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Localization and quantification of the insulin-like growth factor-1 receptor in mouse articular cartilage by confocal laser scanning microscopy

PJ Verschure, J van Marle, LA Joosten and WB Van den Berg

Department of Rheumatology, University Hospital Nijmegen, The Netherlands.

Insulin-like growth factor-1 (IGF-1) is an essential anabolic growth factor in the regulation of cartilage metabolism and exerts its effects by binding to the IGF-1 Type 1 receptor on the chondrocyte membrane. We have localized and quantified in situ IGF-1 receptor expression in intact articular cartilage of normal mice. The IGF-1 receptor was detected immunohistochemically with antibodies to the IGF-1 receptor and visualized with conventional light microscopy and confocal laser scanning microscopy (CLSM). CLSM analysis enabled us to distinguish IGF- 1 receptor immunoreactivity on the chondrocyte cell membrane from intracellular staining. We have established two approaches to quantify in confocal images low levels of fluorescence intensity of the immunolocalized IGF-1 receptor at the chondrocyte membrane, i.e., mean pixel measurement and area measurement. The majority of IGF-1 receptor fluorescence intensity was localized on chondrocytes in the middle and deeper zones of cartilage, whereas chondrocytes in the surface zone exhibited negligible fluorescence. The variable distribution of IGF-1 receptor in chondrocytes of articular cartilage suggests that effects of IGF-1 on chondrocytes may be distinctly heterogeneous in the different mouse articular cartilage zones.

Volume 42, Issue 6, pp. 765-773, 06/01/1994
Copyright © 1994 by The Histochemical Society


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