Direct injection into the rat optic nerve is useful for regenerative research. We demonstrate a minimally-invasive technique for direct injection into a rat optic nerve that does not involve opening the skull. Using this method, surgical complications are minimized and recovery is more rapid.
Abstract
The rat optic nerve is a useful model for stem cell regeneration research. Direct injection into the rat optic nerve allows delivery into the central nervous system in a minimally-invasive surgery without bone removal. This technique describes an approach to visualization and direct injection of the optic nerve following minor fascial dissection from the orbital ridge, using a conjunctival traction suture to gently pull the eye down and out. Representative examples of an injected optic nerve show successful injection of dyed beads.
Direct injection into the optic nerve of stem cells or other products intended to facilitate regeneration provides a convenient model compared to other means of injections into the CNS. This technique takes less time, requires less total anesthesia, avoids drilling or removing skull or bone tissue, reduces complications rates and allows for more rapid recovery following surgery.
The most critical steps in this protocol include: 1. Adequate hemostasis in the surgical field to allow clear visua…
Offenlegungen
The authors have nothing to disclose.
Acknowledgements
This study was supported by NeuralStem, Inc., and Johns Hopkins Project RESTORE.
Materials
Name of Material/ Equipment
Company
Catalog Number
Comments/Description
Lewis rat
Charles River
4
Any rat strain will work.
Anesthesia machine
Surgivet
CDS9000
CDS 9000 Small Animal Anesthesia Machine – Pole Mount
Raykova, K., Jones, M. V., Huang, H., Hoffman, P. F., Levy, M. Minimally-invasive Technique for Injection into Rat Optic Nerve. J. Vis. Exp. (99), e52249, doi:10.3791/52249 (2015).