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1.
Exp Eye Res ; 201: 108326, 2020 12.
Artículo en Inglés | MEDLINE | ID: mdl-33147472

RESUMEN

The Descemet's membrane (DM) and the lens capsule (LC) are two ocular basement membranes (BMs) that are essential in maintaining stability and structure of the cornea and lens. In this study, we investigated the proteomes and biomechanical properties of these two materials to uncover common and unique properties. We also screened for possible protein changes during diabetes. LC-MS/MS was used to determine the proteomes of both BMs. Biomechanical measurements were conducted by atomic force microscopy (AFM) in force spectroscopy mode, and complemented with immunofluorescence microscopy. Proteome analysis showed that all six existing collagen IV chains represent 70% of all LC-protein, and are thus the dominant components of the LC. The DM on the other hand is predominantly composed of a single protein, TGF-induced protein, which accounted for around 50% of all DM-protein. Four collagen IV-family members in DM accounted for only 10% of the DM protein. Unlike the retinal vascular BMs, the LC and DM do not undergo significant changes in their protein compositions during diabetes. Nanomechanical measurements showed that the endothelial/epithelial sides of both BMs are stiffer than their respective stromal/anterior-chamber sides, and both endothelial and stromal sides of the DM were stiffer than the epithelial and anterior-chamber sides of the LC. Long-term diabetes did not change the stiffness of the DM and LC. In summary, our analyses show that the protein composition and biomechanical properties of the DM and LC are different, i.e., the LC is softer than DM despite a significantly higher concentration of collagen IV family members. This finding is unexpected, as collagen IV members are presumed to be responsible for BM stiffness. Diabetes had no significant effect on the protein composition and the biomechanical properties of both the DM and LC.


Asunto(s)
Membrana Basal/metabolismo , Córnea/metabolismo , Lámina Limitante Posterior/metabolismo , Proteínas del Ojo/metabolismo , Cápsula del Cristalino/metabolismo , Anciano , Membrana Basal/citología , Cromatografía Liquida , Lámina Limitante Posterior/citología , Elasticidad , Femenino , Humanos , Cápsula del Cristalino/citología , Masculino , Microscopía de Fuerza Atómica , Persona de Mediana Edad , Espectrometría de Masas en Tándem
2.
FEBS J ; 291(3): 477-488, 2024 02.
Artículo en Inglés | MEDLINE | ID: mdl-37984833

RESUMEN

Basement membranes are among the most widespread, non-cellular functional materials in metazoan organisms. Despite this ubiquity, the links between their compositional and biophysical properties are often difficult to establish due to their thin and delicate nature. In this article, we examine these features on a molecular level by combining results from proteomics, elastic, and nanomechanical analyses across a selection of human basement membranes. Comparing results between these different membranes connects certain compositional attributes to distinct nanomechanical signatures and further demonstrates to what extent water defines these properties. In all, these data underline BMs as stiff yet highly elastic connective tissue layers and highlight how the interplay between composition, mechanics and hydration yields such exceptionally adaptable materials.


Asunto(s)
Laminina , Humanos , Animales , Membrana Basal/química , Microscopía de Fuerza Atómica , Laminina/análisis
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