Multiple neuroanatomical tracing in primates.
The present report deals with a multiple tract-tracing procedure in non-human primates enabling the simultaneous visualization of retrogradely transported Fluoro-Gold (FG) and cholera toxin B subunit (CTB) in combination with anterogradely transported biotinylated dextran amine (BDA). Two issues have played key roles on the achievement of this reliable procedure: first, the recent development of a commercial antiserum against FG that allows us to convert the original fluorescent signal of this dye in a permanent precipitate via standard peroxidase-anti-peroxidase methods; second, the introduction of the novel peroxidase substrate Vector(R) VIP (V-VIP), resulting in a purple precipitate. The combination of these neuroanatomical tracers in one and the same histological section opens a possibility for the permanent visualization of the convergence of inputs from a particular brain area onto identified, two different subsets of projection cells of another area. Furthermore, this combination of three tracers emerges as a powerful technical tool for obtaining broad amounts of complementary data regarding the monkey brain connectivity, thus significantly reducing the number of animals needed to complete a particular study.
Lanciego
J L
JL
Departamento de Anatomía, Facultad de Medicina, Universidad de Navarra, Pamplona, Spain. jlanciego@unav.es
Luquin
M R
MR
Guillén
J
J
Giménez-Amaya
J M
JM
D000818:Animals; D001692:Biological Transport; D001710:Biotin / Q000031:analogs & derivatives* / Q000032:analysis; D001931:Brain Mapping / Q000379:methods*; D002552:Cerebral Ventricles / Q000502:physiology; D002772:Cholera Toxin / Q000032:analysis*; D003911:Dextrans / Q000032:analysis*; D005456:Fluorescent Dyes / Q000032:analysis*; D007124:Immunoenzyme Techniques; D008252:Macaca fascicularis; D008297:Male; D009434:Neural Pathways / Q000502:physiology; D013238:Stereotaxic Techniques; D013266:Stilbamidines
C049774:2-hydroxy-4,4'-diamidinostilbene, methanesulfonate salt; D003911:Dextrans; D005456:Fluorescent Dyes; D013266:Stilbamidines; C076397:biotinylated dextran amine; D001710:Biotin; D002772:Cholera Toxin