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Low-affinity nerve growth factor receptor is associated with motoneuron axonal pathways
1Department of Pharmacology, Gunma University, School of Medicine, Maebashi, Japan. hitanka@gpo.kumamoto-u.ac.jp
Abstract:
The unidentified cell-surface antigen recognized by monoclonal antibody M7412 is distributed along motoneuron axonal outgrowth pathways in chicken embryos. To better characterize its role in motoneuron development, the M7412 antigen was purified from chicken embryos by immunoaffinity chromatography. Its N-terminal amino acid sequence corresponded to that predicted for chicken low-affinity nerve growth factor receptor (LNGFR). Experiments were performed to confirm that LNGFR was indeed the antigen recognized by M7412. First, M7412 bound to recombinant chicken LNGFR expressed in mammalian cells. Second, a rabbit serum raised to the purified antigen showed the same staining pattern in chicken embryos as did M7412. Lastly, a novel method for direct detection of nerve growth factor (NGF) bound to its receptors was used to show that in mixed spinal cord cultures, only neurons that expressed M7412 antigen had low-affinity binding sites for NGF. Furthermore, at the subcellular level, M7412 labeling was co-localized with bound NGF. The M7412 antigen is thus chicken LNGFR, whose role in motoneuron outgrowth pathways is discussed.