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Related Experiment Videos

Multiple cyclic nucleotide phosphodiesterases in human trabecular meshwork cells.

L Zhou1, W J Thompson, D E Potter

  • 1Department of Pharmacology and Toxicology, Morehouse School of Medicine, Atlanta, Georgia 30310, USA.

Investigative Ophthalmology & Visual Science
|July 7, 1999
PubMed
Summary

Human trabecular meshwork cells possess phosphodiesterase 4, 5, and 7 enzymes. These findings suggest that specific phosphodiesterase inhibitors could enhance antiglaucoma drug efficacy by modulating cyclic nucleotide pathways.

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Area of Science:

  • Ophthalmology
  • Molecular Biology
  • Pharmacology

Background:

  • Trabecular meshwork (TM) cells regulate intraocular pressure.
  • Cyclic nucleotides (cAMP, cGMP) play roles in TM cell function.
  • Phosphodiesterases (PDEs) modulate cyclic nucleotide levels by hydrolysis.

Purpose of the Study:

  • To characterize PDE isozyme activities in human and porcine TM cells.
  • To identify specific PDE isoforms present in TM cells.
  • To explore the potential of PDE inhibitors in glaucoma treatment.

Main Methods:

  • Radioimmunoassay to measure cyclic nucleotide levels (cAMP, cGMP).
  • Treatment of human TM cells with PDE isoform-selective inhibitors (e.g., rolipram, E4021).
  • Anion-exchange chromatography to fractionate PDE activities.

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  • Reverse transcription-polymerase chain reaction (RT-PCR) and sequencing to detect PDE transcripts.

    • Main Results:

    • Selective PDE4 and PDE5 inhibitors increased cyclic nucleotide accumulation in human TM cells.
    • PDE activity assays revealed distinct cAMP and cGMP hydrolyzing peaks.
    • Human TM cells expressed mRNA for PDE4, PDE5, and PDE7 isoforms.
    • Porcine TM cells expressed mRNA for PDE4 and PDE5 isoforms.

    Conclusions:

    • Human TM cells express functional PDE4, PDE5, and PDE7 isoforms.
    • PDE inhibitors targeting these isoforms may enhance the efficacy of glaucoma medications.
    • This research provides a basis for developing novel glaucoma therapies targeting PDE pathways.