Systematic screening and identification of antigens recognized by monoclonal antibodies raised against the developing lateral olfactory tract.
During development, olfactory bulb axons navigate a complex microenvironment composed of myriad molecules to construct a bundle called the lateral olfactory tract. The axons themselves also express thousands of different molecules. In the present study, we produced and characterized six monoclonal antibodies that label the lateral olfactory tract and its surroundings in a unique pattern. The labeling profiles suggested that the antigen molecules recognized by each antibody are heterogeneously distributed around the developing lateral olfactory tract. We developed an efficient screening method to identify the antigen molecules by combining expression of a cDNA library in COS-7 cells and the subsequent immunohistochemical staining of the cells. The systematic screening successfully identified specific cDNA clones for all of the monoclonal antibodies, which highly probably coded for the antigen molecules, and therefore unveiled the molecular nature of local components that embrace the developing lateral olfactory tract in mice.
Kawasaki
Takahiko
T
Division of Brain Function, National Institute of Genetics, Graduate University for Advanced Studies, Yata 1111, Mishima 411-8540, Japan.
Takagi
Yoshiko
Y
Yamatani
Hitoshi
H
Hirata
Tatsumi
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity / Q000502:physiology; D000941:Antigens / Q000276:immunology*; D001369:Axons / Q000378:metabolism; D015153:Blotting, Western / Q000379:methods; D019556:COS Cells; D002352:Carrier Proteins / Q000378:metabolism; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D006224:Cricetinae; D004622:Embryo, Mammalian; D005260:Female; D019922:GAP-43 Protein / Q000378:metabolism; D005820:Genetic Testing / Q000379:methods; D007150:Immunohistochemistry / Q000379:methods; D036341:Intercellular Signaling Peptides and Proteins; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D008813:Mice, Inbred ICR; D009419:Nerve Tissue Proteins / Q000378:metabolism; D019006:Neural Cell Adhesion Molecules / Q000378:metabolism; D009474:Neurons / Q000378:metabolism; D009833:Olfactory Pathways / Q000166:cytology / Q000254:growth & development / Q000276:immunology* / Q000378:metabolism; D011247:Pregnancy; D016601:RNA-Binding Proteins / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D011980:Receptors, Progesterone / Q000378:metabolism; D013116:Spinal Cord / Q000378:metabolism; D014162:Transfection / Q000379:methods
D000911:Antibodies, Monoclonal; D000941:Antigens; D002352:Carrier Proteins; D019922:GAP-43 Protein; D036341:Intercellular Signaling Peptides and Proteins; D008565:Membrane Proteins; C451223:NDFIP1 protein, human; C499188:Ndfip1 protein, mouse; D009419:Nerve Tissue Proteins; D019006:Neural Cell Adhesion Molecules; C486424:Pgrmc1 protein, rat; D016601:RNA-Binding Proteins; D011980:Receptors, Progesterone; C489914:Rtn1 protein, mouse