April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
MALDI Imaging Of Beta And Gamma Crystallins In Bovine And Human Lenses
Author Affiliations & Notes
  • David M. Anderson
    Biochemistry, Vanderbilt University, Nashville, Tennessee
  • Kristie M. Rose
    Biochemistry, Vanderbilt University, Nashville, Tennessee
  • Kevin L. Schey
    Biochemistry, Vanderbilt University, Nashville, Tennessee
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 4749. doi:
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      David M. Anderson, Kristie M. Rose, Kevin L. Schey; MALDI Imaging Of Beta And Gamma Crystallins In Bovine And Human Lenses. Invest. Ophthalmol. Vis. Sci. 2011;52(14):4749.

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

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Abstract

Purpose: : MALDI imaging provides an alternative multiplex imaging modality that depends on the molecular weight of the species present in tissue sections. Thus, intact and modified proteins can be imaged in a single experiment. The purpose of this study is to develop sample preparation methods that allow distributions of beta- and gamma-crystallins and their modified forms to be imaged from bovine and human lens tissue sections.

Methods: : Frozen bovine and human lens sections were obtained at 20 um thickness. Lens tissue was washed with 50% acetonitrile (ACN) and sinapinic acid matrix (50:49.9:0.1 ACN:H2O:TFA) was applied using the Labcyte Portrait 630 spotter. Images were acquired using a Bruker Autoflex III in linear mode with a spatial resolution of 300 µm. Proteins were identified by top down tandem mass spectrometry (both collision induced dissociation and electron transfer dissociation) on-line with HPLC separation of lens homogenates or directly from purified lens crystallins.

Results: : A modified sample preparation protocol allowed images to be produced of beta-and gamma- crystallins from bovine and human lens sections. Gamma-S crystallin showed an outer cortical localization while other gamma crystallins were observed in the nucleus. Partially truncated beta crystallins were distributed in an outer nuclear ring.

Conclusions: : MALDI imaging methods have been expanded to include not only alpha-crystallin, but now beta- and gamma-crystallins in lens tissues. Images can be used to detect age-related modifications and their spatial localization.

Keywords: crystallins • imaging/image analysis: non-clinical • proteomics 
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