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Endothelin receptor B in trabecular meshwork.

Rita Rosenthal1, Lars Choritz, Rüdiger Zorn

  • 1Institut für Klinische Physiologie, Charité, Universitätsmedizin Berlin, Campus Benjamin Franklin, Hindenburgdamm 30, 12200 Berlin, Germany. rita.rosenthal@charite.de

Experimental Eye Research
|August 3, 2007
PubMed
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Endothelin-B receptor (ET-BR) activation causes contraction in bovine trabecular meshwork (TM) and increases intracellular calcium in TM cells. Human TM cells express ET-BR but do not synthesize endothelin-1 (ET-1).

Area of Science:

  • Ophthalmology
  • Molecular Biology
  • Cell Physiology

Background:

  • Endothelin-1 (ET-1), a potent vasoconstrictor, is implicated in primary open-angle glaucoma pathogenesis.
  • ET-1 acts via ET-A receptors (ET-AR) and ET-B receptors (ET-BR), both present in bovine trabecular meshwork (TM).
  • ET-1 primarily induces TM contraction via ET-AR activation.

Purpose of the Study:

  • To investigate the role of ET-BR in TM function.
  • To examine ET-1 synthesis by human TM (HTM) cells.
  • To analyze ET-BR agonist effects on TM contractility and intracellular calcium.

Main Methods:

  • Used a force-length transducer system to assess bovine TM (BTM) contractility with an ET-BR agonist (IRL-1620).
  • Measured intracellular free Ca(2+) ([Ca(2+)](i)) using fura-2AM in cultured BTM and HTM cells.

Related Experiment Videos

  • Investigated ET-AR and ET-BR expression in HTM cells via Western blot and PCR; analyzed prepro-endothelin-1 (ppET-1) and endothelin-1 converting enzyme (ECE-1) expression using RT-PCR.

    • Main Results:

    • IRL-1620 induced BTM contraction (41% of carbachol effect) and transient [Ca(2+)](i) increases in BTM (365%) and HTM (273%) cells.
    • IRL-1620-induced BTM contraction occurred even under calcium-free conditions.
    • ET-AR and ET-BR proteins and mRNAs were detected in HTM cells, but ppET-1 and ECE-1 isoforms were not expressed.

    Conclusions:

    • Functional ET-BR is expressed in both bovine and human TM.
    • Human TM cells express ET-BR but do not appear to synthesize ET-1.
    • Further research is needed to clarify the physiological role of ET-BR in TM and the source of ET-1.