Glycoconjugate expression in the extracellular matrix of the mouse lung (87.7)

Abstract

Introduction: Glycoconjugates have been implicated in various lung extracellular structures such as the galactose‐binding elastin receptor complex; however, little is known about expression of carbohydrate moieties in lung extracellular matrix (ECM). Methods: To investigate structural implications of glycoconjugate expression in the ECM, we used a panel of 33 biotinylated lectins to label decellularized mouse lungs harvested from 8‐11 week C57/B6 mice. Lungs were decellularized by serial washes through the trachea and pulmonary artery. Standard OCT thin section analysis as well as precision‐cut lung slice (200‐300μm) histology were performed. Lung samples were labeled with lectins (20μg/mL), secondarily labeled with a streptavidin‐fluorescein conjugate and analyzed by fluorescence confocal microscopy. Results: Decellularized mouse lungs showed a lectin‐binding pattern consistent with differential staining of the axial and peripheral connective tissue elements. Some lectins (N=10) demonstrated weak signal interpreted as negative. Others (N=17) stained pleural and subpleural zones suggesting selective staining of the peripheral connective tissue system. Another group (N=7) prominently stained the acinar septae suggesting selective staining of the axial connective tissue system. All lectin labeling was demonstrated at wavelengths that preclude elastin autofluorescence. Conclusions: The differential labeling pattern of the ECM with lectins suggests their utility as monospecific probes of both peripheral and axial connective tissue systems in the decellularized mouse lung.Grant Funding Source: Supported by NIH grant 04925