results:

    • Thrombin induced inward chloride currents in Xenopus Laevis follicles voltage-clamped at -60 mV or -80 mV.
    • These currents exhibited desensitization over tens of minutes.
    • Desensitization was blocked by PKC inhibitors.
    • Thrombin responses were inhibited by hirudin, suggesting a specific interaction.
    • Responses were observed in intact follicles but were inconsistent in defolliculated oocytes, indicating the involvement of follicular cells.
    • Thrombin-induced currents were dependent on osmotic conditions and abolished in normal Ringer solution or with sucrose addition.

    Conclusions:

    • Thrombin acts on receptors located on Xenopus Laevis follicular cells.
    • These receptors trigger chloride channel opening via a pathway regulated by Protein Kinase C (PKC).
    • The findings highlight the role of follicular cells in mediating thrombin signaling during oocyte development.