April 2011
Volume 52, Issue 14
ARVO Annual Meeting Abstract  |   April 2011
Ocular Distribution of the Enzyme Prostamide/Prostaglandin F Synthase
Author Affiliations & Notes
  • Lindsey H. Millard
    University of Arizona, Tucson, Arizona
  • William D. Stamer
    University of Arizona, Tucson, Arizona
  • David F. Woodward
    Bioll Sci RD-2C, Allergan, Inc, Irvine, California
  • Kikuko Watanabe
    Division of Life Sciences, University of E. Asia, Shimonoseki, Japan
  • Sanae Hagesawa-Ishii
    Department of Pathology, Institute for Developmental Research, Kasugai, Japan
  • Atsuyoshi Shimada
    Department of Pathology, Institute for Developmental Research, Kasugai, Japan
  • Footnotes
    Commercial Relationships  Lindsey H. Millard, Allergan (R); William D. Stamer, Allergan (F, R); David F. Woodward, Allergan (E); Kikuko Watanabe, Allergan (F, R); Sanae Hagesawa-Ishii, Allergan (R); Atsuyoshi Shimada, Allergan (F, R)
  • Footnotes
    Support  Allergan
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 1514. doi:
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      Lindsey H. Millard, William D. Stamer, David F. Woodward, Kikuko Watanabe, Sanae Hagesawa-Ishii, Atsuyoshi Shimada; Ocular Distribution of the Enzyme Prostamide/Prostaglandin F Synthase. Invest. Ophthalmol. Vis. Sci. 2011;52(14):1514.

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      © ARVO (1962-2015); The Authors (2016-present)

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Purpose: : Prostamide/prostaglandin F synthase (PM/PGFS) is an enzyme belonging to the thioredoxin superfamily. It catalyses the formation of prostamide (PM)F and prostaglandin (PG)F by reducing the precursor endoperoxides PMH2 and PGH2. Since both PMF and PGF lower intraocular pressure, establishing the distribution of this key enzyme in ocular anterior segment tissues represents the first step in determining a role for this enzyme in regulating aqueous humor dynamics.

Methods: : Using a specific rabbit polyclonal antibody (IgG) for PM/PGFS, ocular distribution was determined by Western blotting using ocular tissue homogenates and immunohistochemistry. For immunohistochemical studies, C57BL mice were fixed by systemic perfusion with Bouin’s fixative. Eyes were embedded in paraffin and cut into 6-µm-thick sections. Sections were incubated with the anti-PM/PGFS IgG and then with secondary antibody (EnVision+ System HRP-anti-rabbit [DAKO]). Negative controls involved pre-incubating the anti-PM/PGFS IgG with purified PM/PGFS. Immunoreactions were visualized using diaminobenzidine as chromogen. Subsequently, further preparation with H&E staining was performed to reveal tissue structure and orientation.

Results: : Western blot analysis identified the presence of PM/PGFS in the mouse eye. It had the correct size (21-23 kDa) for the purified enzyme. Immunohistochemical analysis revealed positive staining in the non-pigmented cells of the ciliary processes and the trabecular meshwork. Surprisingly, immunopositive results were obtained in the conjunctiva.

Conclusions: : The presence of PM/PGFS in the trabecular meshwork indicates that the biosynthetic products of PM/PGFS may contribute to the regulation of pressure dependent aqueous humor outflow. A further influence on aqueous humor dynamics may occur at the secretion level, since the enzyme is also present in the ciliary processes.

Keywords: ciliary muscle • trabecular meshwork • enzymes/enzyme inhibitors 

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