Lentivirus-based genetic manipulations of cortical neurons and their optical and electrophysiological monitoring in vivo

摘要: It is becoming increasingly clear that single cortical neurons encode complex and behaviorally relevant signals, but efficient means to study gene functions in small networks vivo are still lacking. Here, we establish a method for genetic manipulation subsequent phenotypic analysis of individual vivo. First, lentiviral vectors used neuron-specific delivery from alpha-calcium/calmodulin-dependent protein kinase II or Synapsin I promoters, optionally combination with knockdown by U6 promoter-driven expression short-interfering RNAs. Second, the at level cells carried out using two-photon microscopy-based techniques: high-resolution time-lapse imaging monitor structural dynamics dendritic spines axonal projections, whereas cellular response properties analyzed electrophysiologically microscopy directed whole-cell recordings. This approach ideally suited intact brain.