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Journal of Histochemistry and Cytochemistry, Vol. 51, 1331-1341, October 2003, Copyright © 2003, The Histochemical Society, Inc.

ARTICLE

Perlecan, the Multidomain Heparan Sulfate Proteoglycan of Basement Membranes, Is also a Prominent Component of the Cartilaginous Primordia in the Developing Human Fetal Spine

James Melrosea, Susan Smitha, Peter Ghosha, and John Whitelockb
a The Institute of Bone and Joint Research, The University of Sydney, at the Royal North Shore Hospital of Sydney, St. Leonards, Australia
b Biomics Research Services, Homebush, Australia

Correspondence to: James Melrose, Raymond Purves Bone and Joint Research Laboratories, Inst. of Bone and Joint Research, Level 5, The University Clinic, Building B26, The Royal North Shore Hospital, St. Leonards NSW 2065, Australia. E-mail: jmelrose@mail.usyd.edu.au

The aim of this study was to localize perlecan in human fetal spine tissues. Human fetal spines (12–20 weeks; n=6) were fixed in either Histochoice or 10% neutral buffered formalin, routinely processed, paraffin-embedded, and 4-µm sagittal sections were cut and stained with toluidine blue, H&E, and von Kossa. Perlecan, types I, II, IV, and X collagen, CD-31, aggrecan core protein, and native and {Delta}-HS 4, 5 hexuronate stub epitopes were immunolocalized. Toluidine blue staining visualized the cartilaginous vertebral body (VB) rudiments and annular lamellae encompassing the nucleus pulposus (NP). Von Kossa staining identified the VB primary center of ossification. Immunolocalization of type IV collagen, CD-31, and perlecan delineated small blood vessels in the outer annulus fibrosus (AF) and large canals deep within the VBs. Perlecan and type X collagen were also prominently expressed by the hypertrophic vertebral growth plate chondrocytes. Aggrecan was extracellularly distributed in the intervertebral disk (IVD) with intense staining in the posterior AF. Notochordal tissue stained strongly for aggrecan but negatively for perlecan and types I and II collagen. Type I collagen was prominent in the outer AF and less abundant in the NP, while type II collagen was localized throughout the IVD and VB. The immunolocalization patterns observed indicated key roles for perlecan in vasculogenic, chondrogenic, and endochondral ossification processes associated with spinal development.

(J Histochem Cytochem 51:1331–1341, 2003)

Key Words: perlecan, HS proteoglycan, human fetal cartilage, development, vertebral growth plate, hypertrophic growth plate, chondrocytes


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