April 2014
Volume 55, Issue 13
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ARVO Annual Meeting Abstract  |   April 2014
Age-related changes in composition and geographic distribution of phospholipids in zebrafish lens.
Stephen Barnes; Miranda Collier; Stephen Watts; David E Graves; Janusz H Kabarowski
Author Affiliations & Notes
  • Stephen Barnes
    Pharmacology & Toxicology, Univ of Alabama at Birmingham, Birmingham, AL
  • Miranda Collier
    Department of Chemistry, University of Alabama at Birmingham, Birmingham, AL
  • Stephen Watts
    Department of Biology, University of Alabama at Birmingham, Birmingham, AL
  • David E Graves
    Department of Chemistry, University of Alabama at Birmingham, Birmingham, AL
  • Janusz H Kabarowski
    Department of Microbiology, University of Alabama at Birmingham, Birmingham, AL
  • Footnotes
    Commercial Relationships Stephen Barnes, None; Miranda Collier, None; Stephen Watts, None; David Graves, None; Janusz Kabarowski, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 3572. doi:
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      Stephen Barnes, Miranda Collier, Stephen Watts, David E Graves, Janusz H Kabarowski; Age-related changes in composition and geographic distribution of phospholipids in zebrafish lens.. Invest. Ophthalmol. Vis. Sci. 2014;55(13):3572.

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Abstract
 
Purpose
 

Cell differentiation in the lens may lead to slowed turnover of phospholipids (PL). We have used a zebrafish model of lens development to investigate the change in PL composition of the lens with aging and to determine PL localization within the lens and eye.

 
Methods
 

Zebrafish were euthanized at 2 and 12 months of age. Lens were homogenized and the lipids extracted by the Bligh-Dyer method. The extract was infused into an AB Sciex 5600 TripleTOF mass spectrometer to collect MSMS data at successive unit masses on ions over the range from 200-1200 m/z (mass-to-charge ratio) in the negative and positive modes. LipidView™ was used to deconvolute the data. Eyes were also collected and embedded in 10% gelatin to obtain 16 μm frozen sections that were transferred to conductive glass slides. Matrix (positive spectra, diaminonaphthalene; negative spectra, dihydroxybenzoic acid) was applied to the section by vacuum sublimation. Lipids in the coated section were visualized using imaging mass spectrometry (IMS) with a Bruker Daltoniks Autoflex TOF/TOF. Distributions of PLs were obtained using Fleximaging™ software.

 
Results
 

The principal PLs in the lens in the positive mode were phosphatidylcholines (PC) and sphingomyelins. Each was significantly (p <0.001) increased (per mg tissue) in 12-month old compared to 2-month old fish). In the negative mode, only PC and phosphatidylinositols (PIs) were significantly increased (p <0.001 and p<0.003, respectively) by aging. IMS of putative PL ions (m/z 718.69, 764.51 and 792.75) revealed that they were confined to a thin layer in the outer cortex of the lens, with none in the inner cortex or nuclear region.

 
Conclusions
 

This study has revealed that PCs and PIs are significantly increased by aging in zebrafish lens. Furthermore, IMS data from frozen sections of the lens have demonstrated that putative intact PLs are absent from the inner cortical and nuclear regions of the lens suggesting differential membrane function within the lens.

 
Differential expression of putative lipids in a 16 μm section coated with diaminonaphthalene of the lens of a 2-month old zebrafish. The ions used were 718.69 m/z (purple), 764.51 m/z (orange), 792.75 m/z (blue). The image in the lower right corner is an overlay of 718.69 m/z and 764.51 m/z.
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Differential expression of putative lipids in a 16 μm section coated with diaminonaphthalene of the lens of a 2-month old zebrafish. The ions used were 718.69 m/z (purple), 764.51 m/z (orange), 792.75 m/z (blue). The image in the lower right corner is an overlay of 718.69 m/z and 764.51 m/z.
 
Differential expression of putative lipids in a 16 μm section coated with diaminonaphthalene of the lens of a 2-month old zebrafish. The ions used were 718.69 m/z (purple), 764.51 m/z (orange), 792.75 m/z (blue). The image in the lower right corner is an overlay of 718.69 m/z and 764.51 m/z.
Differential expression of putative lipids in a 16 μm section coated with diaminonaphthalene of the lens of a 2-month old zebrafish. The ions used were 718.69 m/z (purple), 764.51 m/z (orange), 792.75 m/z (blue). The image in the lower right corner is an overlay of 718.69 m/z and 764.51 m/z.
View OriginalDownload Slide
 
Keywords: 551 imaging/image analysis: non-clinical • 583 lipids • 413 aging  
© 2014, The Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Permission to republish any abstract or part of an abstract in any form must be obtained in writing from the ARVO Office prior to publication.
Copyright © 2025 Association for Research in Vision and Ophthalmology.
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