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J.J. Pintor, J. Sanchez, M. Irazu, T. Pelaez, A. Peral; Presence of P2Y receptors in the rat retina . Invest. Ophthalmol. Vis. Sci. 2004;45(13):5378.
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Purpose: To investigate the distribution of P2Y receptors in the neural retina of the rat by immunohistochemical and immunoblotting techniques. Methods: Commercial antibodies raised for P2Y receptors were assayed in 13 days postnatal rat retinal slices. Sections of 14 mm were made using a Leyca 3050M cryostate. Immunohistochemical studies were performed starting with the following primary antibody dilutions: anti–P2Y1, 1/200; anti–P2Y2, 1/500; anti–P2Y4, 1/500; anti–P2Y6, 1/200 and anti–P2Y11, 1/1000. As secondary antibody we used goat anti–rabbit IgG–TRITC from Sigma. Protein samples (40 µg) were separated by SDS–PAGE (10% acrylamide gel) using the Bio–Rad Mini–Protein® 3–Cell System. Proteins were transferred to nitrocellulose membranes and then incubated with the mentioned antibodies at the same dilutions. Results:In the immunohistochemical analysis we found expression of P2Y2, P2Y4 and P2Y6 in the neural retina. For these receptors the highest expression were found in the outer segments of photoreceptors, followed in intensity by the outer plexiform layer and ganglion / optic nerve fibre layers. The weakest region positive to P2Y4 was the inner plexiform layer and the weakest regions labelled to the P2Y6 were the outer and inner nuclear layers and the inner plexiform layer. We did not find expression in the neural retina of the rest of P2Y receptors tested. The expression and intensity of the marker did not change when comparing areas such as central and peripheral retina. Conclusions:The pattern expression of P2Y2, P2Y4 and P2Y6 is consistent with the presence of these receptors in the plexiform layers of the retina and in the vitreous end–feet and prolongations of Müller cells (glial cells).
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