Event:

Fast synaptic inhibition is one of the main driving forces determining neural output during behavior, yet its exact roles in controlling the activity of individual neurons is challenging to experimentally determine in vivo. The zebra finch is a powerful model system, linking precise premotor neuron output to a learned vocal motor skill – a stereotyped courtship song. Using intracellular and patch recordings, we explored the impact of inhibition on the premotor song network HVC. Local network inhibition gates premotor neuron output in adult song production and has the capacity to control plasticity in a learning dependent manner in juvenile birds. While single-cell electrophysiology is a valuable tool for correlating sub- and suprathreshold neural activity to behavior, it is still difficult to manipulate key genes such as ion channels causally relate intracellular mechanisms to neural function in vivo. To manipulate inhibition at the single-cell level within an otherwise intact network, we developed a method for targeted genetic modification of individual neurons in vivo. We used RNA single-cell electroporation for reliable gene delivery and CRISPR mediated knockout of the GABA-A receptor in neurons of mouse cortex. Individual neurons lacking GABA-A mediated inhibition experienced two distinct phases of adaptation. These studies demonstrate the value of single neuron recordings and manipulations in vivo to reveal unique insights into neural function in awake animals.