Visualization of dendritic morphology of cortical projection neurons by retrograde axonal tracing

Currently there is no reliable retrograde tracing technique for visualization of dendritic morphologies of projection neurons. Here we describe a simple and efficient method that can be used to label neurons in Golgi-like fashion. The approach relies on activity-dependent uptake of tracer. For this purpose we inject the glutamate receptor agonist N-methyl-D,L-aspartic acid (NMDA) at the tracer injection site to massively stimulate neurons and to thereby promote uptake of biocytin or biotinylated dextran amine (BDA) by axon terminals. The results show that co-injections of NMDA/biocytin and NMDA/BDA into the extrastriate lateromedial area (LM) of rat visual cortex labels large numbers of neurons in area 17 in Golgi-like fashion. Similarly injections of the lateral geniculate nucleus (LGN) lead to Golgi-like labeling of corticogeniculate neurons in area 17. The distribution of labeled neurons is highly topographic. In addition the method allows excellent preservation of ultrastructure, indicating that this approach is useful for determining the organization of neuronal circuits within the central nervous system.