title: Preparation of embryonic retinal explants to study CNS neurite growth.
abstract: 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.
authors.affiliation: Department of Neuroscience, Carleton University, 1125 Colonel By Drive, Ottawa, ON, K1S 5B6, Canada. Electronic address: Patrice.Smith@carleton.ca.
authors.forename: Sonia T
authors.initials: ST
authors.lastname: Hanea
authors.affiliation: Department of Neuroscience, Carleton University, 1125 Colonel By Drive, Ottawa, ON, K1S 5B6, Canada.
authors.forename: Ushananthini
authors.initials: U
authors.lastname: Shanmugalingam
authors.affiliation: Department of Neurology and Neurosurgery, Montreal Neurological Institute, McGill University, BT-109, 3801 Rue University, Montreal, Quebec, H3A 2B4, Canada.
authors.forename: Alyson E
authors.initials: AE
authors.lastname: Fournier
authors.affiliation: Department of Neuroscience, Carleton University, 1125 Colonel By Drive, Ottawa, ON, K1S 5B6, Canada.
authors.forename: Patrice D
authors.initials: PD
authors.lastname: Smith
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D004195:Disease Models, Animal; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D051379:Mice; D008810:Mice, Inbred C57BL; D016501:Neurites / Q000187:drug effects / Q000473:pathology*; D055495:Neurogenesis; D012160:Retina / Q000187:drug effects / Q000196:embryology*; D012164:Retinal Diseases / Q000188:drug therapy; D012165:Retinal Ganglion Cells / Q000166:cytology*; D046509:Tissue Culture Techniques / Q000379:methods*
chemical_list: D016178:Granulocyte-Macrophage Colony-Stimulating Factor
keywords: Neurite growth; Retinal explant; Retinal ganglion cell