Development of an antibody against a 40,000 mol. wt brain injury-derived neurotrophic peptide-binding protein and identification of a 40,000 mol. wt brain injury-derived neurotrophic peptide-binding protein in hippocampal neurons.
Brain injury-derived neurotrophic peptide is a 13-amino acid peptide derived from a 15,000 mol. wt neurotrophic factor released from sites of mechanical injury in neonatal rat brain. This peptide promotes survival of septal cholinergic neurons and mesencephalic dopaminergic neurons, and protects hippocampal neurons from glutamate-induced neurotoxicity. In this study, we have developed a monoclonal antibody against a brain injury-derived neurotrophic peptide-binding protein by immunizing mice with septal synaptosomes from five-week-old rat brain. Monoclonal antibodies were screened for inhibition of the binding of a 125I-labeled analogue of brain injury-derived neurotrophic peptide to rat brain synaptosomes. The monoclonal antibody 6A22 suppressed the biological activity of brain injury-derived neurotrophic peptide and abolished the protective effect of the neurotrophic peptide against glutamate-induced neurotoxicity. This monoclonal antibody recognized a 40,000 mol. wt brain injury-derived neurotrophic peptide-binding protein, which was also identified by cross-linking experiments. Immunohistochemical studies showed that the 6A22 antibody bound to the cell surfaces of a subpopulation (about 60%) of hippocampal neurons in culture. These results are consistent with the possibility that the 40,000 mol. wt protein belongs to brain injury-derived neurotrophic peptide receptors.
Hama
T
T
Mitsubishi Kasei Institute of Life Sciences, 11 Minamiooya, Machida-shi, 194-8511, Tokyo, Japan. hama@libra.ls.m-kagaku.co.jp
Maruyama
M
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D016923:Cell Death / Q000187:drug effects / Q000502:physiology; D003432:Cross-Linking Reagents / Q000378:metabolism; D003470:Culture Media / Q000494:pharmacology; D005333:Fetus / Q000166:cytology; D018698:Glutamic Acid / Q000633:toxicity; D006624:Hippocampus / Q000166:cytology; D006825:Hybridomas; D007457:Iodine Radioisotopes; D051379:Mice; D008970:Molecular Weight; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000138:chemical synthesis / Q000276:immunology; D009447:Neuroblastoma; D009474:Neurons / Q000737:chemistry* / Q000166:cytology; D011233:Precipitin Tests; D011485:Protein Binding / Q000502:physiology; D051381:Rats; D013574:Synaptosomes / Q000737:chemistry; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D003432:Cross-Linking Reagents; D003470:Culture Media; D007457:Iodine Radioisotopes; D009419:Nerve Tissue Proteins; C102215:brain injury-derived neurotrophic peptide; D018698:Glutamic Acid