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CSH Protoc. 2007 Jul 1;2007:pdb.prot4782. doi: 10.1101/pdb.prot4782.

Imaging retinotectal synaptic connectivity.


INTRODUCTIONThis protocol describes a method for imaging synaptic sites in individual axon or dendritic terminals in live, developing embryos. By selectively targeting expression of green fluorescent protein (GFP)-tagged pre- and post-synaptic proteins to developing neurons in vivo, it is possible to visualize synaptic sites and to correlate their distribution and dynamics with changes in the morphology of axon or dendritic arbors labeled with a red fluorescent molecule. The techniques described here may be applied to almost any neuronal circuit or neuron type, as long as the embryonic tissues are accessible for transfection and the neurons of interest are located at a depth that is within the working distance of the objective. These techniques have been optimized for Xenopus embryos, but they are applicable to other vertebrate systems such as zebrafish and chick embryos.


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