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Journal of Histochemistry and Cytochemistry, Vol. 50, 1313-1324, October 2002, Copyright © 2002, The Histochemical Society, Inc.

ARTICLE

Latrunculin and Cytochalasin Decrease Chondrocyte Matrix Retention

Ghada A. Nofala and Cheryl B. Knudsona
a Departments of Biochemistry and Pathology, Rush Medical College, Rush-Presbyterian–St Luke's Medical Center, Chicago, Illinois

Correspondence to: Cheryl B. Knudson, Dept. of Biochemistry, Rush Medical College, 1653 W. Congress Parkway, Chicago, IL 60612. E-mail: cknudson@rush.edu

The proteoglycan-rich extracellular matrix (ECM) directly associated with the cells of articular cartilage is anchored to the chondrocyte plasma membrane via interaction with the hyaluronan receptor CD44. The cytoplasmic tail of CD44 interacts with the cortical cytoskeleton. The objective of this study was to determine the role of the actin cytoskeleton in CD44-mediated matrix assembly by chondrocytes and cartilage matrix retention and homeostasis. Adult bovine articular cartilage tissue slices and isolated chondrocytes were treated with latrunculin or cytochalasin. Tissues were processed for histology and chondrocytes were examined for CD44 expression and pericellular matrix assembly. Treatments that disrupt the actin cytoskeleton reduced chondrocyte pericellular matrix assembly and the retention of proteoglycan within cartilage explants. There was enhanced detection of a neoepitope resulting from proteolysis of aggrecan. Cytoskeletal disruption did not reduce CD44 expression, as monitored by flow cytometry, but detergent extraction of CD44 was enhanced and hyaluronan binding was decreased. Thus, disruption of the cytoskeleton reduces the anchorage of CD44 in the chondrocyte membrane and the capacity of CD44 to bind its ligand. The results suggest that cytoskeletal disruption within cartilage uncouples chondrocytes from the matrix, resulting in altered metabolism and deleterious changes in matrix structure. (J Histochem Cytochem 50:1313–1323, 2002)

Key Words: cartilage, chondrocytes, proteoglycans, hyaluronan, CD44, latrunculin, cytochalasin, matrix assembly


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