Novel Single-Neuron-Tracing Method Using Sindbis Viral Vectors
Abstract: Novel methods for Golgi-stain-like labeling of neurons have been developed by applying molecular biological technique. Using replication-deficient Sindbis viral vectors expressing membrane-targeted green fluorescent protein (GFP) or monomeric red fluorescent protein (mRFP), we visualized the neuronal processes of the infected neurons completely. Furthermore, when sufficiently diluted Sindbis virus solution was injected into the brain, the entire axonal arborization of the infected neurons was recovered at a single cell level, giving new insights into neuronal networks. Since neuronal processes, axons and dendrites, are key structures of neuronal information processing, the Sindbis viral vectors introduced here are useful for the morphological analysis of neuronal networks and thus helpful for understanding the functional design of the nervous system.
Key words: single neuron analysis, Sindbis virus, Golgi-like labeling, thalamo-cortical projection, motor thalamic nuclei