Preparation of embryonic retinal explants to study CNS neurite growth

This protocol outlines the preparation of embryonic mouse retinal explants, which provides an effective technique to analyze neurite outgrowth in central nervous system (CNS) neurons. This validated ex vivo system, which displays limited neuronal death, is highly reproducible and particularly amenab...

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Bibliographic Details
Published in:Experimental eye research 2016-05, Vol.146, p.304-312
Main Authors: Hanea, Sonia T., Shanmugalingam, Ushananthini, Fournier, Alyson E., Smith, Patrice D.
Format: Article
Language:English
Subjects:
Analysis of Variance
Animals
Disease Models, Animal
Granulocyte-Macrophage Colony-Stimulating Factor - pharmacology
Mice
Mice, Inbred C57BL
Neurite growth
Neurites - drug effects
Neurites - pathology
Neurogenesis
Retina - drug effects
Retina - embryology
Retinal Diseases - drug therapy
Retinal explant
Retinal ganglion cell
Retinal Ganglion Cells - cytology
Tissue Culture Techniques - methods
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Summary:This protocol outlines the preparation of embryonic mouse retinal explants, which provides an effective technique to analyze neurite outgrowth in central nervous system (CNS) neurons. This validated ex vivo system, which displays limited neuronal death, is highly reproducible and particularly amenable to manipulation. Our previously published studies involving embryonic chick or adult mouse retinal explants were instrumental in the preparation of this protocol; aspects of these previous techniques were combined, adopted and optimized. This protocol thus permits more efficient analysis of neurite growth. Briefly, the retina is dissected from the embryonic mouse eye using precise techniques that take into account the small size of the embryonic eye. The approach applied ensures that the retinal ganglion cell (RGC) layer faces the adhesion substrate on coated cover slips. Neurite growth is clear, well-delineated and readily quantifiable. These retinal explants can therefore be used to examine the neurite growth effects elicited by potential therapeutic agents. •Optimized retinal explant procedure for embryonic mouse tissue.•Used to examine neurite growth in central nervous system (CNS) neurons.•Validated, amenable and highly reproducible ex vivo system.•Retinal ganglion cells (RGCs) projections can be visualized and quantified.•Application: study of growth effects induced by potential agents and genetic modifications.
ISSN:0014-4835
1096-0007
DOI:10.1016/j.exer.2016.04.004