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Differential dependence of regulatory volume decrease behavior in rabbit corneal epithelial cells on MAPK superfamily

Zan Pan1, José E Capó-Aponte, Fan Zhang

  • 1Department of Biological Sciences, State College of Optometry, State University of New York, 33 West 42nd Street, New York, NY 10036, USA.

Experimental Eye Research
|April 3, 2007
PubMed
Summary
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Regulatory volume decrease (RVD) in corneal cells relies on mitogen-activated protein kinase (MAPK) signaling. Swelling activates ERK and SAPK/JNK, preceding ion channel function, while p38 activation follows RVD.

Area of Science:

  • Cell Biology
  • Physiology
  • Molecular Biology

Background:

  • Regulatory Volume Decrease (RVD) is a critical cellular process for maintaining cell volume homeostasis.
  • Mitogen-Activated Protein Kinase (MAPK) pathways are involved in cellular responses to various stimuli, including osmotic stress.
  • Understanding RVD mechanisms in epithelial cells is crucial for corneal health and function.

Purpose of the Study:

  • To investigate the role of MAPK signaling pathways in hypotonicity-induced RVD in rabbit corneal epithelial cells (RCEC).
  • To elucidate the temporal relationship between MAPK activation and ion channel activity during RVD.

Main Methods:

  • RVD was measured in calcein-AM loaded RCEC using a microplate fluorescence reader.
  • Western blot analysis was employed to assess the activation of extracellular signal-regulated kinase (ERK), p38, and stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK).

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  • Specific ion channel inhibitors (Cl- and K+) and MAPK inhibitors were utilized to dissect signaling pathways.

    • Main Results:

    • Hypotonicity induced RVD, restoring cell volume, which was dependent on both Cl- and K+ channel activity.
    • Activation of ERK and SAPK/JNK was time- and tonicity-dependent, preceding ion channel inhibition, and essential for RVD.
    • p38 activation was found to be a consequence of RVD, as its suppression did not inhibit RVD but was blocked when RVD was suppressed.

    Conclusions:

    • Swelling-induced activation of ERK and SAPK/JNK pathways are critical upstream events that precede and facilitate Cl- and K+ channel activation during RVD in RCEC.
    • p38 MAPK activation occurs downstream of, or as a consequence of, the RVD process itself.
    • These findings reveal a distinct temporal and functional interplay between MAPK signaling and ion transport in epithelial cell volume regulation.