Transection of the optic nerve in rats: studying neuronal death and survival in vivo.
Transection of the optic nerve (ON) in the adult rat, as a model of fiber tract lesion in the adult mammalian CNS, results in delayed, mainly apoptotic death of 80--90% of retinal ganglion cells (RGCs) within 14 days post-lesion. Because of good surgical accessibility of the retina and the optic nerve, the retino-tectal projection represents not only a convenient model to study the molecular mechanisms underlying neuronal death but also serves as a suitable system for investigating potential neuroprotective agents in vivo. In the present report, we provide a detailed protocol for this model including retrograde labeling of RGCs, ON lesion, assessment of the number of surviving neurons, and tissue preparation for several standard techniques like immunohistochemistry, reverse transcription--polymerase chain reaction (RT--PCR), enzyme assays and Immunoblot.
Kermer
P
P
The Burnham Institute, Program on Cell Death and Apoptosis Research, 10901 North Torrey Pines Road, San Diego, CA 92037, USA. pkermer@burnham-inst.org
Klöcker
N
N
Weishaupt
J H
JH
Bähr
M
M
D000818:Animals; D019771:Axotomy; D002452:Cell Count; D016923:Cell Death / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D005260:Female; D007150:Immunohistochemistry; D009474:Neurons / Q000502:physiology*; D018696:Neuroprotective Agents / Q000494:pharmacology; D009900:Optic Nerve / Q000166:cytology* / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013238:Stereotaxic Techniques
D018696:Neuroprotective Agents