A sequential protocol combining dual neuroanatomical tract-tracing with the visualization of local circuit neurons within the striatum.
We describe here an experimental approach designed to aid in the identification of complex brain circuits within the rat corpus striatum. Our aim was to characterize in a single section (i) striatal thalamic afferents, (ii) striatopallidal projection neurons and (iii) striatal local circuit interneurons. To this end, we have combined anterograde tracing using biotinylated dextran amine and retrograde neuroanatomical tracing with Fluoro-Gold. This dual tracing protocol was further implemented with the visualization of different subpopulations of striatal interneurons. The subsequent use of three different peroxidase substrates enabled us to unequivocally detect structures that were labeled within a three-color paradigm.
Gonzalo
N
N
Departamento de Anatomía, Facultad de Medicina, Universidad de Navarra, Pamplona, Spain.
Moreno
A
A
Erdozain
M A
MA
García
P
P
Vázquez
A
A
Castle
M
M
Lanciego
J L
JL
D015100:3,3'-Diaminobenzidine; D000818:Animals; D001360:Avidin / Q000031:analogs & derivatives*; D001369:Axons / Q000378:metabolism / Q000648:ultrastructure; D001710:Biotin / Q000031:analogs & derivatives*; D064032:Calbindin 2; D048429:Cell Size / Q000502:physiology; D002795:Choline O-Acetyltransferase / Q000378:metabolism; D003712:Dendrites / Q000378:metabolism / Q000648:ultrastructure; D003911:Dextrans; D005260:Female; D005456:Fluorescent Dyes; D006735:Horseradish Peroxidase; D007150:Immunohistochemistry / Q000379:methods*; D007395:Interneurons / Q000166:cytology* / Q000378:metabolism; D015335:Molecular Probes; D009252:NADPH Dehydrogenase / Q000378:metabolism; D017072:Neostriatum / Q000166:cytology* / Q000378:metabolism; D009434:Neural Pathways / Q000166:cytology* / Q000378:metabolism; D009532:Nickel; D009580:Nitroblue Tetrazolium; D010320:Parvalbumins / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D064030:S100 Calcium Binding Protein G / Q000378:metabolism; D013266:Stilbamidines
C049774:2-hydroxy-4,4'-diamidinostilbene, methanesulfonate salt; D064032:Calbindin 2; D003911:Dextrans; D005456:Fluorescent Dyes; D015335:Molecular Probes; D010320:Parvalbumins; D064030:S100 Calcium Binding Protein G; D013266:Stilbamidines; C083986:avidin-horseradish peroxidase complex; C076397:biotinylated dextran amine; D001360:Avidin; D009580:Nitroblue Tetrazolium; D015100:3,3'-Diaminobenzidine; D001710:Biotin; D009532:Nickel; D006735:Horseradish Peroxidase; D009252:NADPH Dehydrogenase; D002795:Choline O-Acetyltransferase