Selective immunocytochemical staining of mitral cells in rat olfactory bulb with affinity purified antibodies against N-acetyl-aspartyl-glutamate.
Polyclonal antibodies were raised in rabbits against N-acetyl-aspartyl-glutamate (NAAG) coupled to bovine serum albumin (BSA) with carbodiimide and were purified by affinity chromatography sequentially over BSA-agarose and NAAG-agarose resins. Solid-phase RIA revealed a distinct pattern of specificity of the antibodies for N-acetylated acidic peptides, with highest signal obtained for NAAG, and essentially no immunoreactivity demonstrable for aspartate or glutamate. Coronal sections through carbodiimide-fixed rat olfactory bulb were incubated with the purified antiserum and antigen localization visualized by the avidin-biotin peroxidase techniques. Immunoreactivity was restricted to the mitral cells, the major excitatory projection neurons of the lateral olfactory tract, a putative glutamatergic pathway. Immunoreactivity was selectively blocked by preincubation of the antibody with 1 microgram/ml of NAAG-BSA. These results demonstrate a specific neuronal localization of NAAG-like immunoreactivity and support the candidacy of NAAG as a neurotransmitter of the lateral olfactory tract.
D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000302:isolation & purification*; D004151:Dipeptides / Q000276:immunology*; D006651:Histocytochemistry; D007120:Immunochemistry; D008297:Male; D009830:Olfactory Bulb / Q000166:cytology / Q000276:immunology*; D051381:Rats; D011919:Rats, Inbred Strains; D013194:Staining and Labeling
D000906:Antibodies; D004151:Dipeptides; C027172:isospaglumic acid