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Phylogenetic conservation of the cell-type-specific Lan3-2 glycoepitope in Caenorhabditis elegans

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Abstract

The biological function of a cell-type-specific glycosylation of an adhesion molecule belonging to the L1CAM immunoglobulin superfamily was previously determined in the nervous system of the embryonic leech, Hirudo medicinalis. The Lan3-2 glycoepitope is a surface marker of sensory afferent neurons and is required for their appropriate developmental collateral branching and synaptogenesis in the CNS. The chemical structure of the Lan3-2 glycoepitope consists of β-(1,4)-linked mannopyranose. Here, we show the conservation of the cell-type-specific expression of this mannose polymer in Caenorhabditis elegans. The Lan3-2 glycoepitope is expressed on the cell surface of a subset of dissociated embryonic neurons and, in the adult worm, by the pharyngeal motor neuron, M5, and the chemosensory afferents, the amphids. Additionally, the vulval epithelium expresses the Lan3-2 glycoepitope in late L4 larvae and in adult hermaphrodites. To investigate proteins carrying this restrictively expressed glycoepitope, worm extract was immunoaffinity purified with Lan3-2 monoclonal antibody and Western blotted. A polyclonal antibody reactive with the cytoplasmic tail of LAD-1/SAX-7, a C. elegans member of the L1CAM family, recognizes a 270 kDa protein band while Lan3-2 antibody also recognizes a 190 kDa glycoform, its putative Lan3-2 ectodomain. Thus, in C. elegans, as in leech, the Lan3-2 epitope is located on a L1CAM homologue. The cell-type-specific expression of the Lan3-2 glycoepitope shared by leech and C. elegans will be useful for understanding how cell-type-specific glycoepitopes mediate cell–cell interactions during development.

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Acknowledgments

We thank Dr. Martin Chalfie for helping with the identification of Lan3-2 stained neurons. We appreciate generous gifts of sqv1–8 mutants from Dr. Horvitz and Dr. Hwang and LAD-1 antibody from Dr. Lihsia Chen. We thank Drs. Seth Hootman, Joseph Leykam, and Miriam B. Goodman for critically reading the manuscript. This work was supported by National Institute of Health Grant NS5117. Robert O'Hagan was supported by a Predoctoral Fellowship from Howard Hughes Medical Institute.

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Author notes

  1. Harper C. VanSteenhouse

    Present address: Program in Neuroscience, University of California at San Diego, La Jolla, CA, 92093, USA

  2. Zachary A. Horton

    Present address: Michigan State University Grand Rapids Medical Education and Research Center, Grand Rapids, MI, 49503, USA

  3. Robert O’Hagan

    Present address: Department of Genetics, Rutgers University, Piscataway, NJ, 08854, USA

Authors and Affiliations

  1. Department of Physiology, Michigan State University, East Lansing, MI, 48824, USA

    Harper C. VanSteenhouse, Zachary A. Horton, Mei-Hui Tai & Birgit Zipser

  2. Department of Biological Sciences, Columbia University, New York, NY, 10027, USA

    Robert O’Hagan

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  1. Harper C. VanSteenhouse
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Correspondence to Birgit Zipser.

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Communicated by D.A. Weisblat

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VanSteenhouse, H.C., Horton, Z.A., O’Hagan, R. et al. Phylogenetic conservation of the cell-type-specific Lan3-2 glycoepitope in Caenorhabditis elegans . Dev Genes Evol 220, 77–87 (2010). https://doi.org/10.1007/s00427-010-0330-8

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