Combined Immunochemistry and Live Imaging of Fluorescent Protein Expressing Neurons in Mouse Brain
The use of transgenic mice expressing fluorescent proteins to report a specific protein or to identify specific groups of neurons in the brain is revolutionizing many different aspects of neuroscience. Here we use an example of a GFP-expressing reporter mouse from the GENSAT project that allows identification of a specific group of neurons in the mouse cortex. Live GFP detection facilitates identification of the neurons for whole-cell patch clamp electrophysiological recording to probe their functional properties. Post hoc immunohistochemistry allows specific reconstruction of the shape of the recorded neuron; this together with the detection of other co-expressed proteins helps confirm the functional identity of specific neuron types. Approaches such as these are beginning to progress the major task of untangling the complexity of a variety of brain circuits.
Key wordsGENSAT Motor cortex Whole-cell electrophysiology E-GFP reporter Layer 5
Supported by the Marsden Fund Council from Government funding, administered by the Royal Society of New Zealand. M.T. is the recipient of a Department of Physiology, University of Otago PhD scholarship.
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