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Alfonso Savastano, Chiara Adembri, Tomaso Caporossi, Stanislao Rizzo, Duccio Rossi Degli Innocenti, Francesco De Logu, Romina Nassini, Pierangelo Geppetti; Presence of transient receptor potential ankyrin1 (TRPA1) into the retinal layer. Invest. Ophthalmol. Vis. Sci. 2016;57(12):1790.
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© 2017 Association for Research in Vision and Ophthalmology.
Transient Receptor Potential (TRP) channels are essential components of biological sensors that detect environmental changes in response to a wide range of stimuli. They are permeable tmonovalent/divalent cations and contribute to changes in the cytosolic free Ca2 concentration.TRP Ankyrin 1 (TRPA1) channel subtype is a polymodal sensor expressed mainly by nociceptive neurons. TRPA1 can be activated by endogenous products generated at sites of inflammation and tissue injury, including reactive oxygen species.
Studies were conducted under the University of Florence research permit #204/2012-B.C57BL/6 mice (male, 20-25 g, age 5 weeks), littermate wild-type (Trpa1+/+) and TRPA1-deficient mice (Trpa1-/-) (25-30 g, age 5-8 weeks), generated by heterozygotes on a C57BL/6 background were used.The retinal tissue was removed, placed in 4% paraformaldehyde, and then embedded in paraffin. For immunohistochemistry, sections (4 µm), after antigen retrieval (citrate buffer solution pH 6.0) for 20 minutes at 98°C, were incubated with Ultra V Block for 5 minutes (Pierce, Thermo Scientific, Rockford, USA) following primary antibodies: TRPA1 (1:400, AVIVA System Biology, San Diego, USA) applied 1 hour at room temperature. Then, sections were incubated for 10 minutes with Primary Antibody Amplifier Quanto and AP Polymer Quanto (Pierce, Thermo Scientific, Rockford, USA). For the immunofluorescence staining, after the incubation with the primary antibody, sections were incubated with secondary antibody diluted 1:600 and counterstained with 4'6'-diamidino-2-phenylindole.
Examination of sections (n=3-5) at different levels revealed that the TRPA1 protein is present in mice retina. This finding appeared consistent in all tissue sections examined, as Immunosignals related to TRPA1 were observed in the optical nerve fibers ganglion cell layer, inner plexiform layer, inner nuclear layer and photoreceptor layer. Data confirmation was obtained with the Trpa1-/- tissue in which the signal was absent.
This is the first description of TRPA1 channel expression in the retina in the mouse. The receptor is present in different retinal layers. Its role remains to be elucidated even if it might be speculated that it might be involved in cell communication and in inflammatory processes.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
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