Cite as: Cold Spring Harb. Protoc.; 2007; doi:10.1101/pdb.prot4782

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Imaging Retinotectal Synaptic Connectivity

Susana Cohen-Cory

This protocol was adapted from "A Practical Guide: Imaging Retinotectal Synaptic Connectivity," Chapter 27, in Imaging in Neuroscience and Development (eds. Yuste and Konnerth). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2005.


INTRODUCTION

This 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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