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Guzel Bikbova, Toshiyuki Oshitari, Shuichi Yamamoto; Nerve fibers in rat cornea detected by direct staining by thiocholine modification of the acetylcholinesterase (AchE) technique and by anti-beta Tubulin III immunostaining. Invest. Ophthalmol. Vis. Sci. 2017;58(8):1388.
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To determine whether the corneal innervation of rats can be detected by the thiocholine modification of the acetylcholinesterase (AchE) technique and the immunohistochemical (IHC) staining of anti-beta tubulin III.
All of the procedures were performed in accordance with the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research. Ten rat corneas from 5 adult SD rats were studied. Five corneal whole mounts were stained for the cholinesterase enzyme using the Karnovsky & Roots thiocholine modification of the AchE technique. In addition, 5 corneal whole mounts were IHC with anti-beta tubulin III antibody.
AchE-positive nerves were found in the stroma of all of the corneas. The nerves entered the corneal limbus at different levels with most observed in the mid and deep stroma. The large nerve trunks were observed in the posterior lamella. Anti-beta tubulin III IHC stained mainly the corneal subbasal nerves and their associated branches.
The direct staining by the thiocholine modification of the AchE technique and ICH with anti-beta tubulin III can be used to demonstrate the rat corneal nerves in corneal whole mounts. These staining methods allowed three-dimensional observations of the distribution and spatial arrangements of the corneal nerve bundles.
This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.
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