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Topographical variation in the distributions of versican, aggrecan and perlecan in the foetal human spine reflects their diverse functional roles in spinal development

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Abstract

We evaluated the immunohistochemical distribution of three major proteoglycans of cartilage, i.e., aggrecan, versican and perlecan vis-a-vis collagens I and II in the developing human spine of first-trimester foetuses. Aggrecan and perlecan were prominently immunolocalised in the cartilaginous vertebral body rudiments and to a lesser extent within the foetal intervertebral disc. In contrast, versican was only expressed in the developing intervertebral disc interspace. Using domain-specific monoclonal antibodies against the various modules of versican, we discovered the V0 isoform as the predominant form present. Versican immunolocalisations conducted with antibodies directed to epitopes in its N and C termini and GAG-α and GAG-β core protein domains provided evidence that versican in the nucleus pulposus was either synthesised devoid of a G3 domain or this domain was proteolytically removed in situ. The V0 versican isoform was localised with prominent fibrillar components in the annular lamellae of the outer annulus fibrosus. Perlecan was a notable pericellular proteoglycan in the annulus fibrosus and nucleus pulposus but poorly immunolocalised in the marginal tissues of the developing intervertebral disc, apparently delineating the intervertebral disc–vertebral body interface region destined to become the cartilaginous endplate in the mature intervertebral disc. The distribution of collagens I and II in the foetal spine was mutually exclusive with type I present in the outer annulus fibrosus, marginal tissues around the vertebral body rudiment and throughout the developing intervertebral disc, and type II prominent in the vertebral rudiment, absent in the outer annulus fibrosus and diffusely distributed in the inner annulus fibrosus and nucleus pulposus. Collectively, our findings suggest the existence of an intricate and finely balanced interplay between various proteoglycans and collagens and the spinal cell populations which synthesise and assemble these components during spinal development.

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Acknowledgments

This study was supported by NHMRC Project grant 512167. We would like to thank Prof Firoz Rahemtulla, University of Alabama, Birmingham, USA for the kind gift of the versican 5D5 Mab used in this study.

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Authors and Affiliations

  1. Raymond Purves Bone and Joint Research Laboratories, Institute of Bone and Joint Research, Kolling Institute of Medical Research, The Royal North Shore Hospital, University of Sydney, Level 10, Building B6, St. Leonards, NSW, 2065, Australia

    Susan M. Smith, Christopher B. Little & James Melrose

  2. Graduate School of Biomedical Engineering, University of NSW, Kensington, NSW, 2052, Australia

    John M. Whitelock

  3. Department of Pathology, Anatomy and Cell Biology, The Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA, USA

    Renato V. Iozzo

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  1. Susan M. Smith
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  5. James Melrose
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Correspondence to James Melrose.

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Smith, S.M., Whitelock, J.M., Iozzo, R.V. et al. Topographical variation in the distributions of versican, aggrecan and perlecan in the foetal human spine reflects their diverse functional roles in spinal development. Histochem Cell Biol 132, 491–503 (2009). https://doi.org/10.1007/s00418-009-0623-z

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