May 2004
Volume 45, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2004
Immunolocalization of CYP1B1 in Normal, Human Adult and Fetal Eyes and Primary Congenital Glaucoma (PCG) Eyes
Author Affiliations & Notes
  • M. Doshi
    Ophthalmology & Visual Sciences, University of Illinois, Chicago, IL
  • C. Marcus
    College of Pharmacy, University of New Mexico Health Sciences Center, Albuquerque, NM
  • B.A. Bejjani
    Genetics and Pediatrics, Washington State University and Sacred Heart Medical Center, Spokane, WA
  • D.P. Edward
    Ophthalmology & Visual Sciences, University of Illinois, Chicago, IL
  • Footnotes
    Commercial Relationships  M. Doshi, None; C. Marcus, None; B.A. Bejjani, None; D.P. Edward, None.
  • Footnotes
    Support  Gift from Ms. Schild; NEI: R01 EY11780; NIH grant: 1 R01 ES09878
Investigative Ophthalmology & Visual Science May 2004, Vol.45, 4681. doi:
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      M. Doshi, C. Marcus, B.A. Bejjani, D.P. Edward; Immunolocalization of CYP1B1 in Normal, Human Adult and Fetal Eyes and Primary Congenital Glaucoma (PCG) Eyes . Invest. Ophthalmol. Vis. Sci. 2004;45(13):4681.

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

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Abstract

Abstract: : Purpose: CYP1B1 is a cytochrome P450 enzyme implicated in the hereditary form of PCG. CYP1B1 catalyzes the 4–hydroxylation of estrogen and activates various promutagens. In the developing mouse eye, CYP1B1 mRNA is detected in the ciliary body and epithelium and neuroepithelium. This experiment was designed to immunolocalize CYP1B1 protein in the various ocular structures of the normal, human adult and fetal eyes and PCG eyes. Methods: Normal (n=10), fetal (n=9), and enucleated specimens of PCG eyes with end stage disease and mutation status unknown (n=4) were immunolabeled using a polyclonal antibody against CYP1B1 using indirect immunofluorescence and then compared with appropriate controls (ductal breast adenocarcinoma). The intensity of immunolabeling of the various ocular structures was assessed semiquantitatively. Results: In the anterior segment anti–CYP1B1 immunoreactivity (IR) was seen mainly in the ciliary epithelium in fetal eyes with decreased label intensity in adult and PCG eyes. Also, fetal eyes demonstrated moderate and mild IR in the iris epithelium/dilator muscle and corneal keratocytes, respectively which decreased in intensity in adult and PCG eyes. Mild IR was also noted in the fetal corneal epithelium. Anti–CYP1B1 IR was absent in the trabecular endothelium in all eyes. In the posterior segment, mild anti CYP1B1 IR was seen mainly in fetal eyes along the Muller cell processes adjacent to the external limiting membrane and mild IR was noted in the retinal pigment epithelium. Such IR in the posterior segment was absent in adult and PCG eyes. In PCG eyes, the ocular structures that demonstrated anti–CYP1B1 IR showed decreased staining intensity compared to the normal eyes. Conclusion: The immunolocalization of CYP1B1 mainly to the ciliary epithelium in human anterior segment confirms findings previously described in the developing mouse eye. It also suggests that CYP1B1 mutations in PCG might directly or indirectly involve factors produced in the ciliary epithelium that modulate the development of the trabecular meshwork/angle structures resulting in impaired aqueous outflow seen in PCG.

Keywords: ciliary body • microscopy: light/fluorescence/immunohistochemistry • proteins encoded by disease genes 
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