April 2014
Volume 55, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2014
Metrology of the zonular apparatus of human and non-human primates using fluorescent confocal microscopy: A pilot study
Author Affiliations & Notes
  • Jean-Marie A Parel
    Ophthalmic Biophysics Center, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, FL
    Vision Cooperative Research Centre, Brien Holden Vision Institute, UNSW, Sydney, NSW, Australia
  • Heather Ann Durkee
    Ophthalmic Biophysics Center, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, FL
  • Shawn P Kelly
    Ophthalmic Biophysics Center, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, FL
  • Esdras Arrieta
    Ophthalmic Biophysics Center, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, FL
  • Mariela C Aguilar
    Ophthalmic Biophysics Center, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, FL
  • Yanrong Shi
    Ophthalmology and Visual Science, Washington University School of Medicine in St. Louis, St. Louis, MO
  • Steven Bassnett
    Ophthalmology and Visual Science, Washington University School of Medicine in St. Louis, St. Louis, MO
  • Footnotes
    Commercial Relationships Jean-Marie Parel, None; Heather Durkee, None; Shawn Kelly, None; Esdras Arrieta, None; Mariela Aguilar, None; Yanrong Shi, None; Steven Bassnett, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 749. doi:
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      Jean-Marie A Parel, Heather Ann Durkee, Shawn P Kelly, Esdras Arrieta, Mariela C Aguilar, Yanrong Shi, Steven Bassnett; Metrology of the zonular apparatus of human and non-human primates using fluorescent confocal microscopy: A pilot study. Invest. Ophthalmol. Vis. Sci. 2014;55(13):749.

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

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Abstract
 
Purpose
 

To characterize the zonular apparatus of human and non-human primate (NHP) eyes using indirect immunostaining and laser scanning confocal microscopy (LSCM).

 
Methods
 

The accommodative apparatus of 7 human (55-99 y/o), 9 cynomolgus (4.3-5.8 y/o), 6 baboons (4.0-4.6 y/o) and 1 rhesus (4 y/o) eyes were isolated to expose the lens, zonules, ciliary body, pars plicata, and pars plana. An indirect immunostaining protocol (Bassnett et al, IOVS 2013) was applied using goat-derived MAGP-1 primary antibody to target the microfibril-associated glycoprotein-1 (MAGP-1) molecules of the zonules followed by a fluorophore bound secondary antibody Alexa Fluor 488. DRAQ5 nuclear stain was used to visualize cells within the lens and ciliary processes. A LSCM (TCS SP5, Leica) was used to capture 5, 20, and 63x images that were processed (Advanced Fluorescence Lite, Leica). Individual volumetric images were merged manually (Adobe Photoshop Elements 8.0) to map the zonular pathway.

 
Results
 

The zonule arrangement along with the ciliary processes and lens could be observed in volumetric images revealing a vast network of fine fibers, some originating in the ciliary body and extending on and inserting into the lens capsule at a radial distance that is 5-10% of the total lens diameter. Extra-lenticular and lenticular attachment sites reveal fan-like projections and separation of individual fibers near the lens and ciliary processes. The images show a uniform distribution of zonules around the lens allowing for a circumferentially quasi-continuous transmission of radial forces during accommodation. Volumetric imaging analysis (in process) allows anterior (e.g. 99 y/o human: ~ 435 zonules and 4.5 y/o cynomolgus monkey: ~ 387 zonules) and posterior (e.g. average of two 4.7 y/o cynomolgus monkeys: ~ 418 zonules) zonule density assessment. Improvements are needed to identify the zonular attachment structure at the pars plana as it might affect the accommodation theory.

 
Conclusions
 

The zonular apparatus in human and NHP eyes can be successfully imaged from the lens equator to the pars plana. Zonule metrology (density, lengths, thicknesses) allows cross-species quantification of age-related changes as well as the morphological characterization of zonule transection (Nankivil et al, IOVS 2009) studies.

 
 
Green = zonules; Magenta = lens & ciliary body epithelial cells.
 
Green = zonules; Magenta = lens & ciliary body epithelial cells.
   
Keywords: 404 accommodation • 457 ciliary processes • 472 comparative anatomy  
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