title: Digoxigenylated wheat germ agglutinin visualized with alkaline phosphatase-labeled anti-digoxigenin antibodies--a new, sensitive technique with the potential for single and double tracing of neuronal connections.
abstract: For double tracing experiments, wheat germ agglutinin (WGA) molecules labeled with two different haptens are desirable. In the present report the suitability of digoxigenylated WGA (DIG-WGA) for retrograde tracing was investigated. For this purpose the new tracer was pressure injected into rat brains and the transported DIG-WGA visualized via its digoxigenyl group with an alkaline phosphatase linked anti DIG antibody in permanently stained sections of high quality. With fixatives containing 2.5% glutaraldehyde only few positive cells were found. However, at milder fixation conditions (4% paraformaldehyde, 0.05% glutaraldehyde 0.2% picric acid, 30 min) retrogradely labeled cells were detected with a sensitivity comparable to tetramethylbenzidine protocols for conventional WGA-HRP (horseradish peroxidase) tracing. Preliminary experiments suggest excellent suitability for double labeling.
authors.affiliation: Abteilung für Neuroanatomie, Ruhr-Universität Bochum, F.R.G.
authors.forename: R W
authors.initials: RW
authors.lastname: Veh
mesh_terms: D000469:Alkaline Phosphatase / Q000276:immunology; D000818:Animals; D001921:Brain / Q000166:cytology; D004076:Digoxigenin / Q000276:immunology; D006735:Horseradish Peroxidase; D007150:Immunohistochemistry; D008297:Male; D009434:Neural Pathways / Q000737:chemistry* / Q000276:immunology; D009474:Neurons / Q000737:chemistry* / Q000276:immunology; D009830:Olfactory Bulb / Q000166:cytology; D051381:Rats; D011919:Rats, Inbred Strains; D013194:Staining and Labeling; D018998:Wheat Germ Agglutinin-Horseradish Peroxidase Conjugate; D014909:Wheat Germ Agglutinins
chemical_list: D018998:Wheat Germ Agglutinin-Horseradish Peroxidase Conjugate; D014909:Wheat Germ Agglutinins; D006735:Horseradish Peroxidase; D000469:Alkaline Phosphatase; D004076:Digoxigenin