April 2010
Volume 51, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2010
The Anterior Segment Effects of a Conditional Dicer Knockout in a Rodent Model
Author Affiliations & Notes
  • P. Challa
    Dept of Ophthalmology, Ophthalmology,
    Duke University Eye Center, Durham, North Carolina
  • L. J. Camras
    Dept of Ophthalmology, Ophthalmology,
    Duke University Eye Center, Durham, North Carolina
  • I. D. Navarro
    Dept of Ophthalmology, Ophthalmology,
    Duke Eye Center, Durham, North Carolina
  • C. C. Luna
    Ophthalmology, Department of Ophthalmology,
    Duke University Eye Center, Durham, North Carolina
  • G. Li
    Dept of Ophthalmology, Ophthalmology,
    Duke Eye Center, Durham, North Carolina
  • G. K. Klintworth
    Pathol Ophthal, Duke University Medical Center, Morrisville, North Carolina
  • D. L. Epstein
    Ophthalmology, Department of Ophthalmology,
    Duke Eye Center, Durham, North Carolina
  • P. Gonzalez
    Ophthalmology, Department of Ophthalmology,
    Duke University Eye Center, Durham, North Carolina
  • Footnotes
    Commercial Relationships  P. Challa, None; L.J. Camras, None; I.D. Navarro, None; C.C. Luna, None; G. Li, None; G.K. Klintworth, None; D.L. Epstein, None; P. Gonzalez, None.
  • Footnotes
    Support  NIH Grant K23EY014019; EY016228; EY01894; EY019137; EY05722; Research to Prevent Blindness
Investigative Ophthalmology & Visual Science April 2010, Vol.51, 6431. doi:
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      P. Challa, L. J. Camras, I. D. Navarro, C. C. Luna, G. Li, G. K. Klintworth, D. L. Epstein, P. Gonzalez; The Anterior Segment Effects of a Conditional Dicer Knockout in a Rodent Model. Invest. Ophthalmol. Vis. Sci. 2010;51(13):6431.

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Abstract
 
Purpose:
 

To evaluate the biological relevance of the regulation of gene expression mediated by microRNAs in the cells of the anterior segment of the eye. MicroRNA function was disrupted by constructing a conditional dicer knockout in living mice.

 
Methods:
 

Targeted deletion of Dicer in the anterior chamber of the eye was accomplished by intracameral injection of 5x106 pfu of an adenoviral vector expressing cre recombinase under the CMV promoter in Dicer1tm1Bdh/J mice, which contain loxP sites on either side of exon 23 of the dicer gene. Control mice were injected in parallel with a recombinant adenovirus expressing EGFP under the same promoter and at the same volume and concentration. Alterations in corneal transparency was monitored by direct inspection via bio-microscopy. Morphological alterations in the cornea and outflow pathway were evaluated by microscopic examination of semi-thin sections 2 months after adenoviral injections.

 
Results:
 

Loss of microRNA function by deletion of dicer in the anterior chamber resulted in visible corneal opacification in less than 2 weeks after adenoviral injection. Histological examination of the cornea revealed severe morphological changes in Descemet’s membrane, corneal edema, and loss of corneal epithelial cells (Figure 1). Similarly, the trabecular meshwork showed morphological alterations including a noticeable increase in pigment accumulation.

 
Conclusions:
 

The effects caused by loss of microRNA function through dicer knockdown in the cornea support the concept that regulation of gene expression by microRNAs is important in the maintenance of the normal physiology of adult tissues such as the cornea and the trabecular meshwork.  

 
Keywords: trabecular meshwork • gene modifiers • anterior chamber 
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