Long-lived retrograde fluorescent labeling of corticospinal neurons in the living animal.
For pathophysiological studies, it is advantageous to label specific neuronal populations in living animals. This study aimed to establish a method for stable and long-lasting fluorescent labeling of corticospinal neurons in the living animal. The two fluorescent dyes Fluoro-Red and Fluoro-Green were injected in the cervical spinal cord of anesthetized newborn rats. After a recovery period, treated rats were returned to the mother. After 24 h and 14 days, fixed brain sections revealed wide-spread fluorescence in elongated or pyramidal-shaped cell profiles in a discrete internal cortical layer, consistent with layer V pyramidal cells. Labeled neurons displayed spontaneous synaptic activity using the slice patch clamp method. These results suggest that these dyes are effective tools for pathophysiological and slice patch clamp studies focused on specific neuron groups.
Department of Basic Biological Sciences, Kyoritsu University of Pharmacy, Tokyo, Japan. suzuki-tk@kyoritsu-ph.ac.jp
Takeshi
T
Suzuki
Yoshihiro
Y
Abe
Daniel S
DS
McGehee
J Russel
JR
Keath
Hiroshi
H
Yajima
Kamal
K
Sharma
James R
JR
Brorson
D000200:Action Potentials / Q000502:physiology; D000818:Animals; D000831:Animals, Newborn; D001370:Axonal Transport / Q000502:physiology; D001562:Benzimidazoles / Q000378:metabolism; D001565:Benzoates / Q000378:metabolism; D002470:Cell Survival / Q000502:physiology; D005260:Female; D005456:Fluorescent Dyes / Q000378:metabolism*; D066298:In Vitro Techniques; D008297:Male; D008856:Microscopy, Fluorescence / Q000379:methods; D018408:Patch-Clamp Techniques / Q000379:methods; D017966:Pyramidal Cells / Q000166:cytology* / Q000502:physiology; D011712:Pyramidal Tracts / Q000166:cytology* / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D013194:Staining and Labeling / Q000379:methods*; D009435:Synaptic Transmission / Q000502:physiology; D013997:Time Factors
D001562:Benzimidazoles; D001565:Benzoates; D005456:Fluorescent Dyes; C104021:Fluoro-Green; C104022:Fluoro-Red