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Updated: May 24, 2026

Conditional Genetic Transsynaptic Tracing in the Embryonic Mouse Brain

Conditional Genetic Transsynaptic Tracing in the Embryonic Mouse Brain

Published on: December 22, 2014

Transgenic strategy for identifying synaptic connections in mice by fluorescence complementation (GRASP).

Masahito Yamagata¹, Joshua R Sanes

¹Department of Molecular and Cellular Biology and Center for Brain Science, Harvard University, Cambridge MA, USA.

Frontiers in Molecular Neuroscience

|February 23, 2012

Summary

The GFP reconstitution across synaptic partners (GRASP) method now works in mice, enabling specific visualization of synapses between rod photoreceptors and other retinal neurons. This advance offers a sensitive tool for studying synaptic connections in the vertebrate brain.

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Area of Science:

Neuroscience
Molecular Biology
Genetics

Background:

The GFP reconstitution across synaptic partners (GRASP) method visualizes neuronal connections by reconstituting GFP fluorescence at synaptic sites.
GRASP has been successfully applied in invertebrates like C. elegans and D. melanogaster but not yet in vertebrates.
Existing methods for synaptic visualization in vertebrates have limitations in specificity and sensitivity.

Purpose of the Study:

To adapt and validate the GRASP method for use in mammalian systems.
To develop a transgenic strategy for selective GRASP labeling of specific neuronal populations in mice.
To enhance the sensitivity of GRASP for high-resolution synaptic imaging.

Main Methods:

Development of functional GRASP constructs for mammalian cells.

Keywords:

GFP GRASP neurexin neuroligin photoreceptor retina rod synapse

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Creation of a Cre-dependent gene switch for targeted expression of GRASP components in mice.

Generation of transgenic mice expressing Cre specifically in rod photoreceptors.

Production of antisera to amplify the GRASP signal.

Main Results:

Demonstrated successful GRASP labeling of synapses in the mouse outer plexiform layer, specifically at rod photoreceptor synapses.
Showcased the specificity of GRASP labeling, with rod synapses remaining labeled for extended periods while adjacent cone synapses were not.
Confirmed that generated antisera can amplify the GRASP signal, increasing detection sensitivity.

Conclusions:

The adapted GRASP method is effective for specific and sensitive synaptic labeling in the mouse retina.
This technique provides a valuable new tool for investigating synaptic connectivity in vertebrate neural circuits.
The enhanced sensitivity through antibody amplification allows for detailed analysis of synaptic structures.