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

Margrie, Troy W ; Rancz, Ede A ; Franks, Kevin M ; Schwarz, Martin K ; Pichler, Bruno ; Schaefer, Andreas T

Nature neuroscience, 2011-04, Vol.14 (4), p.527-532 [Revista revisada por pares]

United States: Nature Publishing Group

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Transfection via whole-cell recording in vivo : bridging single-cell physiology, genetics and connectomics
Autor: Margrie, Troy W ; Rancz, Ede A ; Franks, Kevin M ; Schwarz, Martin K ; Pichler, Bruno ; Schaefer, Andreas T
Materias: Animals ; Brain - cytology ; Brain - physiology ; DNA ; Genetic aspects ; Genetic engineering ; Genetic Vectors - physiology ; Medical research ; Mice ; Mice, Inbred C57BL ; Morphology ; Neuroanatomical Tract-Tracing Techniques - methods ; Neuroanatomical Tract-Tracing Techniques - trends ; Neurons ; Neurons - cytology ; Neurons - physiology ; Neurophysiology ; Neurosciences ; Organ Culture Techniques ; Patch-Clamp Techniques - methods ; Patch-Clamp Techniques - trends ; Physiological aspects ; Physiology ; Plasmids ; Protein expression ; Proteins ; Success ; Transfection - methods ; Transfection - trends
Es parte de: Nature neuroscience, 2011-04, Vol.14 (4), p.527-532
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Author Contributions EAR, KMF, ATS, and TWM conceived experiments. KMF, ATS and TWM performed original proof-of-principle and multi-plasmid experiments. EAR collected the biocytin, GFP and viral tracing data. MKS co-designed and generated plasmids and provided virus. BP provided customized visual stimulation and compiled large-scale imaging data. EAR, KMF and TWM wrote the manuscript with input from all other authors.
These authors contributed equally
Descripción: Single-cell genetic manipulation is expected to substantially advance the field of systems neuroscience. However, existing gene delivery techniques do not allow researchers to electrophysiologically characterize cells and to thereby establish an experimental link between physiology and genetics for understanding neuronal function. In the mouse brain in vivo, we found that neurons remained intact after 'blind' whole-cell recording, that DNA vectors could be delivered through the patch-pipette during such recordings and that these vectors drove protein expression in recorded cells for at least 7 d. To illustrate the utility of this approach, we recorded visually evoked synaptic responses in primary visual cortical cells while delivering DNA plasmids that allowed retrograde, monosynaptic tracing of each neuron's presynaptic inputs. By providing a biophysical profile of a cell before its specific genetic perturbation, this combinatorial method captures the synaptic and anatomical receptive field of a neuron.
Editor: United States: Nature Publishing Group
Idioma: Inglés

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

Margrie, Troy W ; Rancz, Ede A ; Franks, Kevin M ; Schwarz, Martin K ; Pichler, Bruno ; Schaefer, Andreas T

United States: Nature Publishing Group

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