Neuroanatomical tracing combined with in situ hybridization: analysis of gene expression patterns within brain circuits of interest.
Most of our current understanding of brain circuits is based on hodological studies carried out using neuroanatomical tract-tracing. Our aim is to advance one step further by visualizing the functional correlate in a given circuit. In this regard, we believe it is feasible to combine retrograde tracing with fluorescence, non-radioactive in situ hybridization (ISH) protocols. The subsequent detection at the single-cell level of the expression of a given mRNA within retrograde-labeled neurons provides information regarding cellular function. This may be of particular interest when trying to elucidate the performance of brain circuits of interest in animal models of brain diseases. Several combinations of retrograde tracing with either single- and double-ISH are presented here, together with some criteria that influence the selection of the tracer to be used in conjunction with the strong demands of the ISH.
Conte-Perales
Lorena
L
Area de Neurociencias, Centro de Investigación Médica Aplicada (CIMA), Universidad de Navarra and Centro de Investigación en Red sobre Enfermedades Neurodegenerativas (CIBERNED), Pamplona, Spain.
Barroso-Chinea
Pedro
P
Rico
Alberto J
AJ
Gómez-Bautista
Virginia
V
López
Iciar P
IP
Roda
Elvira
E
Wouterlood
Floris G
FG
Lanciego
José L
JL
D000818:Animals; D001923:Brain Chemistry / Q000235:genetics*; D004298:Dopamine / Q000502:physiology; D005456:Fluorescent Dyes; D015870:Gene Expression / Q000502:physiology*; D005917:Globus Pallidus / Q000166:cytology; D005968:Glutamate Decarboxylase / Q000378:metabolism; D000882:Haplorhini; D017403:In Situ Hybridization / Q000379:methods*; D018613:Microscopy, Confocal; D009434:Neural Pathways / Q000502:physiology*; D056347:Neuroanatomical Tract-Tracing Techniques; D009445:Neuroanatomy / Q000379:methods*; D016376:Oligonucleotides, Antisense / Q000138:chemical synthesis; D010477:Perfusion; D012313:RNA / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D013266:Stilbamidines; D020531:Subthalamic Nucleus / Q000166:cytology / Q000502:physiology; D013534:Survival; D016707:Tissue Fixation; D050599:Vesicular Glutamate Transport Protein 2 / Q000096:biosynthesis / Q000235:genetics
C049774:2-hydroxy-4,4'-diamidinostilbene, methanesulfonate salt; D005456:Fluorescent Dyes; D016376:Oligonucleotides, Antisense; C493475:Slc17a6 protein, rat; D013266:Stilbamidines; D050599:Vesicular Glutamate Transport Protein 2; D012313:RNA; D005968:Glutamate Decarboxylase; C401141:glutamate decarboxylase 2; D004298:Dopamine