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.
Behrend
Matthew R
MR
Dept. of Electrical Engineering, University of Southern California, Los Angeles, CA 90033, USA.
Ahuja
Ashish K
AK
Humayun
Mark S
MS
Weiland
James D
JD
Chow
Robert H
RH
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