Judkewitz, B.; (2010) Optical and molecular techniques for the study of neuronal networks. Doctoral thesis , UCL (University College London).

Abstract

A central goal of systems neuroscience is to understand how brain function can be explained by the activity of intricate neuronal circuits. A major hurdle towards addressing this question has been the lack of appropriate techniques to specifically manipulate individual neurons within functioning networks and to measure their connectivity at single cell resolution. Here I present a new technique, two-photon guided single-cell electroporation, to perform genetic manipulations of individual neurons in the intact mammalian brain in vivo. I demonstrate how stable transgene expression can reliably be induced with high success rates both in single neurons as well as in spatially defined groups of neurons in the cerebral cortex of mice. Furthermore, I demonstrate how single-cell electroporation can be used in combination with retrograde viral tracing techniques to label the microcircuit impinging on a single postsynaptic target neuron. Using this approach, I present the first data of monosynaptic neuronal tracing at single-cell resolution in intact networks in vivo. These complementary techniques will provide researchers with a new approach to manipulate the function of single neurons within intact networks and to link the function of a neuronal network to the underlying circuitry.

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