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Bronchial Branching Correlates with Specific Glycosidase Activity, Extracellular Glycosaminoglycan Accumulation, TGFß₂, and IL-1 Localization During Chick Embryo Lung Development

M. Calvitti¹, T. Baroni¹, C. Calastrini, C. Lilli, E. Caramelli, E. Becchetti, P. Carinci, L. Vizzotto and G. Stabellini

Experimental Medicine and Biochemistry Science Department, University of Perugia (MC,TB,CL,EB); Morphology Embriology Department, Section of Histology and Embryology, University of Ferrara (CC); Histology and Embryology Institute and Center of Molecular Genetics, University of Bologna (EC,PC); Human Anatomy Department–Polo of Vialba, University of Milano (LV); and Human Anatomy Department–L.I.T.A of Segrate, State University of Milano (GS), Milan, Italy

Correspondence to: Giordano Stabellini, Dipartimento di Anatomia Umana, Via Mangiagalli 31, 20133 Milano, Italy. E-mail: giordano.stabellini{at}unimi.it

During organ differentiation, cell–extracellular matrix (ECM) interactions are required. The components of the ECM, such as glycosaminoglycans, fibronectin, laminin, and collagens, change in relation to cytokine and enzyme activity. Moreover, glycosaminoglycans (GAGs) are components of the ECM that play an important role in both cytokine regulation and cell activities. In this work we studied the accumulation of hyaluronic acid and chondroitin sulfate and heparan sulfate proteoglycans (PGs), ß-N-acetyl-D-glucosaminidase activity, the presence of transforming growth factor ß₂ (TGFß₂), and interleukin-1 (IL-1), and the localization of fibronectin, laminin, and collagen I and IV during the early stages of chick embryo lung development. We also determined the levels of hyaluronic acid, chondroitin sulfate, dermatan sulfate, and heparan sulfate GAGs and the activity of ß-N-acetyl-D-glucosaminidase with biochemical methods. Our data show that ß-N-acetyl-D-glucosaminidase activity increases in each cell, especially in the epithelial growth front at the emergence of each bronchial bud, where hyaluronic acid and IL-1 are located in the surrounding mesenchymal areas. Chondroitin sulfate and heparan sulfate PGs, fibronectin, laminin, and collagen I and IV are evident in the area near the basal membrane along the sides where the forming structures are stabilized. Biochemical data show that ß-N-acetyl-D-glucosaminidase activity increases in cells during lung development and is related to GAG decrease and to modifications of the nonsulfated/sulfated GAG ratio. These modifications could change cytokine activity and play an important role in bronchial branching development. (J Histochem Cytochem 52:325–334, 2004)

Key Words: bronchial branching • chondroitin sulfate • proteoglycans • heparan sulfate proteoglycans • interleukin-1 • transforming growth factor ß₂ • laminin • fibronectin • collagen I • collagen IV

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