title: Integrin alpha 2 beta 1 mediates interactions between developing embryonic retinal cells and collagen.
abstract: In the developing nervous system, the extracellular matrix provides a source of extrinsic cues to guide determination of cell fate, neuroblast migration, axon outgrowth and synapse formation. In the neural retina, undifferentiated neuroepithelial precursor cells contact extracellular matrix that contains multiple collagen types. Collagens have been shown to support retinal cell adhesion and neurite outgrowth, but the integrin receptors mediating neuronal responses are not understood. Here we provide evidence that integrin alpha 2 beta 1 acts as a collagen receptor in the developing avian retina and examine its expression pattern. Using a recently described monoclonal antibody, MEP-17, alpha 2 protein was detected in the developing retina by immunofluorescence in tissue sections and dissociated cells, and by immunoprecipitation. At embryonic day 4 (E4), when the majority of retinal cells are undifferentiated neuroepithelial cells, alpha 2 immunoreactivity in sections was widespread and about half of cells dissociated in culture were alpha 2 positive. At E6, after the retinal ganglion cell layer had differentiated, immunoreactivity in sections decreased in the central, more developed portion of the retina and 25% of dissociated cells were alpha 2 positive. E6 retinal ganglion cells, identified by neurofilament immunoreactivity, did not express detectable alpha 2 immunoreactivity. Immunoprecipitation experiments using E6 extracts demonstrated that the alpha 2 subunit was paired with the beta 1 integrin subunit. By E12, alpha 2 immunoreactivity in sections was confined to the extreme peripheral retina, although the antigen may be masked since expression levels comparable to or slightly higher than E6 could be detected in dissociated cells and extracts. By employing function blocking antibodies, it was shown that alpha 2 beta 1 integrin is necessary for cell adhesion and process outgrowth by embryonic retinal cells on collagens I and IV. Although alpha 2 expression continued through E12, alpha 2 activity was down regulated with increasing embryonic age, since alpha 2-dependent adhesion and outgrowth declined. These data suggest a role for alpha 2 beta 1 in neuroepithelial cell interactions with collagen rather than for axon extension by retinal ganglion cells.
authors.affiliation: Neuroscience Research Institute, University of California, Santa Barbara 93106, USA.
authors.forename: A D
authors.initials: AD
authors.lastname: Bradshaw
authors.forename: K M
authors.initials: KM
authors.lastname: McNagny
authors.forename: D B
authors.initials: DB
authors.lastname: Gervin
authors.forename: G M
authors.initials: GM
authors.lastname: Cann
authors.forename: T
authors.initials: T
authors.lastname: Graf
authors.forename: D O
authors.initials: DO
authors.lastname: Clegg
mesh_terms: D000818:Animals; D002448:Cell Adhesion / Q000502:physiology; D002642:Chick Embryo; D003094:Collagen / Q000502:physiology*; D005109:Extracellular Matrix / Q000502:physiology; D005455:Fluorescent Antibody Technique; D005865:Gestational Age; D019012:Integrin beta1 / Q000032:analysis / Q000502:physiology*; D016023:Integrins / Q000502:physiology*; D011233:Precipitin Tests; D038981:Receptors, Collagen; D012160:Retina / Q000737:chemistry / Q000166:cytology / Q000196:embryology*; D013234:Stem Cells / Q000502:physiology*
chemical_list: D019012:Integrin beta1; D016023:Integrins; D038981:Receptors, Collagen; D003094:Collagen