Selective labeling of retinal ganglion cells with calcium indicators by retrograde loading in vitro.
Here we present a retrograde loading technique that makes it possible for the first time to rapidly load a calcium indicator in the majority of retinal ganglion cells (RGCs) in salamander retina, and then to observe physiological activity of these dye-loaded cells. Dextran-conjugated calcium indicator, dissolved in water, was applied to the optic nerve stump. Following dye loading, the isolated retina was mounted on a microelectrode array to demonstrate that electrical activity and calcium activity were preserved, as the retina responded to electrical stimuli.
Dept. of Electrical Engineering, University of Southern California, Los Angeles, CA 90033, USA.
Matthew R
MR
Behrend
Ashish K
AK
Ahuja
Mark S
MS
Humayun
James D
JD
Weiland
Robert H
RH
Chow
D000200:Action Potentials / Q000502:physiology; D000557:Ambystoma; D000814:Aniline Compounds; D000818:Animals; D002118:Calcium / Q000032:analysis / Q000378:metabolism*; D020013:Calcium Signaling / Q000502:physiology*; D003911:Dextrans; D004058:Diffusion; D015194:Diffusion Chambers, Culture / Q000379:methods; D004558:Electric Stimulation; D004594:Electrophysiology / Q000295:instrumentation / Q000379:methods; D005452:Fluoresceins; D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D008839:Microelectrodes; D009900:Optic Nerve / Q000166:cytology / Q000502:physiology; D009924:Organ Culture Techniques / Q000379:methods; D009930:Organic Chemicals; D051381:Rats; D020318:Rats, Long-Evans; D012165:Retinal Ganglion Cells / Q000166:cytology* / Q000378:metabolism*; D013194:Staining and Labeling / Q000379:methods*; D013997:Time Factors
C417664:Alexa594; D000814:Aniline Compounds; D003911:Dextrans; D005452:Fluoresceins; D005456:Fluorescent Dyes; D007202:Indicators and Reagents; C503942:Oregon green 488 BAPTA-1; D009930:Organic Chemicals; D002118:Calcium