Expression of a Soluble Isoform of Cell Adhesion Molecule 1 in the Brain and Its Involvement in Directional Neurite Outgrowth
| Autor: | Man Hagiyama, Keiko B. Kimura, Yoshinori Murakami, Naoki Ichiyanagi, Akihiko Ito |
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| Rok vydání: | 2009 |
| Předmět: |
Gene isoform
Superior cervical ganglion Stromal cell Neurite Blotting Western Intercellular Adhesion Molecule-1 Fluorescent Antibody Technique Biology Pathology and Forensic Medicine Mice In Situ Nick-End Labeling Neurites Animals Immunoprecipitation Protein Isoforms Reverse Transcriptase Polymerase Chain Reaction Cell adhesion molecule Brain Adhesion Immunohistochemistry Molecular biology Coculture Techniques Mice Inbred C57BL Immunoglobulin superfamily Regular Articles |
| Zdroj: | The American Journal of Pathology. 174:2278-2289 |
| ISSN: | 0002-9440 |
| Popis: | Cell adhesion molecule 1 (CADM1), an immunoglobulin superfamily member, is expressed on superior cervical ganglion neurites and mediates cell-cell adhesion by trans-homophilic binding. In addition to the membrane-bound form, we have previously shown that a soluble form (sCADM1) generated by alternative splicing possesses a stop codon immediately downstream of the immunoglobulin-like domain. Here, we demonstrate the presence of sCADM1 in vivo and its possible role in neurite extension. sCADM1 appears to be a stromal protein because extracellular-restricted, but not intracellular-restricted, anti-CADM1 antibody stained stromal protein-rich extract from mouse brains. Murine plasmacytoma cells, P3U1, were modified to secrete sCADM1 fused with either immunoglobulin (Ig)G Fc portion (sCADM1-Fc) or its deletion form that lacks the immunoglobulin-like domain (DeltasCADM1-Fc). When P3U1 derivatives expressing sCADM1-Fc or DeltasCADM1-Fc were implanted into collagen gels, Fc-fused proteins were present more abundantly around the cells. Superior cervical ganglion neurons, parental P3U1, and either derivative were implanted into collagen gels separately, and co-cultured for 4 days. Bodian staining of the gel sections revealed that most superior cervical ganglion neurites turned toward the source of sCADM1-Fc, but not DeltasCADM1-Fc. Furthermore, immunofluorescence signals for sCADM1-Fc and membrane-bound CADM1 were co-localized on the neurite surface. These results show that sCADM1 appears to be involved in directional neurite extension by serving as an anchor to which membrane-bound CADM1 on the neurites can bind. |
| Databáze: | OpenAIRE |
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