Localization of the lens intermediate filament switch by imaging mass spectrometry.
CP49
Filensin
Image segmentation
Imaging mass spectrometry
Lens fiber cell
Lens protein
Vimentin
Journal
Experimental eye research
ISSN: 1096-0007
Titre abrégé: Exp Eye Res
Pays: England
ID NLM: 0370707
Informations de publication
Date de publication:
09 2020
09 2020
Historique:
received:
21
04
2020
revised:
07
06
2020
accepted:
29
06
2020
pubmed:
20
7
2020
medline:
29
1
2021
entrez:
20
7
2020
Statut:
ppublish
Résumé
Imaging mass spectrometry (IMS) enables targeted and untargeted visualization of the spatial localization of molecules in tissues with great specificity. The lens is a unique tissue that contains fiber cells corresponding to various stages of differentiation that are packed in a highly spatial order. The application of IMS to lens tissue localizes molecular features that are spatially related to the fiber cell organization. Such spatially resolved molecular information assists our understanding of lens structure and physiology; however, protein IMS studies are typically limited to abundant, soluble, low molecular weight proteins. In this study, a method was developed for imaging low solubility cytoskeletal proteins in the lens; a tissue that is filled with high concentrations of soluble crystallins. Optimized tissue washes combined with on-tissue enzymatic digestion allowed successful imaging of peptides corresponding to known lens cytoskeletal proteins. The resulting peptide signals facilitated segmentation of the bovine lens into molecularly distinct regions. A sharp intermediate filament transition from vimentin to lens-specific beaded filament proteins was detected in the lens cortex. MALDI IMS also revealed the region where posttranslational myristoylation of filensin occurs and the results indicate that truncation and myristoylation of filensin starts soon after filensin expression increased in the inner cortex. From intermediate filament switch to filensin truncation and myristoylation, multiple remarkable changes occur in the narrow region of lens cortex. MALDI images delineated the boundaries of distinct lens regions that will guide further proteomic and interactomic studies.
Identifiants
pubmed: 32682822
pii: S0014-4835(20)30392-4
doi: 10.1016/j.exer.2020.108134
pmc: PMC7508834
mid: NIHMS1616031
pii:
doi:
Substances chimiques
Cytoskeletal Proteins
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
108134Subventions
Organisme : NEI NIH HHS
ID : P30 EY008126
Pays : United States
Organisme : NEI NIH HHS
ID : R01 EY013462
Pays : United States
Organisme : NEI NIH HHS
ID : U54 EY032442
Pays : United States
Informations de copyright
Copyright © 2020 Elsevier Ltd. All rights reserved.
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