Visualization of human choroidal ganglion cells with the supravital fluorescent dye 4-(4-diethylaminostyryl)-N-methylpyridium iodide.
The distribution and morphology of ganglion cell bodies and nerve fibers in supravital human choroidea is studied by staining with the fluorescent dye 4-(4-diethylaminostyryl)-N-methylpyridium iodide (4-Di-2-ASP). We have used the choroids from human donor eyes donated for cornea transplantation. The isolated tissues were rinsed in ringer solution and processed within up to 24 h after death with 3 microM of 4-Di-2-ASP in Ringer for 5-60 min at different temperatures (4-37 degrees C). After incubation, choroid whole flat mounts were observed in a fluorescence microscope. A delicate network of nerve fibers could be distinguished as soon as 5 min after immersion in 4-Di-2-ASP. In addition, labelled ganglion cell perikarya could be seen after longer incubation periods. They occurred individually or grouped in small ganglia of up to 8 cells and were linked together by various processes, which could also be traced to blood vessels. The fluorescent dye 4-Di-2-ASP provides a quick and simple method to visualize the innervation of the human choroid, including local ganglion cells. This nontoxic fluorescent technique will permit further manipulation and electrophysiological investigation of choroidal innervation.
Bergua
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A
Department of Ophthalmology, University of Erlangen-Nürnberg, Germany.
Neuhuber
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WL
Naumann
G O
GO
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002829:Choroid / Q000294:innervation*; D005456:Fluorescent Dyes; D017950:Ganglia, Sensory / Q000033:anatomy & histology*; D006801:Humans; D008856:Microscopy, Fluorescence; D008875:Middle Aged; D009412:Nerve Fibers / Q000378:metabolism; D009474:Neurons / Q000166:cytology*; D011726:Pyridinium Compounds; D014019:Tissue Donors
D005456:Fluorescent Dyes; D011726:Pyridinium Compounds; C052247:4-(4-diethylaminostyryl)-N-methylpyridinium