April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
The Marine Mammal Tear Film Has Unique Attributes
Author Affiliations & Notes
  • Robin Kelleher Davis
    Schepens Eye Res Inst/Harvard Univ, Boston, Massachusetts
  • Nadja Knop
    Univ Eye Clinic Res Lab, University Charite Berlin, Berlin, Germany
  • Erich Knop
    Univ Eye Clinic Res Lab, University Charite Berlin, Berlin, Germany
  • David A. Sullivan
    Schepens Eye Res Inst/Harvard Univ, Boston, Massachusetts
  • Pablo Argueso
    Schepens Eye Res Inst/Harvard Univ, Boston, Massachusetts
  • Footnotes
    Commercial Relationships  Robin Kelleher Davis, None; Nadja Knop, None; Erich Knop, None; David A. Sullivan, None; Pablo Argueso, None
  • Footnotes
    Support  NIH EY05612, NIH EY014847, Arey's Pond Boat Yard, Inc.; Aquarium of Niagara (NY) and Dolphin Quest Oahu (HI) for contibutions of marine mammal tears; Marine Mammal Center (Sausalito, CA) for tissue
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 3723. doi:https://doi.org/
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    • Get Citation

      Robin Kelleher Davis, Nadja Knop, Erich Knop, David A. Sullivan, Pablo Argueso; The Marine Mammal Tear Film Has Unique Attributes. Invest. Ophthalmol. Vis. Sci. 2011;52(14):3723. doi: https://doi.org/.

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

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Abstract

Purpose: : In a previous study we found that pinnipeds lack the lipid layer of the tear film, leading us to hypothesize that mucin-type glycoproteins are the major protective components in the ocular surface of marine mammals. The purpose of this study was to characterize mucin-type O-glycans in marine mammal tears, and to determine whether pinnipeds have meibomian glands.

Methods: : Protein and carbohydrate concentrations of tears collected from humans and marine mammals (with IRB & ACUC approvals) were determined using bicinchoninic acid and sulfuric methods respectively. Samples normalized for protein were electrophoresed on 1% SDS-agarose gels, transferred to nitrocellulose membranes by vacuum blotting, and probed for the T-antigen carbohydrate using Arachis hypogea agglutinin (PNA), followed by chemiluminescence detection. Normal-phase high pressure liquid chromatography (HPLC) was used to analyze O-glycans released by ammonia-based beta-elimination. For histology, ocular tissues from a sea lion necropsy were fixed in formalin, examined using a stereo magnifier, embedded in paraffin, sectioned to 10-20 um thickness using a rotary microtome, and stained.

Results: : Biochemical analysis revealed PNA binding to proteins of high molecular weight, consistent with mucins, in human, sea lion, and dolphin tear samples. The size distribution of PNA-positive bands was different for each species analyzed, with dolphin migrating at the highest molecular weight. Separation of O-glycans by HPLC revealed differences in chromatograms of tears from humans, sea lions, and dolphins. In stereo magnifier analysis, a distinct tarsus was missing. The lid consisted mainly of muscular tissue and underneath the epidermis, there was a whitish granular layer consisting of hair follicles and sebaceous glands. Bundles of holocrine sebaceous acini were arranged between hair follicles. Towards the lid margin, the number of hair follicles decreased and the relative volume of sebaceous glands increased, forming several solitary gland bodies at the margin. The acini drained via ductules into a straight duct with a multi-layered stratified squamous epithelium, which opened onto the outer lid skin.

Conclusions: : Our results indicate that as in humans, mucin-type O-glycans are present in marine mammal tears, with differences in size distribution and O-glycan profiles across species. Pinnipeds have sebaceous glands at the eyelid margin, resembling human meibomian glands in structure, but different in orientation and size.

Keywords: cornea: tears/tear film/dry eye • cornea: surface mucins • glycoconjugates/glycoproteins 
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