Transfection via whole-cell recording in vivo: Bridging single-cell physiology, genetics and connectomics

Ede A Rancz (1) , Kevin M. Franks (1) , Martin K. Schwarz (1) , Bruno Pichler (1) , Andreas T. Schaefer (1) , Troy W Margrie (1)

1 Neurophysiology

Single cell genetic manipulation is expected to significantly advance the field of systems neuroscience. However, existing gene delivery techniques do not permit electrophysiological characterization of cells that would establish an experimental link between physiology and genetics for understanding neuronal function. Here we demonstrate in the mouse brain in vivo that (i) neurons remain intact after 'blind' whole-cell recording, (ii) that DNA vectors can be delivered through the patch-pipette during such recordings and (iii) that these vectors drive protein expression in recorded cells for at least seven days. We illustrate the utility of this approach by recording visually-evoked synaptic responses of primary visual cortical cells while delivering DNA plasmids that permit retrograde, mono-synaptic tracing of that neuron's presynaptic inputs. By providing a biophysical profile of the cell prior to its specific genetic perturbation, this combinatorial method has captured the first synaptic and anatomical receptive field of a neuron.

Mots clés: Technical report; Neurophysiology; Synaptic Transmission; Sensory Systems; viral tracing; gene delivery

Ede A Rancz, Kevin M. Franks, Martin K. Schwarz, Bruno Pichler, Andreas T. Schaefer, et al.. Transfection via whole-cell recording in vivo: Bridging single-cell physiology, genetics and connectomics. Nature Neuroscience, 2011, ⟨10.1038/nn.2765⟩. ⟨hal-00616226⟩